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Antibiotics and the nutrient requirements of the chick Smith, David F. 1952

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S £ ft b  ANTIBIOTICS AND THE NUTRIENT REQUIREMENTS OF THE CHICK  by David F. Smith, B.S.A.  A THESIS.SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE IN AGRICULTURE i n the Department of Poultry Husbandry  We accept t h i s thesis as conforming t o the standard required from candidates f o r the degree of Master of Science i n A g r i c u l t u r e .  Members of the Department of Poultry Husbandry THE imiVERSITY OF BRITISH COLUMBIA A p r i l , 1952  ABSTRACT  ANTIBIOTICS AND  THE NUTRIENT  REQUIREMENTS 01 THE CHICK  A study has been made on the growth stimulatorye f f e c t of a n t i b i o t i c s when fed to chicks as supplements to d i f f e r e n t rations. 1280  A t o t a l of 9 b i o l o g i c a l tests employing  chicks were c a r r i e d out. Chicks fed the Connecticut r a t i o n and  modifications  thereof showed a s i g n i f i c a n t growth response to aureomycin supplementation of the rations. u t i l i z a t i o n was  The e f f i c i e n c y of feed  improved by the addition of aureomycin to  the rations i n t h i s experiment. The e f f e c t of aureomyoin has been tested i n rations i n which the protein l e v e l has beeii adjusted to 16%, 19.5%  and 22% respectively.  17.9%,  The basal rations used consisted  of ground corn and wheat as the cereal grain and soybean o i l meal and fishmeal as the protein supplements along with additional vitamins and minerals to balance the r a t i o n .  The  results showed that aureomycin did not lower the dietary requirement of the chick f o r protein. A t e s t was made on the e f f e c t of a n t i b i o t i o s on growth of chicks when fed i n rations which were d e f i c i e n t i n s p e c i f i c amino acids.  A wheat r a t i o n d e f i c i e n t i n lysine  and a corn ration d e f i c i e n t i n tryptophan have been used. The results showed that a n t i b i o t i c s did not lower the dietary requirement of the chick f o r either lysine or tryptophan. A n t i b i o t i c s were fed i n chick rations different amounts of available  carbohydrates.  containing I t was found  that a n t i b i o t i c s did not lower the dietary requirement of the chick f o r carbohydrate. A comparison of aureomycin and p e n i c i l l i n showed no difference  i n growth stimulating properties between the  two a n t i b i o t i c s under the conditions of the d i f f e r e n t experiments.  Aureomyoin supplement (aurofacj promoted the  same growth response at 0.05 and 1.0% l e v e l s . The greatest growth response to a n t i b i o t i c s was obtained when the chicks were fed a diet adequate i n available carbohydrates and protein and balanced i n amino acids.  ACIPTOWLEDGMENTS  I wish t o express my gratitude t o Professor J . B i e l y , Department of Poultry Husbandry, f o r his d i r e c t i o n i n t h i s study.  I also wish to thank Mrs.  B. E . March f o r her assistance In the s t a t i s t i c a l analysis of the data and f o r her c r i t i c i s m i n organizing the thesis material.  My appreciation i s also  extended t o Professor E. A. Lloyd, Head of the Department of, Poultry Husbandry, f o r his assistance i n proof reading the t h e s i s .  TABLE OF CONTENTS Page  I. II.  Introduction Review of L i t e r a t u r e (a) (b) (c) (d) (e) (f) (g)  III.  1  Nature of a n t i b i o t i c s E f f e c t of a n t i b i o t i c s on growth A n t i b i o t i c s and vitamins A n t i b i o t i b s and protein A n t i b i o t i c s and feed e f f i c i e n c y Action of a n t i b i o t i c s on the i n t e s t i n a l flora Comparison of a n t i b i o t i c s as growth stimulants  4 10 18 25 30 32 39  Experimental Experiment I Test No. 1  45 46  Experiment I I Test No. 2 Test No. 3 Test No. 4  61 61 69 72  Experiment I I I Test No. 5 Test No. 6 Test No. 7  77 77 85 89  Experiment IV Test No. 8 Test No. 9  92 92 96  IV.  Discussion  101  .1,  Summary  111  VI.  Bibliography  112  ANTIBIOTICS AND  THE  NUTRIENT  REQUIREMENTS OF THE  CHICK  I." INTRODUCTION  The  d i s c o v e r y t h a t a n t i b i o t i c s were of value i n  enhancing growth r a t e and  feed e f f i c i e n c y i n b r o i l e r  p r o d u c t i o n has  c r e a t e d an e n t i r e l y new  in nutrition.  Whether o r not the o l d concepts of p r o t e i n ,  energy, v i t a m i n and m i n e r a l  I t has  research  requirements f o r growing b i r d s  w i l l be a f f e c t e d thereby, has the minds o f those who  f i e l d of  become a l e a d i n g q u e s t i o n i n  are concerned with n u t r i t i o n a l problems.  been found by v a r i o u s i n v e s t i g a t o r s t h a t  i n c r e a s e s i n r a t e of growth v a r y i n g from 10-15% have been p o s s i b l e by adding a n t i b i o t i c s t o a r a t i o n . i t has  been found t h a t f e e d e f f i c i e n c y has  i n c r e a s e d i n v a r y i n g degrees with the use  Furthermore,  a l s o been of a n t i b i o t i c s .  T h i s i n t u r n has meant a f u r t h e r r e d u c t i o n i n costs t o producer.  Reports have i n d i c a t e d moreover, t h a t the  of disease has  the  incidence  been lowered and t h a t the l e s s vigorous  birds  i n the f l o c k which f o r m e r l y were unable t o compete w i t h h e a l t h i e r and  s t r o n g e r b i r d s , are now  a t t a i n b e t t e r growth.  able t o s u r v i v e  and  As a r e s u l t a g r e a t e r degree of  u n i f o r m i t y i s found i n the b i r d s i n the b r o i l e r  the  industry  -  2 -  s i n c e the a d d i t i o n o f a n t i b i o t i c s has become standard practice i n feeding. I t has been noted t h a t there  i s considerable  v a r i a t i o n i n the response o f c h i c k s t o a n t i b i o t i c s . v a r i a t i o n i s apparently  This  the r e s u l t o f d i f f e r e n c e s i n the  composition o f the r a t i o n s f e d .  An i n v e s t i g a t i o n was  undertaken t o determine the e f f e c t o f a n t i b i o t i c s on the d i e t a r y requirements o f the c h i c k f o r carbohydrates, and  protein  the amino a c i d s , l y s i n e and tryptophan. The  mode o f a c t i o n o f a n t i b i o t i c s has not y e t been  d e f i n i t e l y established.  There are a number o f ways i n which  they c o u l d promote growth.  I t has been suggested by  *Cuthbertson t h a t they may a c t by i n h i b i t i n g the m i c r o organisms t h a t d e s t r o y  o r i n a c t i v a t e growth f a c t o r s .  support has been g i v e n  t o t h i s h y p o t h e s i s by t h e o b s e r v a t i o n s  t h a t v i t a m i n B±2  removed from i n t e s t i n a l contents  c  a  n  0  6  Some  by b a c t e r i a l a c t i o n , t h a t E s c h e r i c h i a o o l i can i n a c t i v a t e v i t a m i n B 1 2 , and t h a t complete e l i m i n a t i o n o f b a c t e r i a l f l o r a may occur on f e e d i n g  aureomycin.  The decreased u t i l i z a t i o n  of v i t a m i n s by b a c t e r i a caused i n t h i s way would l e a d t o increased  a v a i l a b i l i t y t o the h o s t .  A n t i b i o t i c s could  encourage growth o f micro-organisms t h a t s y n t h e s i z e  * Nutrition".  Cuthbertson, W. F. J . 1952. J". S c i . Food A g r i c ,  growth  "Antibiotics i n  February 3; 49.  - 3 -  f a c t o r s ; f o r example, o r a l streptomycin increased population  leads t o an  o f Bacterium megatherium i n the gut  o f the mouse and t h i s organism has been shown t o produce v i t a m i n Bi2«  A n t i b i o t i c s again c o u l d a c t by e l i m i n a t i o n  o f b a c t e r i a normally, o r , under c e r t a i n c o n d i t i o n s , p r o l i f e r a t i n g i n the gut and producing t o x i n s t h a t decrease the growth p o t e n t i a l o f the organism. that Clostridium perfringens caecae o f turkeys  can be e l i m i n a t e d from the  and the faeces  are g i v e n p e n i c i l l i n  o f pigs when these animals  o r terramycin.  known t o be r e s p o n s i b l e  I t has been shown  T h i s organism i s  f o r enterotoxaemia i n sheep, and  consequently the e l i m i n a t i o n o f t h i s bacterium might w e l l i n c r e a s e the growth r a t e o f animals. I t i s h i g h l y probable t h a t no s i n g l e e f f e c t o f o r a l antibiotics i s s u f f i c i e n t to explain t h e i r action, which w i l l depend on circumstances; i t may be due t o the e l i m i n a t i o n o f t o x i c organisms or o f b a c t e r i a t h a t i n a c t i v a t e growth f a c t o r s , or even t o enhanced p r o l i f e r a t i o n o f vitamin  synthesizing bacteria.  - k -  I I . REVIEW' OF LITERATURE  (a)  Nature o f a n t i b i o t i c s . A n t i b i o t i c s have been defined, as chemical  substances produced by micro-organisms which have the capacity  t o i n h i b i t t h e growth o f and even t o d e s t r o y  b a c t e r i a and o t h e r micro-organisms. by c e r t a i n p r o p e r t i e s the o r d i n a r y  They a r e c h a r a c t e r i z e d  which d i s t i n g u i s h them s h a r p l y  from  chemical a n t i s e p t i c s and d i s i n f e c t a n t s .  A n t i b i o t i c s a r e produced p r i m a r i l y by m i c r o organisms, which are d e s i g n a t e d as " a n t a g o n i s t s "  since  they  have the p r o p e r t y o f a n t a g o n i s i n g o r i n h i b i t i n g the growth of o t h e r organisms.  Some are r e a d i l y s e c r e t e d  c u l t u r e medium, whereas others a r e r e t a i n e d microbial  i n t o the  l a r g e l y i n the  cells. One  o f the most d i s t i n g u i s h i n g p r o p e r t i e s o f  a n t i b i o t i c s i s t h e i r s e l e c t i v e a c t i o n upon b a c t e r i a and other micro-organisms.  Some are a c t i v e l a r g e l y upon  Gram-positive and o n l y upon few Gram-negative b a c t e r i a , whereas others a f f e c t a l i k e v a r i o u s these groups.  bacteria within  each o f  Some a c t upon f u n g i , whereas others do n o t .  Some a r e a l s o a c t i v e a g a i n s t have v e r y l i t t l e  rickettsia.  e f f e c t upon v i r u s e s .  Most a n t i b i o t i c s  The d i f f e r e n c e s i n  the a n t i m i c r o b i a l a c t i o n o f a n t i b i o t i c s a r e both q u a l i t i v e and  quantitive  i n nature.  One may thus speak o f a  - 5b a c t e r i o s t a t i c o r an a n t i b i o t i c spectrum, t o d e s i g n a t e the range o f s e l e c t i v e a n t i m i c r o b i a l a c t i o n o f a g i v e n upon a number o f r e p r e s e n t a t i v e  substance  b a c t e r i a and o t h e r m i c r o -  organisms. A n t i b i o t i c s v a r y g r e a t l y i n t h e i r p h y s i c a l and chemical p r o p e r t i e s .  A l a r g e number o f c h e m i c a l compounds  which possess a n t i b i o t i c p r o p e r t i e s have now been i s o l a t e d . These may e i t h e r belong t o d i s t i n c t l y d i f f e r e n t groups o f chemical compounds, o r they may show c l o s e s i m i l a r i t y t o one another.  The l a t t e r i s t r u e , f o r example, o f the  different conditions  of culture.  C e r t a i n micro-organisms more than one a n t i b i o t i c .  are capable o f producing  P e n i c i l l i n notatum and P.  chrysangenum produce not o n l y d i f f e r e n t types o f p e n i c i l l i n , but a l s o a d i f f e r e n t a n t i b i o t i c designated as p e n a t i n , p e n i c i l l i n B o r n o t a t i n ; A s p e r g i l l u s f l a v u s produces a s p e r g i l l i c a c i d and p e n i c i l l i n , described  the l a t t e r having been  under a number o f d i f f e r e n t names.  Streptomyces  g r i s e u s produces two forms o f streptomycin, a c t i d o n e , and an a n t i b i o t i c present i n the mycelium o f the organism.  The  c u l t u r e f i l t r a t e s o f P e n i c i l l i n notatum and Streptomyces g r i s e u s are a c t i v e a g a i n s t  c e r t a i n b a c t e r i a l toxins ( a n t i d o t i c  p r o p e r t i e s ) , whereas the p u r i f i e d a n t i b i o t i c s produced these organisms,  by  namely p e n i c i l l i n and s t r e p t o m y c i n do n o t  possess t h a t a c t i v i t y .  Again, some a n t i b i o t i c s are produced  by more than one organism.  P e n i c i l l i n i s formed by numerous  - 6s t r a i n s o f P e n i c i l l i n "notatum, P. ohrysogenum and A s p e r g i l l u s flavus,  and by a v a r i e t y  of other fungi.  Streptomycin i s  produced by S. g r i s e u s and S. b i k i n i e n s i s ;  streptomycin-like  substances are produced alone, o r i n mixture w i t h  streptothriein  and  of other  possibly  w i t h o t h e r a n t i b i o t i c s , by a v a r i e t y  Streptomyces s p e c i e s .  Since a n t i b i o t i c s are usually  named  a f t e r the organism p r o d u c i n g them, c o n s i d e r a b l e c o n f u s i o n has  frequently resulted  from the d u p l i c a t i o n  the  same a n t i b i o t i c , which has been s t u d i e d i n crude  p r e p a r a t i o n s i s o l a t e d from d i f f e r e n t The  o f names f o r  organisms.  e f f e c t o f the medium o r o f the s u b s t r a t e  upon t h e a n t i b a c t e r i a l a c t i v i t i e s o f d i f f e r e n t varies  greatly.  antibiotics  Some substances have no e f f e c t a t a l l  upon the a n t i b a c t e r i a l a c t i v i t i e s o f a g i v e n a n t i b i o t i c ; others may reduce c o n s i d e r a b l y such a c t i v i t y by n e u t r a l i z i n g the  a n t i b i o t i c a c t i o n by some c o n s t i t u e n t o f t h e medium such  as peptone, g l u c o s e , serum o r s a l t .  Some s u b s t r a t e s ,  blood, may adsorb the a n t i b i o t i c and completely  like  inactivate  it. The  mechanism o f a n t i b a c t e r i a l a c t i o n  a n t i b i o t i c i s d i s t i n c t from t h a t Interfere  o f another.  o f one  Some a n t i b i o t i c s  w i t h t h e growth o f micro-organisms and w i t h t h e i r  c e l l d i v i s i o n ; some i n f l u e n c e  microbial  respiration;  a f f e c t the u t i l i z a t i o n o f e s s e n t i a l m e t a b o l i t e s . a n t i b i o t i c s a r e o f t e n spoken of as p r i m a r i l y  others  Although  bacteriostatic  agents, t h e i r b a c t e r i c i d a l p r o p e r t i e s may a l s o be v e r y  - 7 pronounced; the nature o f the b a c t e r i a a f f e c t e d , t h e i r stage of growth, and the composition o f medium o r s u b s t r a t e a r e of great  importance i n t h i s c o n n e c t i o n . Different- s t r a i n s of a given b a c t e r i a l  species  vary g r e a t l y i n t h e i r s e n s i t i v i t y t o a g i v e n a n t i b i o t i c . S e n s i t i v e organisms can g r a d u a l l y  develop r e s i s t a n c e t o  c e r t a i n a n t i b i o t i c s i f allowed t o be i n contact f o r any l e n g t h o f time.  D i f f e r e n t a n t i b i o t i c s vary greatly,  however, i n the mechanism w i t h which such develops.  w i t h them  The l o s s o f r e s i s t a n c e  resistance  a l s o d i f f e r s w i t h the  a n t i b i o t i c and the s e n s i t i v e b a c t e r i a . The is f a i r l y well  b a c t e r i a l a c t i o n o f the more common a n t i b i o t i c s established.  P e n i c i l l i n i n the f r e e s t a t e , i s an organic a c i d but  i s u s u a l l y a d m i n i s t e r e d as the c a l c i u m o r sodium s a l t .  The  effectiveness  of t h i s drug depends upon s e v e r a l f a c t o r s ,  some o f which are: v a r i a t i o n i n b a c t e r i a l s t r a i n s u s c e p t i b i l i t y , number o f organisms present and t h e pH o f the medium. P e n i c i l l i n as p o i n t e d specia of fungi.  out above i s b i o s y n t h e s i z e d  by s e v e r a l  The s t r u c t u r a l formula i s : .CH3 GH3  COOH  The most commonly used form i s b e n z y l p e n i c i l l i n  (penicillin  G) where the R group has the s t r u c t u r e :  B a c t e r i a under the i n f l u e n c e of b a c t e r i c i d a l c o n c e n t r a t i o n s of p e n i c i l l i n undergo d i s t o r t i o n , and u l t i m a t e l y s i s .  The o v e r - a l l e f f e c t can be  i n terms of i n h i b i t i o n o f mononucleotide  swelling interpreted  catabolism.  I n h i b i t i o n of t h i s c a t a b o l i s m prevents the mononucleotide from l i b e r a t i n g a base that m a i n t a i n s a redox p o t e n t i a l . I f t h i s p o t e n t i a l i s lowered by p e n i c i l l i n ,  the -SH group of  amino a c i d s are p r o t e c t e d from i r r e v e r s i b l e dehydrogenation. T h i s i n t e r r u p t i o n o f c e l l metabolism causes the death of the bacterium. S i n c e the -SH groups a c t as donors f o r the "H" to. be u l t i m a t e l y accepted by atmospheric oxygen, the  dehydrogen-  a t i o n w i l l proceed more r a p i d l y as t h e r e i s more O2 a v a i l a b l e and t h e r e f o r e p e n i c i l l i n a c t s more e f f e c t i v e l y on anaerobic organisms when they are f r e e l y exposed t o oxygenation. Streptomycin i s s i m i l a r t o p e n i c i l l i n i n possessing low t o x i c i t y t o animals but d i f f e r i n g i n t h a t i t a c t s on the gram n e g a t i v e organisms and i s v e r y s l o w l y absorbed from the i n t e s t i n a l t r a c t upon o r a l a d m i n i s t r a t i o n . !  S t r e p t o m y c i n has the e m p e r i c a l formula of G  2 1 H37..39 ° 1 2  N  7*  T h i s substance i s a s a l t o f an organic  base which can be d i s s o c i a t e d and form a complex w i t h  -  ribonucleic acid.  9  -  I t i s t h i s complex which i s b e l i e v e d t o  block the metabolism o f the organism, along w i t h i n h i b i t i o n of  the p o l y n u c l e o t i d e c a t a b o l i s m r e s u l t i n g i n a l o w e r i n g of  the  redox p o t e n t i a l c a u s i n g death and l y s i s Aureomycin  of the organism.  i s a y e l l o w c r y s t a l l i z a b l e substance  b i o s y n t h e s i z e d by Streptomyces a u r e o f a c i e n s .  I t has a wide  a n t i b a c t e r i a l spectrum which i n c l u d e s a number o f gramp o s i t i v e and gram-negative organisms.  The f o r m u l a of t h i s  substance i s not known but i t i s known t o c o n t a i n 0, H, and C-2.  N,  The mechanism of the a n t i b a c t e r i a l a c t i o n remains  unsolved. B a c i t r a c i n i s a n e u t r a l compound, s o l u b l e i n water and organic s o l v e n t s .  I t i s highly active against  certain  g r a m - p o s i t i v e b a c t e r i a , has v e r y l i m i t i n g t o x i c i t y t o animals, and e x e r t s , c l i n i c a l l y ,  a marked e f f e c t i n the  treatment of i n f e c t i o n s caused by s e n s i t i v e  bacteria.  Terramycin has a c a l c u l a t e d formula f o r the f r e e , amphoteric base o f C22 2 4 - 2 6 2 O 9 . H  N  I t has been shown t h a t  a phenyl group and a c a r b o h y d r a t i c moiety i n the amphoteric t e r r a m y c i n are p r e s e n t .  Terramycin appears t o be s t a b l e i n  aqueous s o l u t i o n s i n the range o f pH3 dry, not  t o 9 at 5 ° ; and i n the  c r y s t a l l i n e s t a t e , t e r r a m y c i n and i t s h y d r o c h l o r i d e are i n a c t i v a t e d on prolonged s t o r a g e at room temperature.  Terramycin s a l t s are r e a d i l y absorbed.  The amphoteric form  - 10 -  i s h i g h l y i n s o l u b l e and i s t h e r e f o r e o n l y s l o w l y The  a n t i b a c t e r i a l s p e c t r a and the blood  produced by o r a l a d m i n i s t r a t i o n  absorbed.  concentrations  of terramycin  and aureomycin  are very s i m i l a r . Terramycin i s a c t i v e a g a i n s t  a v a r i e t y of micro-  organisms, i n c l u d i n g g r a m - p o s i t i v e and gram-negative b a c t e r i a . (b)  E f f e c t o f a n t i b i o t i c s on growth. One o f the e a r l y r e p o r t s o f the growth s t i m u l a t i n g  e f f e c t o f a n t i b i o t i c s was made by Moore and co-workers, (1) i n 1946.  I n t h e i r work they were l o o k i n g f o r a drug o r  combination o f drugs t h a t would completely i n a c t i v a t e a l l b a c t e r i a i n the i n t e s t i n a l t r a c t and provide animal so t h a t v i t a m i n  a sterile  requirements c o u l d be s t u d i e d  uncomplicated by " i n t e s t i n a l v i t a m i n s "  o r t o x i c substances.  T h e i r r e s u l t s showed t h a t s u l f a s u x i d i n e and  streptomycin  s i n g l y o r i n combination l e a d t o i n c r e a s e d growth responses i n . c h i c k s when added t o a p u r i f i e d d i e t .  Unfortunately,  these i n v e s t i g a t o r s d i d not c a r r y on f u r t h e r experiments r e s u l t i n g from t h i s growth response. I n 1949 S t o k s t a d ,  e t a l (2), r e p o r t e d  experiments  i n which i t was found t h a t crude aureomycin (product o f Streptomycin a u r e o f a c i e n s prepared from i t ,  fermentation),  or a f r a c t i o n  would produce growth response i n c h i c k s  when added t o an a l l - v e g e t a b l e d i e t c o n t a i n i n g 70% soybean o i l meal and supplemented w i t h an adequate q u a n t i t y o f v"it.B]_2.  - 11 -  More marked i n c r e a s e s w i t h aureomycin f r a c t i o n were obtained i n experiments w i t h pigs by Cunha and co-workers who  used a d i e t c o n t a i n i n g corn and peanut meal and  mented with m i n e r a l s , v i t a m i n A and D,  thiamine,  (3),  supple-  riboflavin,  n i a c i n , pantothenic a c i d , f o l i c a c i d and c h o l i n e .  In  one  experiment, p i g s r e c e i v i n g an aureomycin f e r m e n t a t i o n supplement grew at a r a t e which was  2.5  times the g a i n of  those f e d on the b a s a l r a t i o n d u r i n g a f i v e week t e s t p e r i o d . In another  experiment, C a t r o n  (4), s t u d i e d the  e f f e c t of aureomycin f e r m e n t a t i o n products and found i n c r e a s e d growth r a t e i n pigs and p r e v e n t a t i o n of diarrhea.  V a r i o u s batches  an  bloody  of supplements prepared  from  Streptomyces a u r e o f a c i e n s f e r m e n t a t i o n product were f e d to 195' p i g s .  These animals  body weight of 1.64 3.47  showed an average d a i l y g a i n i n  pounds per p i g w i t h f e e d e f f i c i e n c y of  pounds of f e e d per pounds of g a i n .  Three per cent o f  the animals were removed from the experiment due In c o n t r a s t , 50 p i g s not r e c e i v i n g t h i s showed an average d a i l t y g a i n o f 1.30 consumption of 3.74  to  enteritis.  supplementation,  pounds and  pounds per pound of g a i n .  feed  Twenty per  cent of the l a t t e r p i g s were removed from the experiment due t o e n t e r i t i s .  Carpenter  (5), o b t a i n e d s i m i l a r  results  with pigs. U s i n g t u r k e y s , McGinnis and co-workers obtained e x t r a growth response  (6),  w i t h supplements o b t a i n e d  - 12 -  from aureomycin f e r m e n t a t i o n .  A b a s a l d i e t c o n t a i n i n g 56%  of soybean o i l meal was used.  When t b i s was supplemented  w i t h l i v e r p a s t e c o n t a i n i n g Bi2» McGinnis weighing  386 grams a t f o u r weeks o f age.  obtained poults When an aureomycin  f e r m e n t a t i o n product was added a t a l e v e l o f 0.5% p o u l t s weighing  586 grams were obtained.  E x t r a growth response i n  p o u l t s w i t h aureomycin f e r m e n t a t i o n supplements has a l s o been noted by Singsen and Matterson Berg  (7).  ( 8 ) , s t u d i e d the e f f e c t on growth o f f r y e r s  by adding aureomycin f e r m e n t a t i o n product t o a h i g h energy fryer ration.  The a d d i t i o n o f 0.25% o f aureomycin  a t i o n t o a high-energy  ferment-  d i e t c o n t a i n i n g 66% maize o r wheat,  1% a l f a l f a meal, 6.8% h e r r i n g meal, 20% soybean o i l meal, 1% l i v e r m e a l and m i n e r a l supplements, gave a s i g n i f i c a n t growth response.  When the h e r r i n g meal was omitted and t h e  p r o t e i n l e v e l c o r r e c t e d by a d d i t i o n a l soybean o i l meal, the growth was the same as i n the c o n t r o l group without t h e aureomycin f e r m e n t a t i o n  product.  I n a f u r t h e r study by S t o k s t a d and Jukes ( 9 ) , 1950, i t was shown t h a t c h i c k s f e d on a corn-soybean  diet  produced  growth responses w i t h c r y s t a l l i n e aureomycin h y d r o c h l o r i d e supplementation.  S u c c i n y l s u l f a t h i a z o l e , s t r e p t o m y c i n and  3-nitro-4-hydroxyphenylarsonic but these substances  a c i d a l s o gave growth a c t i v i t y  appeared l e s s potent than aureomycin.  A study was a l s o made on d i f f e r e n t f r a c t i o n s o f aureomycin  - 13  mash.  -  Some f r a c t i o n s were o b t a i n e d from fermented mash which  was a l s o devoid of aureomycin but which s t i l l r e t a i n e d activity.  growth  Furthermore, an e x t r a c t prepared by h e a t i n g a  f r a c t i o n w i t h 0.5 N-NaOH gave a growth response i n c h i c k s although a l l the aureomycin had been destroyed*by the treatment.  Aureomycin fermented-whole mash was  a l s o heated  w i t h MaOH without d e s t r o y i n g the growth promoting a c t i v i t y , although aureomycin had been i n a c t i v a t e d .  On the b a s i s o f  a l a r g e number of experiments c r y s t a l l i n e aureomycin d i d not appear t o g i v e u n i f o r m l y as g r e a t a response as aureomycin whole mash. * W h i t e h i l l , Oleson and Hutchings ( 1 0 ) , showed t h a t the a d d i t i o n of-25mg.  aureomycin HG1 per Kg. f e e d had a  s l i g h t l y b e t t e r growth promoting e f f e c t on c h i c k s than 1000 ml. Streptomyces a u r e o f a c i e n s mash per Kg. of f e e d . response was were f e d .  a l s o produced when s t r e p t o m y c i n and  Growth  penicillin  Leuke, e t a l , ( 1 1 ) , r e p o r t e d t h a t the a d d i t i o n o f  V i t a m i n B12 and s t r e p t o m y c i n (0.05%) t o a corn-soybean d i e t f o r young pigs r e s u l t e d i n a 40% i n c r e a s e i n average  daily  gains. McGinnis, et a l , ( 1 2 ) , i n t h e i r e x p e r i m e n t a l work on t u r k e y p o u l t s showed t h a t aureomycin, s t r e p t o m y c i n , t e r r a m y c i n and p e n i c i l l i n s t i m u l a t e d the growth r a t e o f turkey poults fed a p r a c t i c a l r a t i o n ; 3-nitro-4-hydroxybenzene a r s o n i c a c i d was  also e f f e c t i v e i n stimulating  -14 -  growth.  Davies and B r i g g s  (13), showed t h a t ohick growth  s t i m u l a t i o n r e s u l t e d i n most cases, but not i n a l l ,  when a  p r a c t i c a l corn-soybean r a t i o n was supplemented w i t h e i t h e r aureomycin HC1, .procaine p e n i c i l l i n G, b a c i t r a c i n o r t e r r a m y c i n . Although much o f the work t h a t has been done shows that t h e r e i s a marked growth i n c r e a s e i n f e e d i n g  antibiotics,  there a r e r e p o r t s o f i n s t a n c e s where the a d d i t i o n o f a n t i b i o t i c s had no e f f e c t on growth rate.. Speer, et a l ,  (14),  r e p o r t e d an experiment i n which the a d d i t i o n o f aureomycin to a r a t i o n f e d t o young p i g s had no e f f e c t on growth r a t e . The  authors  e x p l a i n e d t h e l a c k o f response t o aureomycin by  the " d i s e a s e l e v e l " and t h a t h e a l t h y , w e l l - f e d p i g s , may not respond t o aureomycin i f f e d under s a n i t a r y .and d i s e a s e f r e e conditions.  S c o t t and G l i s t a (15), u s i n g c h i c k s on a  corn-soybean d i e t noted o n l y a s l i g h t , to aureomycin f o r the f i r s t  insignificant  response  few weeks w i t h ad l i b i t u m  f e e d i n g and no response when i n t a k e was l i m i t e d .  The c h i c k s  i n h i s experiment " o r i g i n a t e d rrom a w e l l f e d group o f breeding hens and were not d e p l e t e d o f A.P..F. r e s e r v e s " . Oleson, Hutchings and w h i t e h i l l (16), found t h a t  depleted  c h i c k s showed no response ,.to aureomycin i n the absence o f v i t a m i n B 1 2 i n the d i e t although  there was a marked response  t o the a n t i b i o t i c when v i t a m i n Bj.2 was present o r optimal amounts.  i n sub-optimal  B i e l y and March (17), have r e p o r t e d an  experiment i n which c h i c k s f e d a corn r a t i o n p l u s  vitamins  -  15  -  at l e v e l s above those recommended by the N.R.C. r e s u l t e d i n no i n c r e a s e i n the r a t e o f growth w i t h the a d d i t i o n of aureomycin. S c o t t et a l . (18), t e s t e d an a l l - v e g e t a b l e  ration  w i t h aureomycin (15 mg/lb.) and without aureomycin plus a number of supplements.  The t e s t r a t i o n s c o n t a i n e d ground  corn, soybean o i l meal, crude soybean o i l , DL-methionine and m i n e r a l s p l u s the known v i t a m i n s .  Corn d i s t i l l e r s ' s o l u b l e s  and d e f a t t e d l i v e r meal were the o n l y supplements which improved the b a s a l r a t i o n without aureomycin, nor d i d aureomycin alone p r o v i d e any improvement.  When aureomycin  was f e d , a l l the supplements used, except dehydrated a l f a l f a , d e f a t t e d l i v e r meal, d r i e d brewers y e a s t , and d r i e d  cereal  grass ( r y e ) , gave a d e f i n i t e growth response when added to the  basal ration.  These i n c l u d e corn d i s t i l l e r ' s s o l u b l e s ,  b u t y l f e r m e n t a t i o n s o l u b l e s ( g r a i n base), b u t y l f e r m e n t a t i o n s o l u b l e s (molasses base), a l l f e d a t the 5% l e v e l and a l s o 2% o f a grass j u i c e c o n c e n t r a t e .  These r e s u l t s support the  b e l i e f t h a t the response from an a n t i b i o t i c i s dependent, i n p a r t , on the makeup of t h e r a t i o n w i t h which i t i s f e d . 1  A number o f r e p o r t s i n the l i t e r a t u r e show t h a t  c e r t a i n i n g r e d i e n t s added t o a c h i c k r a t i o n w i l l  produce  growth s t i m u l u s whether a n t i b i o t i c s are present o r n o t . date t h i s growth f a c t o r i s u n i d e n t i f i e d .  To  H i l l (50),  r e p o r t e d t h a t the a d d i t i o n of d r i e d whey, f i s h meal o r f i s h  - 16  -  s o l u b l e s to c h i c k r a t i o n s c o n t a i n i n g soybean o i l meal as source of supplementary p r o t e i n i n c r e a s e d growth. meal was  more e f f e c t i v e i n promoting growth than  l e v e l of whey f e d .  Hill  governed to a marked .extent  the'highest  from whey or f i s h meal  by the  q u a l i t y of the  was  soybean  U s i n g a p u r i f i e d type of d i e t w i t h no source of  animal p r o t e i n , C a r l s o n  et a l , (.51), obtained  i n d i c a t e d t h a t d r i e d brewers* contained  Fish  a l s o r e p o r t e d t h a t the magnitude  of the growth response o b t a i n e d  o i l .meal.  a  evidence which  y e a s t as w e l l as f i s h meal  a h i t h e r t o u n r e c o g n i z e d growth f a c t o r .  experiments, the growth response to y e a s t  and  was  approximately equal, while  was  v e r y poor, i n d i c a t i n g t h a t t h e i r d i e t was  a f a c t o r o t h e r than B12.  In  their  f i s h meal  the response t o v i t a m i n  Wiese et a l , ( 5 2 ) ,  B^2  deficient in showed t h a t  the  a d d i t i o n of f i s h meal t o an a l l - v e g e t a b l e soybean type of c h i c k r a t i o n supplemented w i t h v i t a m i n B12  gave an a d d i t i o n a l  growth response i n c h i c k s at t e n weeks of age. (53),  Sunde et a l ,  concluded t h a t the a d d i t i o n of f i s h .solubles o r  liver  products to an a l l - v e g e t a b l e type of c h i c k r a t i o n gave i n c r e a s e i n growth over t h a t obtained supplements.  with vitamin  These r e s u l t s were obtained  type of corn-soybean r a t i o n .  Hill  an  B12  using a p r a c t i c a l  and B r a n i o n ( 5 4 ) ,  also  found t h a t the a d d i t i o n of f i s h meal o r d r i e d whey to a c h i c k r a t i o n c o n t a i n i n g adequate B12 growth response.  gave a s m a l l a d d i t i o n a l  Supplementation of the b a s a l d i e t w i t h  - 17 -  methionine  i n some o f t h e i r experiments  failed  to give a  growth i n c r e a s e comparable t o t h a t g i v e n by f i s h meal and Berg et a l , (55),  'whey.  found t h a t supplementation o f an  a l l - v e g e t a b l e type o f c h i c k d i e t w i t h v i t a m i n B12 o r w i t h 5% o f f i s h meal gave approximately the same growth response. In  c o n t r a s t , when these same supplements  combination w i t h an aureomycin c h i c k s r e c e i v i n g aureomycin  were added i n  c o n t a i n i n g f e r m e n t a t i o n product,  p l u s the f i s h meal grew  s i g n i f i c a n t l y b e t t e r than those f e d aureomycin  plus vitamin  B12.  S i m i l a r r e s u l t s have been r e p o r t e d by McGinnis  (56),  with turkey poults.  et a l ,  They observed t h a t a combination  of f i s h meal and a n t i b i o t i c gave a much g r e a t e r response e i t h e r one alone when added t o a t u r k e y s t a r t i n g composed l a r g e l y o f corn and soybean r e p o r t McGinnis  et a l , (57),  o i l meal.  than  diet I n another  s t u d i e d the e f f e c t s o f f i s h  meal, f i s h s o l u b l e s , l i v e r meal and d r i e d whey on growth o f c h i c k s and t u r k e y p o u l t s f e d d i f f e r e n t types o f r a t i o n s . They found t h a t t e r r a m y c i n with whey and l i v e r L gave g r e a t e r growth response than any of the supplements The use o f a p u r i f i e d soybean soybean  o i l meal f a i l e d  alone.  p r o t e i n as a s u b s t i t u t e f o r  t o a m p l i f y the d e f i c i e n c y o f an  u n i d e n t i f i e d growth f a c t o r f o r the c h i c k . c o n t r a s t t o the r e p o r t by M i l l  (50).  This i s i n  The a d d i t i o n o f  v i t a m i n f r e e c a s e i n o r amino a c i d s (methionine, l y s i n e and tryptophan) t o the b a s a l d i e t s f a i l e d  t o g i v e a growth  - 18 -  response.  Differences  were obtained i n response t o f i s h  s o l u b l e s t o d i f f e r e n t soybean o i l meals, however, these d i f f e r e n c e s were n o t s t a t i s t i c a l l y s i g n i f i c a n t . r e p o r t , McGinnis (58),  reported  evidence i s accumulating  f i s h meal, l i v e r L faction,- o r d r i e d whey produce growth response w i t h c h i c k s  In a l a t e r  or poults  that  greater  i n t h e presence o f  a n t i b i o t i c s than i s the ease when a n t i b i o t i c s are not added; and  conversely  t h a t a n t i b i o t i c s produce a g r e a t e r  when added t o r a t i o n s c o n t a i n i n g  these animal p r o t e i n sources  than when added t o an a l l - v e g e t a b l e type r a t i o n . one  response  At l e a s t  a d d i t i o n a l u n i d e n t i f i e d f a c t o r , present i n f i s h meal,  f i s h s o l u b l e s , c e r t a i n l i v e r f r a c t i o n s and d r i e d whey, i s r e q u i r e d f o r maximum growth o f c h i c k s and t u r k e y (c)  poults.  A n t i b i o t i c s and v i t a m i n s . Davies and B r i g g s  (13),  reported  t h a t t h e r e was no  apparent s p a r i n g a c t i o n by an a n t i b i o t i c on B12 f o r e i t h e r the c h i c k o r p o u l t when f e d a p r a c t i c a l corn-soybean r a t i o n . A t k i n s o n and Couch (25), B12  showed t h a t i n j e c t i o n o f c r y s t a l l i n e  i n t o p o u l t s f e d an a l l - v e g e t a b l e p r o t e i n (soybean o i l  meal-corn) d i e t f a i l e d  t o have any e f f e c t on growth, although  such an i n j e c t i o n d i d promote the growth o f p o u l t s f e d aureomycin.  Reynolds, e t a l , (26), u s i n g good commercial  c h i c k r a t i o n s , i n a study i n v o l v i n g 500 b i r d s from 1 day t o 4 weeks found t h a t 1/3 o f an ounce o f t e r r a m y c i n t o a t o n o f  -  19 -  feed can c u t B12 requirements by 40%. A ^ h i g h e r l e v e l i n the feed d i d net s t i m u l a t e f u r t h e r g a i n s .  I t was found t h a t  even  as t e r r a m y c i n showed a B12 s p a r i n g e f f e c t , t h e v i t a m i n a l s o showed an a n t i b i o t i c s p a r i n g e f f e c t .  They recommended  that  the optimum f o r t i f i c a t i o n o f 10 grams of t e r r a m y c i n and 6 mg. o f v i t a m i n B12 i n each t o n o f c h i c k s t a r t e r be used. Graviato-Munoz,  e t a l , (27) found t h a t  was able t o r e p l a c e v i t a m i n B^2 i synthetic diet.  n  aureomycin  the r a t m a i n t a i n e d on a  When aureomycin was allowed t o s t a n d a t  room temperature o r was a u t o c l a v e d , the a n t i b i o t i c was d e s t r o y e d .  I f such aureomycin was g i v e n  potency  iatraperitoneally  o r was f e d , i t was i n a c t i v e f o r the growth promoting e f f e c t . I t was p o s t u l a t e d t h a t aureomycin may decrease t h e number of E s c h e r i c h i a c o l i i n the i n t e s t i n a l t r a c t o f the r a t .and a l l o w the i n c r e a s e o f o t h e r b a c t e r i a such as B a c i l l u s megatherium which can produce adequate q u a n t i t i e s o f v i t a m i n B12 f o r r a t growth.  That E. c o l i has a marked  a f f i n i t y f o r v i t a m i n B12 bas a l s o been demonstrated hy Davis and M I n g i o l i ( 2 8 ) . S t o k s t a d and Jukes (29) conducted s e v e r a l experiments i n which graded l e v e l s o f B12 were f e d t o the c h i c k s i n the presence and absence o f aureomycin.  A  s p a r i n g a c t i o n o f aureomycin on v i t a m i n B j _ was noted i n 2  some experiments but n o t i n o t h e r s .  When 10 micrograms o f  v i t a m i n B12 was g i v e n i n the presence o f aureomycin, almost maximum response was o b t a i n e d w i t h the v i t a m i n , but i n t h e  - 20 -  absence o f aureomycin, 50 micrograms  of vitamin B  P  1 2  e r  k i l o g r a m of d i e t produced a markedly g r e a t e r response than 10; micrograms.  Aureomycin  o f t e n produced l i t t l e  o r no  growth  response but markedly decreased m o r t a l i t y i n c h i c k s when on the b a s a l v i t a m i n B12 d e f i c i e n t l e v e l s o f B12 were added. markedly d e f i c i e n t  d i e t or when inadequate  The c h i c k s a t h a t c h i n g were  i n B12 due t o the f a c t t h a t they were  obtained from eggs l a i d by hens which were on a d i e t i n t h i s vitamin.  deficient  A summary o f a l l experiments p e r t a i n i n g  to the e f f e c t o f aureomycin on the m o r t a l i t y o f v i t a m i n deficient  B12  c h i c k s showed t h a t i n every case the m o r t a l i t y was  lower when aureomycin was  added.  Two  explanations  p o s s i b l e t o e x p l a i n t h i s r e d u c t i o n of m o r t a l i t y . aureomycin may v i t a m i n Bi2«  are First,  reduce the requirement o f the c h i c k f o r Second, the d e f i c i e n t  c h i c k s may  have a h i g h e r  s u s c e p t i b i l i t y than normal c h i c k s t o the d e l e t e r i o u s e f f e c t of c e r t a i n microorganisms i n the i n t e s t i n a l t r a c t aureomycin may  and  decrease the number of these organisms.  An experiment by Davis and co-workers (30) carried B12  i  n  was  out t o determine the content of r a d i o a c t i v e v i t a m i n  the feces o f r a t s f e d OO^Q  v i t a m i n B12 °f aureomycin, was  and aureomycin.  The  fecal  "the r a t s f e d a soybean d i e t , w i t h or without estimated by e x t r a c t i n g aqueous homogenate  o f s t o o l specimens w i t h b u t a n o l .  A d d i t i o n o f aureomycin t o  the b a s a l d i e t produced an i n c r e a s e i n the vitamin, B12  - 21 -  content,  at a g i v e n i n t a k e o f r a d i o a c t i v e G06o«  s m a l l e r i n c r e a s e i n v i t a m i n B^g content  Considerably  c o u l d be brought  about by the a d d i t i o n o f a l a r g e r amount of CO^Q*  These  r e s u l t s , t h e r e f o r e , i n d i c a t e t h a t the growth enhancement e f f e c t o f aureomycin might be due t o the i n c r e a s e i n the p r o d u c t i o n o f v i t a m i n B]_2  and perhaps other a c c e s s o r y f a c t o r s  by the b a c t e r i a l organisms i n the i n t e s t i n a l L i i i k s w i l e r , Baumann and S n e l l , 1951 study  flora. (3D  made a  on the e f f e c t o f aureomycin on the response o f r a t s t o  v a r i o u s forms o f v i t a m i n B . D  I t was found t h a t aureomycin  i n c r e a s e d the growth of r a t s f e d low l e v e l s o f p y r i d o x i n e . T h i s i n c r e a s e was s u f f i c i e n t t o cause o v e r - a l l e r r o r s o f 10 to 100% i n a b i o a s s a y  f o r vitamin B  Q  but l a r g e l y  disappeared  when h i g h l e v e l s o f the v i t a m i n were f e d . Aureomycin caused a marked i n c r e a s e i n the growth of r a t s f e d l i m i t i n g amounts of p y r i d o x a l o r pyridoxamine.  I n the presence o f  the a n t i b i o t i c the three forms o f v i t a m i n B  D  were  approximately  equal i n growth promoting a c t i v i t y , whereas,, r a t s f e d p y r i d o x a l o r pyridoxamine i n the b a s a l r a t i o n grew l e s s than r a t s f e d an equimolar amount of p y r i d o x i n e . L i h and Baumann (32)  1951,  s e t up an experiment  u s i n g groups o f 5 weanling r a t s which were g i v e n d i e t s d e f i c i e n t i n thiamine,  appropriate  r i b o f l a v i n or pantithenic a c i d  and then graded doses o f the v i t a m i n were f e d . When a n t i b i o t i c s were added i t was found t h a t p e n i c i l l i n ,  aureomycin,  -  22  -  and s t r e p t o m y c i n s t i m u l a t e d the growth of r a t s  receiving  l i m i t i n g amounts o f thiamine, r i b o f l a v i n o r pantothenic  acid.  The a n t i b i o t i c s were most e f f e c t i v e i n d i e t s t h a t c o n t a i n e d enough v i t a m i n f o r half-maximum growth, and the growth responses  due to the a n t i b i o t i c s were approximately  equal  t o those observed when the v i t a m i n content of the d i e t doubled  i n the absence of a n t i b i o t i c s .  The  was  relative  e f f e c t i v e n e s s of the a n t i b i o t i c s v a r i e d w i t h the v i t a m i n deficiency.  In thiamine d e f i c i e n c y p e n i c i l l i n  produced  the g r e a t e s t i n c r e a s e s i n growth; aureomycin and produced  d e f i n i t e but l e s s e r i n c r e a s e s ; w h i l e  t e r r a m y c i n and 3 nitro-4-hydroxyphenyl i n a c t i v e ; some s t i m u l a t i o n was  streptomycin  Chloromycetin,  a r s o n i c a c i d were  noted w i t h  streptomycin,  Chloromycetin and t e r r a m y c i n i n the presence  o f a moderate  l e v e l o f r i b o f l a v i n ; 3-nitro-4-hydroxyphenyl  arsonic acid  and s u l f a s u x i d i n e appeared to be i n a c t i v e .  In pantothenic  a c i d d e f i c i e n c y aureomycin and s t r e p t o m y c i n were best, while p e n i c i l l i n was  l e s s a c t i v e ; the a n t i b i o t i c s a l s o s t i m u l a t e d  growth i n the absence of added pantothenic B i e l y and March (17)  acid.  i n a study of the e f f e c t  of  aureomycin and v i t a m i n s on the growth r a t e of c h i c k s , found t h a t a c h i c k r a t i o n c o n t a i n i n g v i t a m i n s a t l e v e l s above those recommended by INI.R.C. r e s u l t e d i n no i n c r e a s e i n r a t e of growth when aureomycin was d e f i c i e n t d i e t without  added.  In a f o l i c  aureomycin, i t was  at 4 weeks of age averaged  o n l y 308  acid  found t h a t c h i c k s  grams i n weight and were  -  poorly feathered.  23  -  The b i r d s r e c e i v i n g aureomycin, i n s p i t e  of a s u b o p t i m a l amount o f f o l i c a c i d i n the d i e t , averaged 410 grams and were w e l l f e a t h e r e d . t h e r e f o r e , t h a t the f e e d i n g  I t would appear,  o f aureomycin t o c h i c k s lowered  t h e i r d i e t a r y requirement f o r f o l i c  acid.  In a further  experiment, r a t i o n s t h a t were d e f i c i e n t i n r i b o f l a v i n , n i c o t i n i c a c i d , and f o l i c  a c i d r e s p e c t i v e l y , were f e d w i t h  and without c r y s t a l l i n e aureomycin.  When r i b o f l a v i n ,  n i c o t i n i c a c i d o r f o l i c a c i d were o m i t t e d from the b a s a l r a t i o n , t h e r e was a s i g n i f i c a n t decrease i n the r a t e o f growth.  The a d d i t i o n o f aureomycin t o these v i t a m i n  d e f i c i e n t rations stimulated  the growth r a t e i n each case  t o the extent t h a t the c h i c k s f e d the v i t a m i n d e f i c i e n t r a t i o n s c o n t a i n i n g aureomycin a t t a i n e d weights' s i m i l a r t o those f e d the complete b a s a l r a t i o n .  I t would appear from  these experiments t h a t d i e t a r y l e v e l s o f n i c o t i n i c a c i d , f o l i c a c i d o r r i b o f l a v i n t h a t are s u b o p t i m a l f o r maximum growth r a t e o f the c h i c k under normal c o n d i t i o n s may be adequate when aureomycin i s f e d . Sunde, e t a l , ( 3 3 ) i n a r e p o r t , p r e s e n t e d evidence to show t h a t one f u n c t i o n o f the a n t i b i o t i c s i s the s p a r i n g of v i t a m i n B3.2 and thus t h e i n d i r e c t s p a r i n g o f methionine and c h o l i n e requirements o f t h e c h i c k .  Ohicks were f e d a  g r a i n r a t i o n c o n t a i n i n g peanut meal as the p r o t e i n supplement.  The a d d i t i o n o f e i t h e r methionine, c h o l i n e o r  - 24 -  v i t a m i n B12 t o the b a s a l r a t i o n , brought a s i g n i f i c a n t s t i m u l a t i o n o f growth.  The magnitude o f the growth response  from v i t a m i n B12 a d d i t i o n s (0-200 gamma/kg.) depended upon the q u a n t i t y of the v i t a m i n added w i t h i n c r e a s e s i n growth up t o the 200 gamma p e r kg. l e v e l . response was observed  No a d d i t i o n a l growth  by combinations o f c h o l i n e , methionine,  v i t a m i n B12 and s t r e p t o m y c i n  above t h a t o b t a i n e d  (1) 30 micrograms v i t a m i n B^2 P  e r  k  S « plus  with  streptomycin,  (2) c h o l i n e and methionine o r (3) a h i g h l e v e l o f v i t a m i n Bl2Coates,  e t a l , (34) r e p o r t a study o f the e f f e c t  of a n t i b i o t i c s on the growth o f c h i c k s d e p r i v e d o f v i t a m i n s of the B complex.  They f e d p e n i c i l l i n  (2.5mg./I00g.) as  the procaine d e r i v a t i v e t o 12-16 day o l d c h i c k s f e d v i t a m i n deficient diets.  R e s u l t s showed t h a t the presence o f  a n t i b i o t i c had no e f f e c t on the degree o f d e f i c i e n c y o f r i b o f l a v i n , thiamine,  p y r i d o x i n e o r pantothenic  acid;:  It  l e s s e n e d t h a t o f b i o t i n o r f o l i c a c i d and i n c r e a s e d t h a t of n i c o t i n i c a c i d .  Although  there were i n d i c a t i o n s t h a t  the d i e t s were s l i g h t l y d e f i c i e n t i n v i t a m i n B12, the e f f e c t s o f p e n i c i l l i n were e s s e n t i a l l y the same when v i t a m i n Bj_2 was added. As p a r t o f a l a r g e r i n v e s t i g a t i o n concerning b i o c h e m i c a l responses i n c h i c k s t o a n t i b i o t i c s ,  liver  v i t a m i n A and serum c a r o t e n o i d l e v e l s have been determined  - 25 -  by Burgess, et a l , (35) diet.  The  b a s a l d i e t had 54,  ground y e l l o w corn 17,  brewers yeast  salt  gms.  5,  diet.  manganese s u l f a t e 0.3 400  oz.,  units T i t . D . ) Na  significantly  of d i e t a r y p e n i c i l l i n .  These  were Independent o f f e e d consumption and response t o the a n t i b i o t i c .  50  penicillin  elevated  increases  of body weight  L i v e r weight per lOOg. body  l e s s i n the p e n i c i l l i n - t r e a t e d group.  l i v e r V i t a m i n A,  choline  found t h a t V i t a m i n A per gram f i s h  l i v e r and t o t a l serum-carotenoids was  weight was  meat  t r i c a l c i u m phosphate  added at the r a t e of 30 mg.  I t was  by a d m i n i s t r a t i o n  f i s h meal 6,  limestone 1.5,  f i s h o i l (2400 u n i t s A,  P e n i c i l l i n was  G per kg.  the f o l l o w i n g composition ( l b . / l O G l b . ) :  a l f a l f a meal 5,  (iodized) 0.5,  c h l o r i d e 209,  i n c l u d e d i n the  s o l v e n t e x t r a c t e d soybean o i l meal  wheat m i d d l i n g s 5,  meal 4, 1.4,  when p e n i c i l l i n was  on the other hand, was  Total  i n c r e a s e d by  dietary  treatment. (d) A n t i b i o t i c s and  Protein.  B u r n s i d e , et a l , (36)  f e d r a t i o n s to p i g s i n which 41.5,  peanut meal, soybean o i l meal and  f i s h meal made up  44.5,  remainder of the r a t i o n i n  and  30%,  respectively.  The  each l o t c o n s i s t e d o f corn, bone meal 0 . 5 , s a l t 0.5%  p l u s a l l the known vitamins  shown to need.  The  limestone 0.5  which a p i g has  been  a d d i t i o n of the A.P.F. supplement t o  corn-peanut meal r a t i o n r e s u l t e d i n 2.13  and  the  times the r a t e of  - 26 o g a i n obtained on the c o n t r o l r a t i o n .  The a d d i t i o n o f the  A.P.F. supplement t o the corn-soybean  o i l meal r a t i o n  i n c r e a s e d the r a t e o f g a i n approximately  30% thus showing  t h a t AP.F supplement i s much more b e n e f i c i a l when added t o a corn r a t i o n c o n t a i n i n g peanut meal than when i t c o n t a i n s soybean o i l meal. to  The a d d i t i o n o f the A.P.F. supplement  the c o r n - f i s h meal r a t i o n was b e n e f i c i a l t o a s m a l l  extent.  The r e s u l t s showed t h a t the A.P.F. supplement  i n c r e a s e d the f e e d i n g value o f peanut meal and soybean o i l meal so t h a t these p l a n t p r o t e i n supplements were s i m i l a r i n f e e d i n g v a l u e t o the f i s h meal. Sherwood and Couch (37) determine diet  initiated  the e f f e c t o f supplementing  a study t o  a low v i t a m i n B^2  (vegetable p r o t e i n only) w i t h crude A.P.F. c o n c e n t r a t e *  on the growth o f c h i c k s up t o e i g h t t o t e n weeks o f age. The  r e s u l t s showed t h a t soybean o i l meal and cottonseed  meal o f low g o s s y p o l content may be used as the s o l e of p r o t e i n when p r o p e r l y supplemented. produced  A.P.F. concentrate  a marked i n c r e a s e i n growth. Matterson,  response  source  e t a l , ( 1 9 ) i n t h e i r study on growth  to p r a c t i c a l - t y p e chick s t a r t i n g rations with  a n t i b i o t i o s . , found t h a t a p l a n t - p r o t e i n r a t i o n i n which 2.5  *  L e d e r l e * s A.P.F. c o n c e n t r a t e - a f r a c t i o n d e r i v e d  from aureomycin f e r m e n t a t i o n .  - 27 -  per cent f i s h meal r e p l a c e d  a l i k e amount o f high  soybean o i l meal, was s u p e r i o r t o an a l l - p l a n t ration.  The g r e a t e s t  protein  protein  percentage growth response w i t h  a n t i b i o t i c s was o b t a i n e d on the poorer q u a l i t y r a t i o n s (all-plant protein).  However, the g r e a t e s t weight i s  o b t a i n e d on the b e t t e r q u a l i t y r a t i o n s ( a l l - p l a n t plus f i s h meal) supplemented w i t h a n t i b i o t i c s .  protein  The r e s u l t s  as a whole suggested t h a t the b e t t e r t h e q u a l i t y o f t h e r a t i o n with respect  t o p r o t e i n and v i t a m i n s ,  the l e s s  growth response o b t a i n e d by supplementing t h e r a t i o n w i t h antibiotics. Berg (38) has r e p o r t e d  t h a t w i t h t h e advent o f t h e  f i r s t experiments w i t h a n t i b i o t i c f e e d s t u f f s , i t appeared t h a t the growth response r e s u l t i n g w i t h t h i s m a t e r i a l when added t o an a l l soybean o i l meal p r o t e i n d i e t was so g r e a t as t o make the use o f animal p r o t e i n from f i s h meal o r meat meal no l o n g e r  a n e c e s s i t y i n high-energy b r o i l e r  feeds.  Subsequent t r i a l s have shown, however, t h a t even b e t t e r gains w i l l be made on a r a t i o n c o n t a i n i n g  f i s h meal and  a n t i b i o t i c s than w i l l be made on the a l l - v e g e t a b l e d i e t p l u s a n t i b i o t i c s and Bi2«  protein  T h i s might be a t t r i b u t e d t o  a more balanced p r o t e i n w i t h f i s h meal o r a d d i t i o n o f an unknown f a c t o r . not  As y e t t h e l e v e l o f f i s h meal t o be f e d has  been e s t a b l i s h e d s i n c e the requirement v a r i e s  according  - 28 -  to the type o f f i s h meal used o r a c c o r d i n g t o d i f f e r e n t samples the  o f soybean  o i l meal.  I n t e s t s conducted t o determine  l e v e l o f f i s h meal n e c e s s a r y f o r maximum growth w i t h  a n t i b i o t i c s , v a r i a b l e r e s u l t s i n the range o f 2 t o 7 per cent have been found. comparing In  Berg has a l s o r e p o r t e d tests.made on  r a t i o n s c o n t a i n i n g d i f f e r e n t l e v e l s of p r o t e i n .  t h r e e t r i a l s i t was found t h a t 19 p e r cent p r o t e i n  promoted as good growth as 21 p e r cent. McGinnis  (39) i n a s e r i e s o f t h r e e experiments on  turkeys f o r a p e r i o d o f f o u r weeks, s t u d i e d the e f f e c t o f p e n i c i l l i n on d i e t s c o n t a i n i n g d i f f e r e n t p r o t e i n l e v e l s .  The  r a t i o n s used were o f a p r a c t i c a l type and a l l d i e t s c o n t a i n e d the  same amount o f f i s h meal, v i t a m i n and m i n e r a l supplements.  The l e v e l o f p r o t e i n i n the v a r i o u s d i e t s was v a r i e d by changing the r e l a t i v e amounts o f soybean corn. to  o i l meal and ground  I t was found t h a t the a d d i t i o n o f p e n i c i l l i n  (5 p.p.m.)  the e x p e r i m e n t a l d i e t s c o n t a i n i n g d i f f e r e n t amounts o f  p r o t e i n d i d n o t lower the requirement of the t u r k e y p o u l t for  protein.  protein.  B e s t r e s u l t s were o b t a i n e d a t the 27% l e v e l o f  The response from p e n i c i l l i n  a t the v a r i o u s l e v e l s  was r e a s o n a b l y u n i f o r m except f o r the lowest l e v e l used.  In  t h i s p a r t i c u l a r case, a g r e a t e r response was o b t a i n e d from penicillin. M a c h l i n , e t a l , (40) s t u d i e d t h e e f f e c t o f d i e t a r y a n t i b i o t i c on p r o t e i n requirement o f growing c h i c k s when  -  f e d a corn-soybean  29  -  r a t i o n at d i f f e r e n t p r o t e i n l e v e l s .  p r o t e i n content was  The  lowered by reduced l e v e l s of c o r n and  soybean meal i n constant p r o p o r t i o n s and r e p l a c i n g them w i t h glucose (cerelose).  I t was  found i n each of t h r e e  experiments  a t f o u r and s i x weeks of age, maximal and near maximal weight was  o b t a i n e d at the 19 per cent p r o t e i n l e v e l w i t h  Without  the aureomycin  g r e a t e r weight was  aureomycin.  o b t a i n e d a t 21 o r  more per cent of p r o t e i n i n every case. The  l i t e r a t u r e reveals very l i t t l e  i n f o r m a t i o n on  the e f f e c t o f a n t i b i o t i c s i n r a t i o n s where a d e f i c i e n t amino acid occurs.  Jones  and Combs (41)  f e d p r a c t i c a l type r a t i o n s  t o c h i c k s suboptimal i n l y s i n e and tryptophan. supplementation  appeared  t o spare d i e t a r y requirement f o r  tryptophan but not f o r l y s i n e . u s i n g a corn-soybean  Aureomycin  I n another  experiment,  meal type r a t i o n , d i e t a r y p r o c a i n e  p e n i c i l l i n G d i d not spare the requirement Sherwood and Couch (37)  f o r methionine.  found by u s i n g c o t t o n s e e d meal as a  p r o t e i n supplement i t was  necessary t o add d l - l y s i n e t o  make up the amino a c i d d e f i c i e n c y even when aureomycin present i n the r a t i o n .  M a c h l i n , et a l , (42)  e f f e c t of the a d d i t i o n o f an aureomycin to a c o t t o n s e e d meal d i e t .  I t was  was  s t u d i e d the  f e r m e n t a t i o n product  found t h a t t h e r e was  s i g n i f i c a n t response when aureomycin  was  a  f e r m e n t a t i o n product  added t o the b a s a l d i e t o r t o a lysine-supplemented  - 30 -  diet.  Best  r e s u l t s were obtained when both l y s i n e and the  aureomycin f e r m e n t a t i o n  product were added.  The a d d i t i o n  of an A.P.F. supplement t o an a l l p o u l t s t a r t i n g mash c o n t a i n i n g 29% p r o t e i n and d e f i c i e n t i n l y s i n e (0.9%) has been s t u d i e d by S l i n g e r , e t a l , (43) i n r e l a t i o n t o the s e v e r i t y o f white f e a t h e r c o n d i t i o n .  When t h e d i e t  contained  29% p r o t e i n and 1.8% l y s i n e , no cases o f white f e a t h e r s appeared as a r e s u l t o f i n c l u d i n g the A.P.F. supplement. Supplementing a d i e t c o n t a i n i n g 28% p r o t e i n and 1.9% l y s i n e with p e n i c i l l i n , aureomycin o r A.P.F. r e s u l t e d i n much g r e a t e r response i n growth and f e e d e f f i c i e n c y than when such a d d i t i o n s were made t o a d i e t c o n t a i n i n g 20% p r o t e i n and 1.2% l y s i n e .  Furthermore, the a d d i t i o n o f these  m a t e r i a l s t o the l a t t e r d i e t produced a h i g h i n c i d e n c e o f white f e a t h e r s , while when added t o the former d i e t no white f e a t h e r s were noted. A.P.F. concentrates  These r e s u l t s i n d i c a t e t h a t c e r t a i n and a n t i b i o t i c s i n c r e a s e the l y s i n e  requirement o f the p o u l t f o r normal f e a t h e r pigmentation. I t i s a l s o suggested t h a t these m a t e r i a l s may i n c r e a s e the requirement o f t h i s s p e c i e s f o r other amino a c i d s . (e)  A n t i b i o t i c s and f e e d  efficiency.  Reports on the e f f e c t o f aureomycin  ferrmentation  product o r c r y s t a l l i n e aureomycin on the e f f i c i e n c y o f f e e d utilization  o f chickens,  when added t o d i e t s c o n t a i n i n g a  - 31 -  source  i f v i t a m i n B12,  (1951) (44)  are not c o n s i s t e n t .  Biely, et a l ,  showed a d e f i n i t e i n c r e a s e i n f e e d  when an aureomycin f e r m e n t a t i o n  product  efficiency  was added t o a 17  per cent p r o t e i n d i e t c o n t a i n i n g f i s h meal.  The feed  e f f i c i e n c y was improved-to the extent t h a t i t was b e t t e r than t h a t o f 21 per cent p r o t e i n a l l - v e g e t a b l e r a t i o n s c o n t a i n i n g aureomycin f e r m e n t a t i o n . (8)  Berg, e t a l ,  found no d i f f e r e n c e i n f e e d c o n v e r s i o n  fermentation  product  (1950) (15)  r a t e s when t h i s  was added t o a h i g h energy d i e t  c o n t a i n i n g f i s h meal and f e d t o c h i c k s . Glista  (1950)  n o r Groschke and Evans  N e i t h e r S c o t t and  (1950) (45)  showed an i n c r e a s e d e f f i c i e n c y when c r y s t a l l i n e aureomycin was added t o d i e t s  Bi2«  Machlin,  c o n t a i n i n g a l l the known v i t a m i n s i n c l u d i n g  e t a l , c (1951).  (40)  found t h a t aureomycin  i n c r e a s e d e f f i c i e n c y o f f e e d u t i l i z a t i o n when added t o a corn soybean d i e t c o n t a i n i n g v i t a m i n B12.  The e f f e c t o f the aureo-  mycin on f e e d e f f i c i e n c y was more pronounced w i t h i n c r e a s i n g protein  levels. A t k i n s o n and Couch,  (1951), (46)  u s i n g a soybean-  corn r a t i o n on t u r k e y poults' found an improvement i n f e e d u t i l i z a t i o n when an a n t i b i o t i c supplement was added. was v e r i f i e d by subsequent t e s t s . (13)  Davies and B r i g g s  (1951)  A  found an improved f e e d e f f i c i e n c y i n c h i c k s and p o u l t s  when t h e d i e t was supplemented w i t h a v a r i e t y The  /  This  of a n t i b i o t i c s .  r a t i o n used i n t h i s i n s t a n c e was a p r a c t i c a l  corn-  - 32 -  soybean type. I n s t u d i e s with, swine Bowland, e t a l , (1950)^ (47) r e p o r t e d a h i g h e r e f f i c i e n c y o f g a i n w i t h an aureomycin fermentation  product  than w i t h a v i t a m i n  concentrate.  Other t r i a l s  showed a g r e a t l y i n c r e a s e d r a t e and e f f i c i e n c y  of g a i n when aureomycin f e r m e n t a t i o n was f e d t o u n t h r i f t y weanling p i g s over a s i x week p e r i o d .  Rations  used i n these  experiments were both the a l l - v e g e t a b l e and mixed vegetable  p r o t e i n types.  Carpenter  animal-  (1950) (48) found s i m i l a r ;  r e s u l t s i n p i g s and noted t h a t f e e d e f f i c i e n c y was i n c r e a s e d from 6 . 4 t o 3 . 1 l b . o f f e e d / l b . g a i n by aureomycin supplementa t i o n t o an animal-vegetable  diet.  Carpenter  was a l s o able  to demonstrate an i n c r e a s e i n f e e d e f f i c i e n c y when c r y s t a l l i n e aureomycin ( 1 . 2 5 g . / l 0 0 1 b o f feed) was added t o d i e t s c o n t a i n i n g Bi2«  Beeson (1950), (49) on the o t h e r hand, u s i n g  20% p r o t e i n corn-soybean meat a l f a l f a r a t i o n s plus added vitamins was unable t o f i n d any such d i f f e r e n c e i n f e e d efficiency. (f)  A c t i o n o f a n t i b i o t i c s on the I n t e s t i n a l  flora.  In view o f the f a c t t h a t i t has been found t h a t a n t i b i o t i c s g e n e r a l l y s t i m u l a t e growth, r e s e a r c h workers have t r i e d t o determine the exact reason date, nothing  f o r t h i s response.  c o n c l u s i v e has been found although  i n v e s t i g a t i o n a l work i s i n progress.  To  considerable  Research has i n v o l v e d  -  §everal t h e o r i e s . 1.  33 -  Some o f these t h e o r i e s are as f o l l o w s :  F a c t o r s y n t h e s i s theory - i n t h i s t h e o r y i t i s  p o s t u l a t e d t h a t c h i c k growth can be g r e a t l y s t i m u l a t e d by a v i t a m i n - l i k e f a c t o r as y e t undiscovered, and t h a t one way to p r o v i d e t h i s f a c t o r i s by f e e d i n g an a n t i b i o t i c .  The  a n t i b i o t i c i s supposed t o i n h i b i t other b a c t e r i a i n the i n t e s t i n e a l l o w i n g a type t o f l o u r i s h which s y n t h e s i z e s the p o s t u l a t e d growth f a c t o r . I t i s d o u b t f u l i f such a v i t a m i n - l i k e f a c t o r i s the reason f o r added growth, s i n c e , i n o r d e r t o produce the r e s u l t s claimed by a n t i b i o t i c s , i t would have t o be much more potent than v i t a m i n B 1 2 , o r a n y t h i n g o c c u r r i n g i n distillers'  s o l u b l e s , f i s h meal, f i s h s o l u b l e s , d r i e d whey,  d r i e d brewers' y e a s t , l i v e r , 2.  or other n a t u r a l feed ingredients.  Competition t h e o r y - i n t h i s theory i t i s s t a t e d  t h a t both the animal and i t s i n t e s t i n a l microbes are competing f o r the same supply o f v i t a m i n s from the d i e t . The i n t e s t i n a l b a c t e r i a may f l o u r i s h t o such an extent t h a t they rob the animal o f v i t a m i n s and thus r e t a r d growth. Feeding an a n t i b i o t i o i s supposed t o decrease the b a c t e r i a l count, thus r e l i e v i n g the animal from c o m p e t i t i o n and p e r m i t t i n g more r a p i d growth. 3.  E n t e r o t o x i s m t h e o r y - s t a t e s t h a t a n t i b i o t i c s do  not r e a l l y s t i m u l a t e growth a t a l l ,  but r a t h e r through  their  a n t i b i o t i c a c t i o n prevent a r e t a r d a t i o n o f growth by a slow  - 34 -  p o i s o n which i s produced by harmful b a c t e r i a i n the i n t e s t i n a l tract. I n o r d e r t o a r r i v e a t a comparative b a s i s f o r study of the i n t e s t i n a l f l o r a o f the c h i c k e n , some u n d e r s t a n d i n g of the normal i n t e s t i n a l b a c t e r i a as a g a i n s t the m o d i f i e d i n t e s t i n a l bac'teria w i t h a n t i b i o t i c s , i s n e c e s s a r y . The k i n d s and numbers o f microorganisms i n the so c a l l e d normal i n t e s t i n a l f l o r a o f the c h i c k e n have been s t u d i e d by S h a p i r o and S a r l e s (59).  This  investigation  covered a p e r i o d from h a t c h i n g t o the age o f one y e a r . c h i c k s were s e l e c t e d at randon every 2 - 3 days, then every 7 - 1 4  Two  days up t o 45  days up t o the age o f 30 weeks.  A f t e r t h i s , s e l e c t i o n s were made a t i r r e g u l a r i n t e r v a l s to the age o f one y e a r .  up  The b i r d s were d e c a p i t a t e d and the  e n t i r e i n t e s t i n a l t r a c t exposed.  S p e c i f i c segments were  removed to o b t a i n i n t e s t i n a l contents f o r sampling.  The  b a c t e r i a p r e s e n t were d i v i d e d i n t o seven groups by s e l e c t i v e media.  The r e s u l t s o b t a i n e d r e v e a l e d t h a t newly hatched  c h i c k s harbour v e r y few microorganisms i n t e s t i n a l l y .  After  21 hours an i n c r e a s e o f gas producing organisms appeared. No o t h e r i n c r e a s e was noted u n t i l the b i r d s were f e d hours a f t e r h a t c h i n g ) .  (24  A r i s e i n microorganisms was noted  u n t i l 40 hours a f t e r h a t c h i n g when a peak was  reached.  The  h i g h e s t number of organisms appeared i n the caecum then a  - 35 -  p r o g r e s s i v e l y lower number i n the eolon and e s p e c i a l l y h i g h i n the c a e c a l pouches.  The c o l i f o r m were the dominant group  i n the c o l o n . B. c o l i was the predominant organism i n the c o l i f o r m group but was o f numerical contents  s i g n i f i c a n c e o n l y i n the  o f the c o l o n and c a e c a l pouches.  the enterococcus the duodenum,  I t was found t h a t  and c o l i f o r m counts were very s i m i l a r i n  i l i u m , and c o l o n .  I n the c a e c a l pouches, the  number of e n t e r o c o c e i was lower than the number o f c o l i f o r m s . L a c t o b a c i l l i were found t o be the most numerous group o f b a c t e r i a i n most areas  of the i n t e s t i n a l t r a c t .  v i t a m i n requirements o f these extensive, i t i s l i k e l y  Since the  organisms are known t o be q u i t e  these b a c t e r i a p l a y an  important  r o l e i n the v i t a m i n economy o f the h o s t . Although the l i t e r a t u r e presents  a v a r i e t y of  opinions as t o the a c t i o n o f a n t i b i o t i c s , i t i s g e n e r a l l y conceded t h a t there i s some r e l a t i o n between the i n t e s t i n a l m i c r o f l o r a and these  substances.  made (27, 28, 29, 30) suggesting  Mention has a l r e a d y been t h a t the s p a r i n g e f f e c t o f  aureomycin on v i t a m i n B12 ( t h e o r y 2) or the i n h i b i t i o n o f t o x i n producing (32)  microorganisms ( t h e o r y 3 ) .  L i h and Baumann  i n t h e i r experiment on r a t s with d i f f e r e n t  i n d i e t s d e f i c i e n t i n the B v i t a m i n s , support  antibiotics  t h e o r y 2.  They a l s o suggest t h a t s i n c e d i f f e r e n t a n t i b i o t i c s the v a r i o u s types  affect  of organisms t o d i f f e r e n t degrees, i t i s  - 36 -  not s u r p r i s i n g t h a t t h e i r growth promoting power should v a r y w i t h the v i t a m i n t h a t happens to he l i m i t i n g i n the B i e l y and March (17)  i n showing the e f f e c t of aureomycin i n  l o w e r i n g the d i e t a r y requirement support t h e o r i e s 2 and 1. that supplementing  diet.  f o r c e r t a i n B vitamins  A t k i n s o n and Gouch (25)  found  an a l l - v e g e t a b l e p r o t e i n d i e t w i t h e i g h t  c r y s t a l l i n e v i t a m i n s promoted the growth of p o u l t s i n some i n s t a n c e s but f a i l e d t o have an a p p r e c i a b l e e f f e c t i n the presence  of s t r e p t o m y c i n and v i t a m i n B±2  A.P.F. was  fed.  o  r  when aureomycin  They e x p l a i n t h i s by s u p p o r t i n g t h e o r y  2.  Speer, et a l , (14) support the i d e a of I n h i D i t i n g t o x i n producing (60) presented  b a c t e r i a ( t h e o r y 3) i n p i g s . evidence  S i e b u r t h , et a l ,  f o r t h e o r y 3 when they r e p o r t e d t h a t  t e r r a m y c i n and p e n i c i l l i n completely prevented  the growth  of C l o s t r i d i u m (probably w e l c h i i ) i n the i n t e s t i n a l t r a c t turkeys. with' p i g s .  Terramycin  of  a l s o had a s i m i l a r e f f e c t i n an experiment  T h i s organism o r one  c l o s e l y r e l a t e d to  C l o s t r i d i u m w e l c h i i has been shown to be the c a u s a t i v e agent of an entertoxemia  i n sheep.  These workers f u r t h e r hypothesized  t h a t such an organism c o u l d be producing t o x i n s i n s m a l l q u a n t i t i e s i n the i n t e s t i n e of young animals which have a growth depressing  effect.  March and B i e l y (61) found t h a t those b a c t e r i a t o be f i r s t a f f e c t e d by aureomycin f e e d i n g were the  Lactobacilli.  %  - 37 -  At the l e v e l of aureomycin c u s t o m a r i l y  added t o c h i c k  s t a r t i n g r a t i o n s w i t h A.P.F. supplements t h e r e was a s i g n i f i c a n t d e p r e s s i o n i n the number of l a c t i c a c i d b a c t e r i a present i n the f e c e s .  Increasing  the amount o f aureomycin  i n the d i e t again l e a d t o a s i g n i f i c a n t decrease i n the f e c a l l a c t i c a c i d b a c t e r i a and a s u p p r e s s i o n o f the t o t a l and  c o l i f o r m b a c t e r i a i n the f e c e s .  However, the h i g h e r  l e v e l o f aureomycin f e d d i d not s t i m u l a t e greater the  extent than the lower l e v e l .  aerobic  growth t o any  They conclude t h a t  s u p p r e s s i o n of the l a c t i c a c i d b a c t e r i a i n the i n t e s t i n e  may be a f a c t o r i n the growth s t i m u l a t i n g  e f f e c t of aureomycin  f o r the c h i c k . Dixon and Thayer (62) found t h a t f u n c t i o n a l ceca i n the c h i c k were not e s s e n t i a l f o r the growth promoting a c t i o n of a n t i b i o t i c s .  The numbers o f e n t e r o c o c c i  bacteria  i n p e n i c i l l i n f e d l o t s were lower than i n the c o n t r o l s , w h i l e the numbers i n the aureomycin f e d l o t s were h i g h e r than i n the  controls.  L a c t i c a c i d b a c t e r i a counts were c o n s i s t e n t l y  h i g h e r i n the aureomycin f e d l o t s . showed no c o n s i s t e n t  The c o l i f o r m count  t r e n d among l o t s which c o u l d  be  c o r r e l a t e d w i t h growth response. Anderson, e t a l , (63)  s t u d i e d the e f f e c t of  a n t i b i o t i c s on the i n t e s t i n a l f l o r a of chickens u s i n g containing  17, 20,  23, and 26 per cent p r o t e i n .  diets  I n the f i r s t  - 38 -  experiment an i n v e r s e r e l a t i o n s h i p e x i s t e d between pH and p r o t e i n ; p e n i c i l l i n e f f e c t e d a lowering o f pH.  Anaerobic  and m i c r o a e r o p h i l i c counts were g r e a t e r than a e r o b i c , the former decreasing, the l a t t e r remaining increasing protein.  constant  with  Both groups were i n f l u e n c e d by p e n i c i l l i n  up t o 23 per cent p r o t e i n , f o l l o w e d by a d e c i d e d w i t h 26 per cent.  decrease  A c i d u r i c were predominant over  proteolytic  types, p e n i c i l l i n e x e r t i n g the same i n f l u e n c e as on the above types. counts.  P e n i c i l l i n enhanced the c o l i i o r m and l a c t o b a c i l l i  Lowest counts were o b t a i n e d w i t h e n t e r o c o c c i .  P e n i c i l l i n reduced  these types.  When h i g h e r energy d i e t s  were f e d and aureomycin used as the a n t i b i o t i c , the r e s u l t s obtained w i t h pH, anaerobes, aerobes, s i m i l a r t o those i n the f i r s t  and e n t e r o c o c c i were  experiment.  p r o t e o l y t i c counts were reduced the l a t t e r more than the former.  A c i d u r i c and  i n the presence  of aureomycin,  G o l i f O r m counts were again .  i n c r e a s e d by a n t i b i o t i c s but-the s i g n i f i c a n c e was  reduced  at 23 per cent p r o t e i n f o l l o w e d by a s l i g h t i n c r e a s e w i t h 26 per cent.  L a c t o b a c i l l i were r e s t r i c t e d by aureomycin up  to the t h i r d week, a f t e r which they i n c r e a s e d . The  growth response  observed when s t r e p t o m y c i n  and other a n t i b i o t i c s are f e d t o turkeys was s t u d i e d by K r a t z e r , e t a l , (64)  i n r e l a t i o n t o the numbers o f y e a s t  organisms present i n the i n t e s t i n a l ' t r a c t .  Poults fed  s t r e p t o m y c i n - c o n t a i n i n g d i e t s e x h i b i t e d a 5 t o 10  fold  -  39  -  i n c r e a s e i n numbers o f i n t e s t i n a l y e a s t organisms over c o n t r o l poults.  C u l t u r e s of a y e a s t i s o l a t e d from i n t e s t i n a l contents  were f e d as supplements  (1%  of the d i e t ) t o adequate  chick  and p o u l t d i e t s and the growth r a t e s o b t a i n e d were compared w i t h those o f b i r d s f e d normal d i e t s o r supplements of antibiotics.  One s t r a i n o f y e a s t i s o l a t e d from the i n t e s t i n a l  t r a c t o f s t r e p t o m y c i n f e d p o u l t s produced a s l i g h t l y more r a p i d growth than n o r m a l . d i e t s i n some, but not i n a l l , and p o u l t f e e d i n g t r i a l s .  chick  T h i s s l i g h t growth response was  probably d i f f e r e n t from t h a t o b t a i n e d from s t r e p t o m y c i n feeding.  Day o l d c h i c k s g i v e n access f o r the f i r s t 3 days of  l i f e t o droppings o f normal o r aureomycin f e d month o l d c h i c k s grew slower than c o n t r o l c h i c k s . produced no growth d e p r e s s i o n .  A u t o c l a v e d droppings Droppings from b i r d s f e d  aureomycin produced l e s s growth d e p r e s s i o n than droppings from normal Dirds.  Aureomycin  improved growth o f the c h i c k s  f e d e i t h e r f r e s h o r a u t o c l a v e d droppings. (g)  Comparison  o f a n t i b i o t i c s as growth  stimulants.  A study of a comparison o f s e v e r a l a n t i b i o t i c s as growth s t i m u l a n t s i n p r a c t i c a l c h i c k s t a r t i n g r a t i o n s was made by Matterson, et a l , (19)  1951.  aureomycin, s t r e p t o m y c i n , p e n i c i l l i n ,  Five  antibiotics,  t e r r a m y c i n and  b a c i t r a c i n , were used as supplements t o an a l l - p l a n t r a t i o n and an a l l - p l a n t p r o t e i n r a t i o n i n which 2.5  protein per cent  -s 40  -  f i s h meal r e p l a c e d a l i k e amount o f h i g h p r o t e i n soybean o i l meal.  I t was found t h a t s t r e p t o m y c i n was the l e a s t  i n promoting  growth response  i n chicks.  t h a t although the g r e a t e s t percentage  effective  I t was a l s o noted  growth response  a n t i b i o t i c s was obtained on the poorer q u a l i t y  with  all-plant  p r o t e i n r a t i o n s , the g r e a t e s t weight was o o t a i n e d on the b e t t e r q u a l i t y , a l l - p l a n t p r o t e i n plus f i s h meal r a t i o n s . Approximately  9 grams o f a n t i b i o t i c s per t o n o f f e e d were  used so t h a t no e f f o r t was made t o determine  the l e a s t  quantity of a n t i b i o t i c necessary to obtain a given B r a n i o n and H i l l  response.  (20) found the a d d i t i o n o f  aureomycin h y d r o c h l o r i d e , p e n i c i l l i n G> potassium,  terramycin  h y d r o c h l o r i d e and s t r e p t o m y c i n s u l f a t e a t a l e v e l o f 25 m i l l i g r a m s per k i l o g r a m o f a d i e t c o n t a i n i n g 15 per cent animal p r o t e i n supplements or an a l l - p l a n t p r o t e i n  diet  supplemented w i t h c r y s t a l l i n e v i t a m i n B12 r e s u l t e d i n a growth response  i n t u r k e y p o u l t s up t o 8 weeks o f age.  The  r e l a t i v e a b i l i t y o f these a n t i b i o t i c s t o s t i m u l a t e growth was not the same from 0 t o 4 weeks as from 4 t o 8 weeks. 8 weeks o f age t e r r a m y c i n gave the g r e a t e s t growth  At  response,  f o l l o w e d , i n the case of. an animal p r o t e i n d i e t , by aureomycin, s t r e p t o m y c i n and p e n i c i l l i n and i n the case o f a p l a n t p r o t e i n d i e t by streptomycin, p e n i c i l l i n and aureomycin i n t h a t o r d e r .  However, a t f o u r weeks- o f age,  p e n i c i l l i n gave the g r e a t e s t growth response w i t h both  diets,  f o l l o w e d i n the case o f the animal p r o t e i n d i e t , hy aureomycin, t e r r a m y c i n and s t r e p t o m y c i n and i n the case of the p l a n t p r o t e i n d i e t , by t e r r a m y c i n , aureomycin and s t r e p t o m y c i n , i n t h a t order.  They suggest t h a t these r e s u l t s may  be due  to  a v a r i a t i o n .in the l e v e l of a n t i b i o t i c s r e q u i r e d a t v a r i o u s ages, a l o s s reduced  of potency upon storage i n a mixed, f e e d , a  e f f e c t w i t h continuous f e e d i n g or a s p e c i f i c i n f l u e n c e  on d e f i n i t e microorganisms  which are present a t  certain  stages but not at o t h e r s , or whose i n f l u e n c e on the host n u t r i t i o n may  be more marked a t some stages than o t h e r s .  In a r e p o r t by McGinnis  (21) data on the  critical  amounts of the v a r i o u s a n t i b i o t i c s r e q u i r e d f o r maximum growth of c h i c k s and turkeys has been presented. experiments  for aquisition  of t h i s data, he used  In h i s practical  r a t i o n s and f e d them under p r a c t i c a l c o n d i t i o n s . showed t h a t the minimum l e v e l of a p a r t i c u l a r  Results  antibiotic  i n grams per ton of f e e d necessary f o r maximum c h i c k o r p o u l t r y growth appeared antibiotics.  t o v a r y w i t h the  P r o c a i n p e n i c i l l i n appeared  different t o De more e f f e c t -  i v e than t e r r a m y c i n , aureomycin o r b a c i t r a c i n c h i c k growth.  T h i s was  2 grams per ton of f e e d . to be approximately  in  promoting  p a r t i c u l a r l y t r u e at the l e v e l of Aureomycin and t e r r a m y c i n  appeared  e q u i v a l e n t i n value f o r c h i c k growth.  -  42  -  A l e v e l o f 2 grams o f these a n t i b i o t i c s per t o n of f e e d was  as e f f e c t i v e  as a 5 gram l e v e l .  Procain p e n i c i l l i n  appeared t o be somewhat more e f f e c t i v e than t e r r a m y c i n , aureomycin o r b a c i t r a c i n  i n promoting t u r k e y p o u l t  growth.  I n a f u r t h e r r e p o r t McGinnis (22) showed the e f f e c t of removing p e n i c i l l i n from the d i e t o f p o u l t s a t d i f f e r e n t ages..  I t was  found t h a t turkeys which r e c e i v e d p e n i c i l l i n  throughout the e n t i r e  f o u r week p e r i o d had the h i g h e s t average  weights a t the end of the t r i a l . f o r the f i r s t  The f e e d i n g of p e n i c i l l i n  week of the p o u l t s ' l i f e  caused a marked  i n c r e a s e i n average weight a t f o u r weeks even though i t was removed from the d i e t when the p o u l t s were one week o f age. R e s u l t s from p r e v i o u s experiments w i t h both c h i c k s and p o u l t s i n d i c a t e d t h a t the a n t i b i o t i c supplement  c o u l d be removed  from the d i e t when they were e i g h t weeks o f age without materially affecting  average weight a t twenty weeks of age.  I n a r e p o r t on h i g h versus low l e v e l a n t i b i o t i c f e e d i n g , McGinnis (22) f e d a p r a c t i c a l t u r k e y r a t i o n t o which was the  added 50 p.p.m. of t h r e e a n t i b i o t i c s .  Increasing  l e v e l of diamine p e n i c i l l i n from 2 t o 50 p.p.m. d i d not  improve growth. supplement.  A s i m i l a r f i n d i n g was made w i t h the  bacitracin  I n c o n t r a s t , i n c r e a s i n g the l e v e l o f t e r r a m y c i n  from 5 t o 50 p.p.m. d i d appear t o g i v e an improved growth response.  A comparison o f pure t e r r a m y c i n h y d r o c h l o r i d e  - 43  and  t e r r a m y c i n c o n t a i n e d i n the  -  a n t i b i o t i c f e e d supplement  at a l e v e l of 50 p.p.m. showed these two approximately the  same growth.  As  was  p o i n t e d out  i n a r e p o r t by McGinnis, t e r r a m y c i n and graded l e v e l s produced no  antibiotics tenfold responses obtained, may  the  aureomycin f e d  three  materially  different  a l t e r the  results  o b t a i n e d by f e e d i n g h i g h  be e n t i r e l y  F r i t z (23)  different.  f e d graded l e v e l s of  a n t i b i o t i c s i n p r a c t i c a l startixig rations  to c h i c k s  e f f o r t to determine the minimum  penicillin,  and  several and  quantities  which would a f f o r d good growth s t i m u l a t i o n . bacitracin,  growth  under f i e l d c o n d i t i o n s where desease  l e v e l s of a n t i b i o t i c s may  p o u l t s i n an  at  Under l a b o r a t o r y c o n d i t i o n s ,  l e v e l of the  d i d not  be encountered, the  Kramke and  previously  e f f e c t i n growth when these  a n t i b i o t i c s were i n c r e a s e d . therefore, increasing  m a t e r i a l s to g i v e  Aureomycin,  t e r r a m y c i n a l l gave  essentially  optimum growth s t i m u l a t i o n when used at the  r a t e of 10 grams  per  penicillin  ton  of feed.  At  lower l e v e l s of i n t a k e  proved to be the most e f f e c t i v e , g i v i n g  practically  good r e s u l t s w i t h 5 grams of a n t i b i o t i c per at comparable l e v e l s was  much l e s s e f f e c t i v e .  i v e requirements were s i m i l a r f o r c h i c k s and Maximum percentage gains due about 4 weeks of  age.  ton. The  as  Streptomycin quantitat-  poults.  to a n t i b i o t i c s were observed  at  - 44 -  Reynolds,, et a l , (24) made a study o f the comparison of t e r r a m y c i n  and p e n i c i l l i n a t v a r i o u s l e v e l s t o b r o i l e r s .  They f e d a complete p r a c t i c a l type  b r o i l e r ration containing  5 per cent f i s h meal, 2.5 per cent d r i e d whey and no BQ_2 supplement.  The d i e t s were supplemented with 2 grams  penicillin,  5 grams p e n i c i l l i n ,  2 grams t e r r a m y c i n ,  terramycin,  10 grams t e r r a m y c i n  and a comhination of 2.5  grams t e r r a m y c i n ively.  and 2.5 grams p e n i c i l l i n ,  R e s u l t s showed t h a t t e r r a m y c i n  g a i n i n body weight and most e f f i c i e n t  5 grams  per t o n r e s p e c t -  produced the g r e a t e s t utilization  of feed.  No s t a t i s t i c a l l y s i g n i f i c a n t d i f f e r e n c e between groups were revealed.  Penicillin  and t e r r a m y c i n were  approximately  e q u i v a l e n t gram f o r gram i n t h e i r growth promoting p r o p e r t i e s and two grams/ton o f e i t h e r a n t i b i o t i c appeared t o be near the optimum l e v e l  f o r growth promoting p r o p e r t i e s .  No  s t a t i s t i c a l l y s i g n i f i c a n t d i f f e r e n c e s between groups were revealed.  -  45  -  I I I * EXPERIMENTAL  Experiment  1  i  Introduction The f i r s t  study was undertaken t o compare the  e f f e c t of a n t i b i o t i c s on c h i c k growth when f e d i n t h r e e different rations.  The f i r s t r a t i o n was  the s t a n d a r d  C o n n e c t i c u t r a t i o n c o n t a i n i n g ground y e l l o w corn as the c e r e a l g r a i n and f i s h m e a l , meat meal, soybean o i l meal and l i v e r meal as the p r o t e i n supplements.  This r a t i o n i s  noted f o r i t s h i g h energy, low f i b r e content. r a t i o n was  The second  an a l l - p l a n t p r o t e i n r a t i o n c o n t a i n i n g ground  ground y e l l o w corn as the c e r e a l g r a i n and soybean o i l meal as the p r o t e i n supplement.  The t h i r d r a t i o n a l s o contained  ground y e l l o w corn as the c e r e a l g r a i n but f i s h m e a l as the p r o t e i n supplement. of  F u r t h e r i n g r e d i e n t s were added t o each  the t h r e e r a t i o n s i n order t o balance  them f o r v i t a m i n s  and m i n e r a l s . Supplements  composed of v i t a m i n s and m i n e r a l s .  were added t o each o f the above b a s a l r a t i o n s i n o r d e r to  determine whether any a d d i t i o n a l growth response would  result.  A n t i b i o t i c s were added t o these  supplemented  r a t i o n s t o study t h e i r response on growth of c h i c k s as  - 46 -  Gompared t o the unsupplemented  rations with a n t i b i o t i c s  T E S T NO.  1  Purpose The purpose o f t h i s experiment was the  to determine  e f f e c t on growth of c h i c k s o f adding a n t i b i o t i c s t o  three d i f f e r e n t  p r a c t i c a l r a t i o n s and t o make a comparison  of the growth response t o each r a t i o n .  A l s o , t o study the  e f f e c t o f adding v i t a m i n s and m i n e r a l s t o these r a t i o n s  and  to determine i f any a d d i t i o n a l response c o u l d be g a i n e d when a n t i b i o t i c s were added w i t h these supplements. Methods and Procedure Day o l d , male, New Hampshire banded and d i s t r i b u t e d  c h i c k s were wing-  a t random i n twelve compartments o f  e l e c t r i c a l l y heated, Jamesway b a t t e r y brooders. c h i c k s were d i s t r i b u t e d  t o each compartment.  The  Twenty chicks  were h e l d i n these b a t t e r i e s u n t i l the end of the experiment at f i v e weeks.  The temperature was  a d j u s t e d as the b i r d s  grew o l d e r . The b i r d s were f e d mash and water ad l i b . g r i t was  f e d t o them weekly.  Granite  The f e e d on hand was r e c o r d e d  at the end o f the t h i r d , f o u r t h and f i f t h weeks f o r the purpose o f c a l c u l a t i n g  feed e f f i c i e n c y .  f e e d e f f i c i e n c y i s presented i n Table 11.  A r e c o r d of the  - 47 -  The b i r d s were weighed a t the end o f each week and a s t a t i s t i c a l a n a l y s i s made on the weights The weights F i g u r e s 1,  a t f i v e weeks.  are p r e s e n t e d i n Table 11 and g r a p h i c a l l y i n 2 and 3.  The a n a l y s i s i s presented i n Table IV.  The o r i g i n a l C o n n e c t i c u t r a t i o n was used as the first  test ration.  D r i e d brewers' yeast, however, was used r  t o r e p l a c e the b u t y l f e r m e n t a t i o n s o l u b l e s and i n a d d i t i o n G.2 gms. o f r i b o f l a v i n were added t o assure a s u f f i c i e n t amount o f t h i s v i t a m i n .  The c h o l i n e c h l o r i d e (25%)  was  i n c r e a s e d from 32 gms. t o 200 gms. ( 0 . 4 4 l b . ) t o be assured of a good m e t h y l a t i n g donor. I n the soybean r a t i o n 5 9 . 8 l b . o f c o r n and 34 l b . of soybean were used.  I n view o f the f a c t t h a t i t was an  a l l - v e g e t a b l e r a t i o n one pound o f limestone and 2 l b . o f bonemeal were added t o balance the m i n e r a l s .  This r a t i o n  needed an a d d i t i o n a l s u p p l y o f the B complex v i t a m i n s . supply these 2 l b . o f d i s t i l l e r s *  s o l u b l e s , along w i t h s m a l l  q u a n t i t i e s o f c h o l i n e c h l o r i d e (25%), n i a c i n , Ca r i b o f l a v i n and v i t a m i n Bx2 were added. i n methionine  To  pantothenate,  S i n c e soybean i s low  0 . 1 l b . were added t o make up the d e f i c i e n c y .  The f i s h m e a l r a t i o n c o n t a i n e d 74.8 l b . o f corn and 2 1 . 0 l b . o f f i s h m e a l .  One pound o f limestone was added  to balance the c a l c i u m l e v e l .  The same B-complex v i t a m i n s  were added as i n the soybean r a t i o n except t h a t v i t a m i n B12 was excluded s i n c e the f i s h m e a l s u p p l i e d an adequate amount.  - 48 Methionine was  a l s o excluded s i n c e f i s h m e a l s u p p l i e s enough  of t h i s amino a c i d . The product was  supplementary v a l u e o f an a n t i b i o t i c  t e s t e d by supplementing the three b a s a l r a t i o n s  w i t h 0.3%  of the p r e p a r a t i o n i . e . r a t i o n #2  r a t i o n #6  (Soybean) and r a t i o n #10  p r e p a r a t i o n was and m i n e r a l s  (Fishmeal).  supplementary v a l u e tested.  The  more b a s a l r a t i o n s the v i t a m i n s  (Fishmeal).  of seven v i t a m i n s  v i t a m i n and m i n e r a l  (Soybean) and r a t i o n #11  vitamins  #4  (Soybean) and r a t i o n #12  added t o the three b a s a l s i . e . r a t i o n #3  r a t i o n #7  The  and added t o the b a s a l s i . e . r a t i o n  The m i n e r a l s was  (Connecticut),  a l s o combined with a supplement of  ( C o n n e c t i c u t ) , r a t i o n #8  was  fermentation  supplement (Connecticut),  (Fishmeal).  and m i n e r a l s  and s i x  To  three  were added w i t h  the a n t i b i o t i c p r e p a r a t i o n and f e d i n r a t i o n s as  described  i n the preceding  thiamin  hydrochloride, benzoic  F i f t y m i l l i g r a m s of  5 grams of i n o s i t o l , 4.54  a c i d , 22.7  menadione and  paragraph.  136  grams of para amino  m i l l i g r a m s of f o l i c a c i d , 22.7 m i l l i g r a m s of a l p h a - t o c o p h e r o l  per hundred pounds of f e e d were the v i t a m i n s grams of f e r r i c c i t r a t e ,  500  milligrams  of copper  of  ( v i t a m i n E)  added.  25 m i l l i g r a m s of n i c k e l carbonate, 50 m i l l i g r a m s a c e t a t e , 25 m i l l i g r a m s of hydrated  mgs.  Five  sulphate, of c o b a l t  aluminium sulphate  and  25 m i l l i g r a m s o f z i n c a c e t a t e per hundred pounds of f e e d  - 49 were the m i n e r a l s added. The composition o f the b a s a l r a t i o n s supplements t o the b a s a l r a t i o n s  and the 1.  are shown i n Table  Results At  f i v e weeks o f age the c h i c k s r e c e i v i n g the  C o n n e c t i c u t b a s a l r a t i o n weighed an average o f 461 grams or 27 grams l e s s than the average weight o f the b i r d s soybean b a s a l r a t i o n  (#5).  The b i r d s  on the  on the f i s h m e a l b a s a l  weighed an average o f 426 grams o r 35 grams l e s s than those on the C o n n e c t i c u t r a t i o n and 52 grams l e s s than those on the  soybean r a t i o n .  These r e s u l t s i n d i c a t e  response t o growth w i t h the soybean  the g r e a t e s t  ration.  When' the aureomycin f e r m e n t a t i o n product was added t o the C o n n e c t i c u t b a s a l r a t i o n weight o f the b i r d s  (#2)  the average  a t f i v e weeks was 560 grams.  weights were 99 grams more than the b a s a l r a t i o n (#2)  soybean r a t i o n w i t h the aureomycin supplement  These (#1).  The  produced  an average weight o f 525 grams o r 37 grams more than the basal ration  (#5)  and 35 grams l e s s than the C o n n e c t i c u t  r a t i o n w i t h the aureomycin supplement  (#2).  r a t i o n plus the aureomycin supplement  (#10)  weighing an average o f 473  The f i s h m e a l produced c h i c k s  grams o r 53 grams more than the  average of the c h i c k s on the b a s a l r a t i o n  (#9).  The  average weights from t h i s supplemented r a t i o n were 87 grams  - 50 -  l e s s than the C o n n e c t i c u t supplemented r a t i o n (#2) and 52 grams l e s s than the soybean supplemented r a t i o n (#6). results indicate the  These  that t h e g r e a t e s t response was gained from  aureomycin supplementation t o t h e C o n n e c t i c u t b a s a l . The v i t a m i n and m i n e r a l mixture when added t o the  C o n n e c t i c u t b a s a l r a t i o n (#3) r e s u l t e d  i n average weights  of 474 grams o r 13 grams more than the b a s a l r a t i o n  (#1).  When t h i s mixture was added t o the soybean b a s a l r a t i o n the  (#7)  average weights were 452 grams o r 36 grams l e s s than the  b a s a l r a t i o n (#5).  When added t o t h e f i s h m e a l b a s a l  ration  (#11), the average weights were 471 grams o r 45 grams more than the b a s a l r a t i o n (#9).  These r e s u l t s  indicate,  t h e r e f o r e , that the g r e a t e s t response from the v i t a m i n and m i n e r a l supplement was t o be g a i n e d when added t o the f i s h m e a l basal ration. The v i t a m i n and m i n e r a l supplement when added t o the C o n n e c t i c u t b a s a l r e s u l t e d  and aureomycin i n an average  weight o f 551 grams o r 9 grams l e s s than the aureomycin supplement r a t i o n (#2) and 77 grams more than the v i t a m i n and m i n e r a l supplemented r a t i o n (#3).  T h i s combination  added t o the soybean b a s a l (#8) r e s u l t e d  i n an. average  weight of 523 grams or 2 grams l e s s than the aureomycin supplemented b a s a l (#6) and 71 grams more than the v i t a m i n and m i n e r a l supplemented r a t i o n (#7).  The combination  - 51 -  added t o the f i s h m e a l b a s a l r a t i o n (#12) r e s u l t e d i n an average weight o f 530 grams o r 57 grams more than the aureomycin supplemented r a t i o n (#10) and 59 grams more the v i t a m i n and m i n e r a l supplemented r a t i o n (#11).  than  These  r e s u l t s i n d i c a t e t h a t there was no advantage i n f e e d i n g a supplement of v i t a m i n s , m i n e r a l s and aureomycin  fermentation  product over the a d d i t i o n o f aureomycin f e r m e n t a t i o n alone.  product  Some advantage was gained when the v i t a m i n and  m i n e r a l supplement and aureomycin was added t o the f i s h m e a l ration.  The f a c t t h a t v i t a m i n s and m i n e r a l s added t o the  f i s h m e a l r a t i o n a l s o gave a response  t o growth i n d i c a t e d  t h a t f i s h m e a l must be l i m i t i n g i n some f a c t o r t h a t i s s u p p l i e d by these a d d i t i o n s . The  s t a t i s t i c a l a n a l y s i s on the f i f t h week average  weights i n d i c a t e d a s i g n i f i c a n c e a t the %  l e v e l f o r the  i n t e r a c t i o n of r a t i o n s , a n t i b i o t i c s , and v i t a m i n s and minerals.  S i n c e t h i s f i g u r e r e p r e s e n t e d the v a r i a t i o n , i t  was used as the t e s t i n g e r r o r .  The r e s u l t s then showed t h a t  the a d d i t i o n of a n t i b i o t i c s gave a t e s t o f s i g n i f i c a n c e . The  r e s u l t s from the f e e d u t i l i z a t i o n data  indicate  t h a t the C o n n e c t i c u t and f i s h m e a l b a s a l r a t i o n s are the most efficient.  The a d d i t i o n of the aureomycin f e r m e n t a t i o n  t o the b a s a l s produced an improved e f f i c i e n c y i n a l l r a t i o n s (#2,  4, 6, 8, 10, 12) where i t was used.  On the other hand ,  vitamins and m i n e r a l s when used as a supplement (#3, 4, 7, 8, 11, 12) had l i t t l e  e f f e c t on f e e d u t i l i z a t i o n .  Conclusions 1.  Aureomycin f e r m e n t a t i o n product, when added t o the C o n n e c t i c u t , soybean and f i s h m e a l b a s a l r a t i o n s r e s u l t e d i n a s i g n i f i c a n t i n c r e a s e i n growth i n chicks.  The h i g h e s t i n c r e a s e was obtained w i t h  the C o n n e c t i c u t 2.  ration.  The a d d i t i o n o f v i t a m i n s and m i n e r a l s t o these b a s a l r a t i o n s produced'no s i g n i f i c a n t weight i n c r e a s e s i n the C o n n e c t i c u t and soybean r a t i o n s . Some i n c r e a s e was noted when these supplements were added t o the f i s h m e a l b a s a l r a t i o n .  3.  The v i t a m i n and m i n e r a l supplement and aureomycin f e r m e n t a t i o n product r e s u l t e d i n no i n c r e a s e i n weight over the supplementation  w i t h aureomycin  f e r m e n t a t i o n alone when added t o the C o n n e c t i c u t and soybean b a s a l r a t i o n s .  Again, however, an  i n c r e a s e i n weight was noted when t h i s  combination  was added t o t h e f i s h m e a l b a s a l r a t i o n . 4.  The C o n n e c t i c u t and f i s h m e a l r a t i o n s were most e f f i c i e n t l y u t i l i z e d by the c h i c k s .  The aureomycin  f e r m e n t a t i o n supplement improved the e f f i c i e n c y i n a l l r a t i o n s but t h i s was not f u r t h e r improved  - 53 -  when added w i t h the v i t a m i n and m i n e r a l supplement. The v i t a m i n and m i n e r a l supplement alone d i d not improve the f e e d  efficiency.  - 54 -  Table  Composition  1  of B a s a l  Rations  Basal Basal R a t i o n #1 R a t i o n #4 Connecticut Soybean  Basal R a t i o n #8 Fishmeal  lb./1001b. lb./1001b. lb./1001b.  Ground y e l l o w corn Fishmeal Meatmeal Soybean o i l meal ( s o l v e n t ) Livermeal Ground Limestone Bonemeal A l f a l f a l e a f meal D r i e d Brewers' Y e a s t Dried D i s t i l l e r s ' Solubles Iodized S a l t Manganese Sulphate Feeding O i l (2250A; 300D) C h o l i n e c h l o r i d e {25%) Nicotinic acid Calcium pantothenate Riboflavin dl-Methionine v i t a m i n Bl2  R a t i o n #2  "  "  #3 #4 #6 #7 #8 #10  "  #12  "  #H  - f1  -  f  1  #,  X  #5 #5 #5 #9 - #9 - #9  69.0 8.0 8.0 8.0 3.0  -.  1.0 2.0 -  0.5 0.025 0.25 0.44 0.9 gm. 0.5 gm. 0.2 gm. -  -  59.8  74.8 21.0  -  -  - •  34.0  -  1.0 2.0  1.0  -  -  -  -  —  2.0 0.5 .0.025 0.25 0.44 0.9 gm. 0.5 gm. 0.2 gm. 0.1 l b . 1.0 mg.  2.0 0.5 0.025 0.25 0.44 •0.9 gm. 0.5 gm. 0.2 gm. -  -  plus a n t i b i o t i c * tr v i t a m i n and m i n e r a l . tt a n t i b i o t i c and v i t a m i n and m i n e r a l . tt antibiotic. n v i t a m i n and m i n e r a l . ft a n t i b i o t i c and v i t a m i n and m i n e r a l . tt antibiotic. tt v i t a m i n and m i n e r a l . tt a n t i b i o t i c and v i t a m i n and m i n e r a l .  L e d e r l e ' s A.P.F. 0.3%  - an aureomycin f e r m e n t a t i o n  preparation.  - 55 11  Table  Weekly Average Weights  , Ration No.  1 2  • 3 4  Connecticut  Ration  • Ration  1  2  Basal Antibiotic Vitamins & M i n e r a l s Vitamins, M i n e r a l s & Antibiotic  75 g.  Weeks  80  75 83  3  142 g . 162 145 163  4  5  218 g • 330 g. 461 g  267 239 267  396 357 399  560 474 551  4  5  Soybean R a t i o n Ration •No.  5 6 7 8  1  Ration Basal Antibiotic Vitamins & M i n e r a l s Vitamins, Minerals & Antibiotic  72 g. 80  70 79  Fishmeal Ration No. 9 10 11 12  Ration Basal Antibiotic Vitamins & M i n e r a l s Vitamins, Minerals & Antibiotic  2  Weeks  3  140 g. 234 g . 344 g. 248 150 380 228 133 361 248 149 374  488 g 525 452 523 .  Ration 1  73 g. 79 71 78  2  Weeks 3  4  5  129 g. 206 g. 315 g. 426 g. 150 232 350 473 130 207 348 471 157 258 384 530  - 56 -  T a b l e 111 Feed E f f i c i e n c y Connecticut Ration Ration No. 1 2 3 4  . Ration  Weeks 4  3  Basal Antibiotic' Vitamins & Minerals Vitamins, Minerals & Antibiotic  1.91 1.85 1.86 .1.78  2.29 2.24 2.29 2.28  ~- i Feed e f f i c i e n c y 5 f o r test period 2.38 2.22 2.91 2.64  2.15 2.06 2.27 2.16  5  Feed e f f i c i e n c y f o r test period  2.78 2.39 3.51 2.62  2.53 2.12 2.54 2.33  Soybean R a t i o n Ration No. 5 6 7 8  Ration  3  Basal Antibiotic Vitamins & M i n e r a l s Vitamins, Minerals & Antibiotic  2.02 1.84 2.21 1.97  Fishmeal Ration No;  9  10  11 12  Ration Basal Antibiotic Vitamins & Minerals Vitamins, Minerals & Antibiotic  3 2.18 1.81 2.08 1.81  Weeks 4 2.91 2.29 2.34 2.59  Ration Weeks 4  2.55 2.19 2.06 2.13  5  Feed e f f i c i e n c y f o r t e s t period  2.55 2.30 2.62 2.33  2.38 2.05 2.23 2.05  T a b l e IV  Analysis of Variance  Source of Variation  Sum of Squares  Total 988,922 Rations 44,270 Antibiotics 227,897 Vitamins & Minerals 5,418 Rations x A n t i b i o t i c s 23,l6l Rations x Vitamins & Minerals 43,547 A n t i b i o t i c s x V i t a m i n s & M i n e r a l s 2,936 Rations x A n t i b i o t i c s x 79,762 Vitamins & Minerals Error 561,931  *  S i g n i f i c a n t a t p = 0.05 when t r i p l e  at f.  Variance  224 2 1 1 2 2 1 2  22,135 227,897* 5,418 11,580 21,773 2,936 39,881  ',. 231  .  2,639  interaction  ( r a t i o n s x a n t i b i o t i c s x v i t a m i n s and m i n e r a l s ) i s used as the t e s t i n g  error.  - 58 -  .  F i g u r e 1.  Basal Antibiotics V i t a m i n s and M i n e r a l s V i t a m i n s , M i n e r a l s and  Antibiotics  ".-Growth, r e s p o n s e of c h i c k s r e c e i v i n g supplements.  different  -  59  -  SOYBEAN RATION  600  r  Age i n weeks  Basal . . .. . ...... A n t i b i o t i c s . Vitamins and M i n e r a l s v i t a m i n s , M i n e r a l s and A n t i b i o t i c s  F i g u r e 1. - "^Growth tr.esponsfe; of c h i c k s r e c e i v i n g d i f f e r e n t supplements.  FISHMEAL  RATION  600  CO  450  S cd  c «H  •P  300  •H <D  150  2  3 Age i n weeks  Basal Antibiotics Vitamins and M i n e r a l s V i t a m i n s , M i n e r a l s and A n t i b i o t i c s F i g u r e 1.  Growth response of c h i c k s d i f f e r e n t supplements.  receiving  - 61 -  Experiment  11  Introduction The second study was  conducted t o determine  whether o r not a n t i b i o t i c s would lower the p r o t e i n ment o f the c h i c k .  Rations containing  require-  different protein  l e v e l s were f o r m u l a t e d w i t h the same b a s i c i n g r e d i e n t s i n each r a t i o n .  The p r o t e i n l e v e l s were lowered by i n c r e a s i n g  the ground wheat and ground y e l l o w c o r n and  decreasing  the soybean o i l meal and f i s h m e a l used i n the r a t i o n s . Two  d i f f e r e n t l e v e l s of a n t i b i o t i c s were t e s t e d w i t h these  basal r a t i o n s .  A l l r a t i o n s were f e d t o both male and female  c h i c k s t o determine sex d i f f e r e n c e i n growth response t o antibiotics. TEST NO.  2  ' Purpose The purpose o f t h i s t e s t was  t o determine the  e f f e c t o f a n t i b i o t i c s on the growth r a t e o f c h i c k s when f e d i n r a t i o n s w i t h a lower p r o t e i n l e v e l than t h a t  recommended  f o r optimum growth. Methods and Procedure Day o l d , female, White Leghorn c h i c k s were used i n this-experiment.  Twenty c h i c k s were d i s t r i b u t e d a t random  to each o f s i x t e e n brooder compartments.  The equipment  used  - 62 -  and  the method of h a n d l i n g  o u t l i n e d i n Test 1. The  the c h i c k s was the same as  Each r a t i o n was t e s t e d i n d u p l i c a t e .  b i r d s were weighed a t 7, 14,  29,  41 and 54 days and the  amount of f e e d consumed was recorded f o r the t e s t  period.  A r e c o r d o f the average weights i s presented i n Table and  the food  consumption i n Table 111.  days are presented i n F i g u r e  1.  11.  The weights a t 54  A s t a t i s t i c a l analysis  also  was made on the weights o f the c h i c k s a t 54 days and the r e s u l t s are presented i n Table IV. Four b a s a l r a t i o n s were used f o r t h i s t e s t and the p r o t e i n l e v e l f o r each r a t i o n adjusted protein levels.  16,  18 and 20%  A chemical a n a l y s i s of these r a t i o n s  showed the p r o t e i n l e v e l s t o be 16.0, respectively.  t o 14,  17.9,  19.5,  and 22.0  The i n g r e d i e n t s used, i n these r a t i o n s are  shown i n T a b l e 1.  E q u a l p a r t s of wheat and corn were used  as c e r e a l g r a i n s .  The p r o t e i n supplements were soybean  and  f i s h m e a l which were a l s o added i n equal p a r t s ;  decrease the p r o t e i n l e v e l the c e r e a l g r a i n s were and  the p r o t e i n supplements decreased.  To increased  As the p r o t e i n  was lowered, the amount o f bonemeal was i n c r e a s e d  level  due t o the  i t was f a c t t h a t the f i s h m e a l was reduced and hence / n e c e s s a r y t o adjust  the calcium-phosphorus r a t i o .  Vitamins were added  to each r a t i o n i n the same amounts r e g a r d l e s s level.  o f the p r o t e i n -  - 63 -  The e f f e c t o f adding an a n t i b i o t i c was t e s t e d w i t h 0.5% o f an a n t i b i o t i c p r e p a r a t i o n c o n t a i n i n g 1.8 of aureomycin  gms. per l b .  hydrochloride.  Results The weights a t 54 days showed t h a t those b i r d s on the 16% l e v e l o r p r o t e i n averaged 536 grams without aureomycin and 586 grams w i t h aureomycin o r an i n c r e a s e o f 5.6% 17.9%  due t o the e f f e c t of aureomycin.  The c h i c k s on the  p r o t e i n l e v e l averaged 662 grams without aureomycin  and 712 grams w i t h aureomycin o r an i n c r e a s e o f 7.5% due t o the e f f e c t o f aureomycin.  The c h i c k s on the 19.5%  level  averaged 748 grams without aureomycin and 728 grams w i t h aureomycin showing a decrease i n weight when aureomycin was added.  The c h i c k s on the 22% l e v e l averaged 770  grams  without aureomycin and 784 grams w i t h aureomycin o r an i n c r e a s e o f 1.9%  due t o the e f f e c t o f aureomycin.  I t i s e v i d e n t from the growth data t h a t the a d d i t i o n o f aureomycin t o the b a s a l r a t i o n s c o n t a i n i n g the different protein levels the c h i c k f o r p r o t e i n .  d i d not lower the requirement of Although a g r e a t e r response was  obtained by adding aureomycin t o the r a t i o n s a t the 16 and 17.9%  protein levels  than a t the 1 9 . 5 and 22% l e v e l s ,  still  i n no i n s t a n c e d i d the a n t i b i o t i c b r i n g the weights up t o the weights o f the c h i c k s a t the 22% l e v e l .  The s t a t i s t i c a l  - 64 a n a l y s i s on the average weights a t 54 days showed t h a t  there  were no s i g n i f i c a n t d i f f e r e n c e s due t o the a d d i t i o n of the a n t i b i o t i c t o the d i f f e r e n t r a t i o n s , the o n l y r e a l d i f f e r e n c e s being due t o the v a r i e d p r o t e i n l e v e l s . The  f e e d e f f i c i e n c y r e s u l t s showed a lowered f e e d  u t i l i z a t i o n a t the lower p r o t e i n l e v e l . aureomycin t o the 16 and 17.9%  The a d d i t i o n o f  p r o t e i n r a t i o n s d i d not  improve the f e e d e f f i c i e n c y over the b a s a l r a t i o n s . f e d a t the 17.9  and 19.5%  Rations  l e v e l s were equal i n v a l u e w i t h  and without aureomycin i n so f a r as feed u t i l i z a t i o n was concerned. Conclusions 1.  When the p r o t e i n l e v e l of c h i c k r a t i o n s was lowered, i t was found t h a t the a d d i t i o n of an a n t i b i o t i c increased  growth response a t lower l e v e l s  but n o t s u f f i c i e n t l y t o b r i n g the average weights up t o those a t the 22% p r o t e i n l e v e l . t h a t adding an a n t i b i o t i c t o r a t i o n s  This  showed  containing  lower l e v e l s o f p r o t e i n does not lower the requirement of the c h i c k f o r p r o t e i n . 2.  Feed e f f i c i e n c y was lessened levels.  w i t h the lower p r o t e i n  The a d d i t i o n of aureomycin d i d not improve  the e f f i c i e n c y a t any o f the l e v e l s .  - 65  -  1  Table  Composition of B a s a l R a t i o n s  16% Ingredient  j  17.9%  Basal  Basal  pounds Ground wheat Ground y e l l o w corn Soybean o i l meal Fishmeal Dehydrated c e r e a l grass *BY Iodized s a l t Feeding o i l (2250A: 3 0 0 D ) . Ground limestone Bonemeal Manganese sulphate C h o l i n e c h l o r i d e (25%) Nicotinic acid Calcium pantothenate  41.1 41.1 3.7 3.7 5.0 1.0 0.5 .0.25 1.0 2.0 0.025 0.4 0.8  0.5  19.5%  gm. gm.  Basal  per  39.0 39.0 6.0 6.0 5.0 1.0 0.5 0.25 1.0 1.75  0.025  0.4 0.8 0.5  gm. gm.  Rations #1 - #4 i n c l u s i v e - b a s a l s . " # l a - #4a i n c l u s i v e - b a s a l s plus 0.5% aurofac.**  * **  BY - commercial product supplement.  used mainly  100  22% Basal  pounds  37.2 37.2 8.0 8.0 5.0 1.0 0.5 0.25 1.0 1.25 0.025 0.4 0*8 gm. 0 . 5 gm.  -  35.4 35.4 10.0 10.0 5.0 1.0 0.5 0.25 1.0 1.0 0.025 0.4 0 . 8 gm. 0.5 gm.  aureomycin  for riboflavin  Aureomycin aurofac - commercial p r e p a r a t i o n c o n t a i n i n g aureomycin h y d r o c h l o r i d e as an a n t i b i o t i c .  - 66 -  Table  11  Average Weights  it i o n No.  1 la  2 2a 3 3a 4 4a  Protein level-%  16.0 16.0 17.9 17.9 19.5 19.5 22.0 22.0  14  7 69 71 80  83 84 84 83 86  gm.  105 gm. 109 130 138 138 140 • 138 144  Table  Days  29  41  224 gm. 250 299 311 326 321 240 338  376 gm. 410 478 499 524 510 540 549  54  532 586 662 712 748 728 770 784  111  Feed E f f i c i e n c y  R a t i o n No.  1 la  2 2a 3  Feed e f f i c i e n c y f o r test period  3.0 3.2 2.5 2.8 2.5  - 67 -  T a b l e IV  Analysis of Variance  Source of variation  Sum of Squares  Total 3,480,844 Blocks 30,275 Antibiotics 34,175 Protein 2,081,198 Blocks x A n t i b i o t i c s 413 Blocks x Protein 3,911 Antibiotic x Protein 74,854 Blocks x A n t i b i o t i c x P r o t e i n 110,042 Error 1,145,976  *  S i g n i f i c a n t a t p = 0 . 0 1 when t r i p l e  d.f.  Variance  313 1 1 3 1 3 3 3 298  30,275 34,175 693,732* 413 1,303 24,931 36,680 3,846  interaction  ( b l o c k s x a n t i b i o t i c x p r o t e i n ) i s used as the t e s t i n g  error.  - 68 -  FINAL AVERAGE WEIGHTS 54 DAYS  1..BASAL-16% PROT. ' l a . #1 4 ANTIB. "2. BASAL-17.9% PROT. 2a. #2 4 -ANTIB. 3. BASAL-19.5% PROT. -3a. #3 + ANTIB. 4 . BASAL-22.0% PROT. -2,a. #4 + ANTIB. 0  F i g u r e 1.  500  600 GRAMS  700  Average weights of c h i c k s f e d 16, 1 7 . 9 , 1 9 . 5 and 22.0 per cent p r o t e i n r a t i o n s supplemented with a n t i b i o t i c s .  800  - 69 -  TEST NO,  3  Purpose The purpose o f t h i s experiment,was results obtained  i n T e s t No.  l e v e l o f the r a t i o n .  t o c o n f i r m the  2 by again l o w e r i n g the p r o t e i n  Aureomycin was  again added t o the b a s a l  r a t i o n s t o determine i f i t had any e f f e c t on lowering the p r o t e i n needs of the c h i c k . Methods and Procedure Day o l d , female, New Hampshire t h i s experiment.  c h i c k s were used i n  D i s t r i b u t i o n and h a n d l i n g o f the b i r d s  the same as i n the previous t e s t s . The c h i c k s were weighed  was  A l l t e s t s were i n d u p l i c a t e . each week t o the end of the  f o u r t h week; a r e c o r d o f t h e i r average' weights i s shown i n Table 1.  Feed consumption was kept f o r the t r i a l p e r i o d and  the feed e f f i c i e n c y i s  recorded  i n Table  11.  The i n g r e d i e n t s i n the b a s a l r a t i o n s used i n t h i s experiment were the same as i n Test No.  2.  Two  were made up w i t h p r o t e i n l e v e l s at 22 and 16%. b a s a l was added 0.5% 0.9 gms.  basal rations To each  o f an a n t i b i o t i c p r e p a r a t i o n c o n t a i n i n g  o f aureomycin  hydrochloride.  Results The r e s u l t s i n growth from t h i s experiment showed t h a t the c h i c k s f e d a t the 22% l e v e l without aureomycin  - 70 -  averaged 351 grams and with aureomycin averaged 352 grams. In other words, no increase was to he noted from the use of an a n t i b i o t i c i n t h i s ration.  The chicks fed at the 16%  l e v e l without aureomycin averaged 280 grams and with aureomycin averaged 294 grams or 5% increase.  These r e s u l t s ,  therefore* confirm those obtained i n Test No. 2 i n that although a greater response was obtained at the lower protein l e v e l , with the addition of a n t i b i o t i c s , there was not a lowering of the requirement of the chick f o r protein. The addition of a n t i b i o t i c s had l i t t l e or no effect on feed e f f i c i e n c y i n t h i s experiment.  Efficiency  was lower at the lower protein l e v e l . Conclusions This test was conducted to confirm the results of Test No. 2.  The data again showed that the addition of  a n t i b i o t i c s did. not lower the protein requirements of the chick.  - 71 -  Table 1  Average  Weights Weeks  Ration No. 1 la 2 2a  Protein level-%  1 82 83 73 74  22.0 22.0 16.0 16.0  Table  Feed  R a t i o n No. 1 la 2 2a  2 155 155 125 130  3  4  237 242 192 201  351 352 280 294  11  Effioiency Feed e f f i c i e n c y f o r test period 2.17 2.12 2.52 2.57  R a t i o n 1 and 2 - b a s a l s Rations l a and 2a - b a s a l s plus 0.5% aureomycin  aurofac.  - 72 -  TEST NO. 4  Purpose The purpose o f t h i s t e s t was a g a i n t o study the e f f e c t o f a n t i b i o t i c s on a lowered a d d i t i o n t o determine  p r o t e i n i n t a k e and i n  the e f f e c t o f a n t i b i o t i c s as t o sex.  The response when a h i g h e r l e v e l o f a n t i b i o t i c was f e d i n the r a t i o n was a l s o s t u d i e d . Methods and Procedure Day o l d , male and female, White Leghorn c h i c k s were used i n t h i s experiment  and the method o f d i s t r i b u t i o n  and h a n d l i n g was the same as i n the previous The  tests.  composition o f the r a t i o n s were the same as i n  the previous t e s t s  and were f e d a t 16 and 22% p r o t e i n  levels.  An a n t i b i o t i c was added t o the b a s a l r a t i o n s a t 0.5% and 1.0% l e v e l s t o determine  the e f f e c t o f a h i g h e r l e v e l o f f e e d i n g .  A g a i n an aureomycin p r e p a r a t i o n was used as i n the p r e v i o u s tests. The  c h i c k s were weighed a t the end o f each week up  to the end o f f o u r weeks and the r e s u l t s T a b l e 1 and the f o u r t h week r e s u l t s  a r e presented i n  i n F i g u r e 1.  Feed  e f f i c i e n c y data was recorded and i s presented i n Table 11.  - 73 -  Results The r e s u l t s  o f the c h i c k s weights  a t f o u r weeks o f  age show t h a t the female c h i c k s f e d the 22% b a s a l  ration  averaged 366 grams and t h e males averaged 405 grams without antibiotic.  When the a n t i b i o t i c was added a t the 0.5% l e v e l ,  the females averaged 399 grams o r 9.01% i n c r e a s e over those without a n t i b i o t i c .  When added a t the same l e v e l , the males  averaged 436 grams o r a 7.6% i n c r e a s e over those without antibiotic.  a t t h e 1.0%  The female c h i c k s f e d a n t i b i o t i c  l e v e l averaged 403 grams o r 10.1% i n c r e a s e over those without a n t i b i o t i c .  The male c h i c k s f e d a n t i b i o t i c  chicks  a t the  same l e v e l averaged 436 grams o r 13.5% i n c r e a s e over those c h i c k s without a n t i b i o t i c .  These r e s u l t s  suggest t h a t t h e r e  i s no d i f f e r e n c e i n growth by sexes when a n t i b i o t i c s are added.  The h i g h e r l e v e l o f aureomycin  growth i n e i t h e r  sex.  protein basal ration  The female c h i c k s f e d on t h e 16%  averaged 282 grams without  and t h e males averaged 297 grams. antibiotic  had no added e f f e c t on  antibiotic  The a d d i t i o n o f 0.5%  r e s u l t e d i n average weights o f 309 grams i n the  females o r a 9.5% i n c r e a s e over those c h i c k s without antibiotic.  The a d d i t i o n o f 0.5% a n t i b i o t i c  average weights  resulted i n  o f 337 grams i n the males o r a 13.5%  over those c h i c k s without a n t i b i o t i c .  When 1.0%  increase  antibiotic  was added the female c h i c k s averaged 305 grams o r a 8.1% i n c r e a s e over those c h i c k s without a n t i b i o t i c .  When 1.0%  - 74 -  a n t i b i o t i c was added the male c h i c k s averaged 336 grams o r a 13.1% i n c r e a s e over those c h i c k s without a n t i b i o t i c .  The  r e s u l t s here again are the same as those o b t a i n e d when 22% p r o t e i n was f e d . The a d d i t i o n o f a n t i b i o t i c s t o the b a s a l r a t i o n s improved  the f e e d e f f i c i e n c y t o some extent except when 0.5%  a n t i b i o t i c was f e d t o the 16% b a s a l . some e r r o r i n t h i s  There appears t o be  result.-  Conclusions 1.  Adding an a n t i b i o t i c t o a r a t i o n i n which the p r o t e i n l e v e l was lowered t o 16% r e s u l t e d i n no e f f e c t on l o w e r i n g the p r o t e i n needs o f the c h i c k when compared t o a r a t i o n c o n t a i n i n g 22% p r o t e i n .  2.  There was l i t t l e  d i f f e r e n c e i n response t o growth  between sexes when an a n t i b i o t i c was added.  The  males appeared t o grow f a s t e r than the females a t the 16% p r o t e i n l e v e l when compared w i t h the growth response a t the 22% p r o t e i n l e v e l when a n t i b i o t i c was added t o both. 3.  I n c r e a s i n g t h e a n t i b i o t i c l e v e l from 0.5 t o 1.0% d i d not i n c r e a s e the r a t e o f growth. again as i n p r e v i o u s t e s t s was  The a n t i b i o t i c  aureomycin.  - 75 -  Table  1  Average Weights Ration P r o t e i n No. 1 2 3 4 5 6  Level-% 22 22 22 16 16 16  Weeks 1 2 3 4 Female Male Female Male Female Male Female Male 75 77 78 64 70 67  79 84 84 66 71 73  201 206 216 146 161 153  Table  Feed  R a t i o n No. 1 2 3 4 5 6  204 218 226 142 163 167  265 283 291 201 228 220  274 305 304 202 238 242  366 399 403 282 309 305  11  Efficienoy  Feed e f f i c i e n c y f o r test period 2.35 2.63 2.22 2.97 2.81 2.75  Rations 1 and 4 - b a s a l s . Rations 2 and 5 - b a s a l s plus 0 . 5 % a u r o f a c . Rations 3 and 6 - b a s a l s plus 1.0% a u r o f a c .  - 405 436 436 297 337 336  - 76 -  FINAL AVERAGE WEIGHTS' AT 1+ WEEKS  1. BASAL-22% PROTEIN  •'F  1. BASAL-22% PROTEIN  M  2. #1 PLUS 0.5% ANTIBIOTIC  F  2. #1 PLUS 0.5% ANTIBIOTIC  M  3. #1 PLUS 1.0% ANTIBIOTIC  F  ///////  3. #1 PLUS 1.0% ANTIBIOTIC  •M  ////////  4. BASAL-16% PROTEIN  F  4. BASAL-16% PROTEIN •  M  5. #4 PLUS 0.5% ANTIBIOTIC  F  5. #4 PLUS 0.5% ANTIBIOTIC  M  6. #4 PLUS 1.0% ANTIBIOTIC  F  6. #4 PLUS 1.0% ANTIBIOTIC  M  >///////rn /////// m> ///////////\  0  250  300  7~7~7~A  350  400 450  GRAMS  Figure  1.  Average weights o f male and female c h i c k s f e d 22% and 16% p r o t e i n r a t i o n s supplemented w i t h two d i f f e r e n t l e v e l s o f a n t i b i o t i c s .  - 7-7-  Experiment  111  Introduction The t h i r d study was designed t o determine whether or not a n t i b i o t i c s would reduce the requirement o f the c h i c k f o r s p e c i f i c amino a c i d s .  The e f f e c t was determined o f  a n t i b i o t i c s on c h i c k growth i n a b a s a l r a t i o n wheat as the main i n g r e d i e n t . lysine.  composed of  T h i s r a t i o n was d e f i c i e n t i n  A g a i n the e f f e c t o f a n t i b i o t i c s on c h i c k growth i n  a tryptophan d e f i c i e n t  b a s a l r a t i o n was determined by u s i n g  corn as the main i n g r e d i e n t i n the r a t i o n .  Crystalline  l y s i n e and tryptophan were added t o the r e s p e c t i v e  basal  r a t i o n s t o determine t h e i r e f f e c t on growth response w i t h and without the a d d i t i o n o f a n t i b i o t i c s . ,, TEST NO. 5 Purpose 1.  To study the e f f e c t o f a n t i b i o t i c s on growth i n  c h i c k s when added t o a wheat r a t i o n which i s d e f i c i e n t i n l y s i n e and a l s o the e f f e c t when added t o a corn r a t i o n which is deficient 2.  i n tryptophan.  To compare  t h i s growth response t o the b a s a l r a t i o n s  t o which have been added c r y s t a l l i n e amino a c i d s t o make up the  deficiencies.  - 7'8 -  3.  To compare two a n t i b i o t i c s i n so f a r as t h e i r  growth promoting  s t r e n g t h i s concerned  when added t o the b a s a l  rations. ' Methods and Procedure Day  o l d , male, New Hampshire c h i c k s were used i n  these experiments  which were d i s t r i b u t e d and handled as i n  the previous t e s t s .  Twenty c h i c k s were d i s t r i b u t e d t o each  compartment. The  c h i c k s were weighed a t weekly i n t e r v a l s and  the experiment  was c a r r i e d f o r t h r e e weeks.  growth response  Results of the  are p r e s e n t e d i n Table 1 1 and F i g u r e 1 .  s t a t i s t i c a l analysis  was made on the average weights  A  a t three  weeks and the r e s u l t s are shown i n Table 1 1 1 . The wheat r a t i o n was comprised  o f 88.83 pounds, o f  ground wheat t o which was added 5 pounds o f c e r e a l grass t o i n c r e a s e the f i b r e , a n d ^ p r o t e i n t o p r o v i d e some o f the e s s e n t i a l v i t a m i n s and m i n e r a l s .  The m i n e r a l s were balanced  by adding 1 pound o f limestone and 2 pounds o f bonemeal. The v i t a m i n s were balanced by adding c h o l i n e , v i t a m i n s A and D, r i b o f l a v i n (BY), n i c o t i n i c a c i d , pantothenic a c i d and vitamin B12.  T h i s r a t i o n , however, was d e f i c i e n t i n the  animo a c i d l y s i n e .  The corn r a t i o n was made up o f 80.83 l b .  of ground corn along w i t h the same i n g r e d i e n t s t h a t were used i n the wheat r a t i o n except t h a t n i c o t i n i c a c i d was omitted.  - 79 -  The reason f o r t h i s was  due to the f a c t t h a t without  n i c o t i n i c a c i d , tryptophan S i n c e tryptophan was n i c o t i n i c a c i d was  cannot be u t i l i z e d i n the body.  the d e f i c i e n t amino a c i d being s t u d i e d ,  omitted.  Ten  pounds of g e l a t i n  added t o t h i s r a t i o n t o r a i s e the p r o t e i n the o t h e r amino a c i d s . r a t i o n was chemical  14.79% and  The  protein  by  analysis.  1 - l y s i n e HC1  and w i t h 1.5  gm.  corn r a t i o n (#5) ( r a t i o n #7) gm.  balance  18.37% f o r the corn as determined  ( r a t i o n #2)  again supplemented w i t h 1.0%  1.5  l e v e l and  l e v e l f o r the wheat  The wheat b a s a l r a t i o n (#1) 0.5%  was  per 100  was  supplemented w i t h  lb.  R a t i o n #1  aureomycin aurofac ( R a t i o n  of p e n i c i l l i n ( R a t i o n #4) was  per 100  supplemented w i t h 0.2%  and again w i t h 1.0%  per 100  lb.  lb.  To t h i s r a t i o n was  d l - t r y p t o p h a n ( r a t i o n #10) aurofac ( r a t i o n #11)  and again 1.0%  and 1.5  gm.  The  and  In r a t i o n  added t o the corn r a t i o n to study  e f f e c t on c h i c k growth.  #3)  dl-tryptophan  aureomycin a u r o f a c  of p e n i c i l l i n ( r a t i o n #8)  n i c o t i n i c a c i d was  was  added  #9  the  0.2%  o f aureomycin  of p e n i c i l l i n ( r a t i o n  #12)  per 100 l b . Results The  average weights a t f o u r weeks f o r the wheat  r a t i o n showed the c h i c k s f e d the b a s a l r a t i o n (#1) 91 grams.  Those f e d d l - l y s i n e ( r a t i o n #2)  weighed  weighed 131  gm.  -  80 -  while those f e d aureomycin weighed 109 weighed 106  gm.  gm.  and  penicillin  These r e s u l t s show t h a t although the  a n t i b i o t i c s d i d improve the b a s a l r a t i o n (#1),  the improvement  was not s i g n i f i c a n t s i n c e they f a i l e d t o reach the weight o f the  amino a c i d supplemented r a t i o n (#2).  The average weights  f o r the corn r a t i o n showed t h a t the c h i c k s f e d the b a s a l r a t i o n (#5)  weighed  57 grams w h i l e the tryptophan supplemented  r a t i o n (#6)  weighed 83 grams.  Again, however, the c h i c k s  supplemented w i t h aureomycin (#7)  weighed o n l y 59 grams and  those f e d p e n i c i l l i n (#8) weighed 60 grams. interpretation addition  as i n the wheat r a t i o n must be concluded.  The  o f . n i c o t i n i c a c i d i n c r e a s e d the average weights t o  65 gm. when f e d w i t h the c o r n r a t i o n (#9). of tryptophan (#10) of 116  The same  grams.  resulted  addition  a g a i n improved growth w i t h average weights  Aureomycin  ( r a t i o n #11)  fed to ration  #9  i n average weights o f 73 grams w h i l e p e n i c i l l i n  ( r a t i o n #12) 75 grams.  The  f e d t o r a t i o n #9 r e s u l t e d  i n average weights of  Again, however, although the a d d i t i o n  o f the  a n t i b i o t i c s improved the average weights they were not e q u a l to the tryptophan supplemented r a t i o n .  The a d d i t i o n  of  n i c o t i n i c a c i d , however, d i d improve average weights i n d i c a t i n g i t s s y n e r g i s t i c e f f e c t w i t h tryptophan.  The  addition  of p e n i c i l l i n o r aureomycin t o the b a s a l  rations  resulted  i n equal responses t o growth i n d i c a t i n g t h a t t h e i r  v a l u e as a n t i b i o t i c s under these c o n d i t i o n s were the same.  A s t a t i s t i c a l a n a l y s i s on the weights at t h r e e weeks of age  showed the d i f f e r e n c e s t h a t were s i g n i f i c a n t  f o r the d i f f e r e n t treatments. was  No  significant difference  o b t a i n e d when a n t i b i o t i c s were added t o the corn b a s a l  ration  (#5). M o r t a l i t y was  r e s u l t e d i n 16 deaths.  h i g h i n the corn b a s a l r a t i o n When tryptophan was  (#5)  added (#6)  there  were no deaths w h i l e o n l y 4 deaths o c c u r r e d i n the aureomycin f e d r a t i o n . ( # 7 ) and the same number i n the p e n i c i l l i n f e d ration (#8).  T h i s i n d i c a t e s t h a t the a n t i b i o t i c s have a  value i n d e c r e a s i n g m o r t a l i t y .  Only one  the c h i c k s when n i c o t i n i c a c i d was and supplemented r a t i o n s (#9,  10,  death o c c u r r e d among  added t o the corn b a s a l 11,  12).  0onelusions 1.  The  a d d i t i o n o f a n t i b i o t i c s to wheat and  r a t i o n s d e f i c i e n t i n l y s i n e and  corn  tryptophan  r e s p e c t i v e l y had no e f f e c t i n l o w e r i n g the need of these amino a c i d s f o r the c h i c k . 2.  There" was  no  d i f f e r e n c e between p e n i c i l l i n  and  aureomycin i n t h e i r growth promoting e f f e c t under the c o n d i t i o n s of t h i s 3.  experiment.  A n t i b i o t i c s appeared t o have an e f f e c t on r e d u c t i o n of m o r t a l i t y i n the corn, tryptophan diet.  deficient  Table  1  Composition of B a s a l  Wheat B a s a l #1 lb./lOO l b .  Ingredient Ground wheat Ground corn Gelatin Dehydrated c e r e a l grass Ground limestone Bonerneal Iodized s a l t Manganese sulphate C h o l i n e c h l o r i d e (25%) Feeding o i l (2250A: 300D) BY Nicotinic acid Calcium pantothenate V i t a m i n B]_2  Ration  #1 #2  #3  . n  #5 #6  n  it  n  P  #8  #9  #10 #11 #12  Rations  basal. #1 plus #1 plus #1 plus basal. #5 Plus #5 plus #5 plus #5 plus #5 plus #5 plus #5 plus  Corn B a s a l #5 lb./lOO l b .  88.83 —  80.83 10.0  5.0  5.0  1.0 2.0  1.0 2.0  0.5  0.5 0.025 0.4 0.25  0.025  0.4  0.25 2.0 0.8  gm. 0.5 gm. 0.001 gm.  1 - l y s i n e HC1. aureomycin. penicillin. dl-tryptophan. aureomycin. penicillin, n i c o t i n i c acid. n i c o t i n i c a c i d & dl-tryptophan. n i c o t i n i c a c i d & aureomycin. nicotinic acid & p e n i c i l l i n .  2.0 .-  0.5  0.001  gm. gm.  Table  11  Average weights a t 3 weeks  R a t i o n No.  1 2 3 4 5 6 7 8 9 10 11 12  Ration  Average weight  Wheat Wheat plus l y s i n e Wheat plus aureomycin Wheat plus p e n i c i l l i n Corn Corn p l u s tryptophan Corn p l u s aureomycin Corn plus p e n i c i l l i n Corn p l u s n i c o t i n i c a c i d Corn plus n i c o t i n i c a c i d &. tryptophan Corn p l u s n i c o t i n i c a c i d & aureomycin Corn p l u s n i c o t i n i c a c i d & p e n i c i l l i n  Table  91 131 109 106 57 83 59 60 65. 116 73. 75  111  A n a l y s i s o f Variance Source of V a r i a t i o n  Total  Treatment  Error  Sum o f - Squares  d.f.  157,544  211  41,521  200  116,023  11  S i g n i f i c a n t a t the Ifo l e v e l .  variance  10,548* 208  Minimum Significant Difference  13  10  - 84 -  FINAL AVERAGE WEIGHTS AT 3 WEEKS  v///////k  1. WHEAT 2.  WHEAT + LYSINE  3.  WHEAT + AUREOMYCIN  4.  WHEAT + PENICILLIN  5.  CORN  6.  CORN + TRYPTOPHAN  7.  CORN + AUREOMYCIN  8.  CORN + PENICILLIN  9.  CORN + NICOTINIC ACID  10.  #9  11.  #9 +  AUREOMYCIN  12.  #9 +  PENICILLIN  7ZZZZZZZZZZZ /////////J  /////////  4 TRYPTOPHAN  0  .  50  75  100  125  GRAMS  F i g u r e 1.  Average weights o f c h i c k s f e d wheat and c o r n r a t i o n s supplemented w i t h amino a c i d s , a n t i b i o t i c s and a v i t a m i n .  150  - 35 -  TEST NO.  6  Purpose The purpose o f t h i s experiment was  t o determine the  e f f e c t on chicle growth o f adding a supplement o f v i t a m i n s t o a wheat r a t i o n d e f i c i e n t i n l y s i n e .  A l s o t o determine the  e f f e c t of these v i t a m i n s w i t h l y s i n e and/or added t o the b a s a l  penicillin  ration.  Methods and Procedure Day o l d , male, New Hampshire  c h i c k s were used i n  t h i s experiment and were d i s t r i b u t e d and handled as i n previous experiments. The experiment was  c a r r i e d out f o r t h r e e weeks and  the. c h i c k s weighed at the end o f each week. average weights are p r e s e n t e d i n T a b l e 1.  A  A r e c o r d o f the statistical  a n a l y s i s was made on the weights a t 3 weeks and the a n a l y s i s i s shown i n Table 11. the  Feed consumption was  recorded f o r  t e s t p e r i o d and the f e e d e f f i c i e n c y r e c o r d i s a l s o  presented i n Table 1. The b a s a l r a t i o n used i n t h i s experiment  was  composed o f the same i n g r e d i e n t s as i n the p r e v i o u s t e s t . R a t i o n s were supplemented w i t h 0.5% d l - l y s i n e HC1 A v i t a m i n supplement p y r i d o x i n e , 5.0 gm.  composed of 0.05  gm.  per 100 l b .  thiamin, 0.10  para-amino benzoic a c i d ,  0.025 gm.  gm. folic  - 86 -  a c i d , 0.025 gm. 2-4 naphthoquinone, b i o t i n per 100 l b . was used.  5.0 gm. i n o s i t o l ,  0.01 gm.  P e n i c i l l i n was f e d a t the r a t e  of 1.5 mg. p e r 100 l b . Results The average weights of the c h i c k s a t 3 weeks o f age showed t h a t the c h i c k s on the basal- wheat r a t i o n (#1)  weighed  92 grams w h i l e those c h i c k s on the r a t i o n t o which l y s i n e was added (§2) averaged 115  grams.  When the v i t a m i n s were added  alone t o the b a s a l (#3). the c h i c k s weighed  93 grams o r no  i n c r e a s e over the weights o f the c h i c k s on the b a s a l r a t i o n . When added w i t h l y s i n e , an average o f 136 grams.  (#4), however, the weights i n d i c a t e d (#5)  The a d d i t i o n o f p e n i c i l l i n  r e s u l t e d i n no improved growth w i t h average weights o f 91 grams. P e n i c i l l i n and l y s i n e combined  (#6) r e s u l t e d i n a s u b s t a n t i a l  i n c r e a s e w i t h an average weight, o f 154 grams. combined w i t h the v i t a m i n supplement weighing 102 grams on an average.  (#7)  Penicillin  r e s u l t e d i n chicks  When l y s i n e , v i t a m i n s and  p e n i c i l l i n were added (§8) the average weights were 177 grams. The r e s u l t s from t h i s t e s t i n d i c a t e t h a t n e i t h e r the v i t a m i n supplement n o r p e n i c i l l i n when added i n combination. or alone r e s u l t e d / r e s u l t e ^ ) i n any growth i n c r e a s e .  When  l y s i n e was added t o e i t h e r the v i t a m i n supplemented r a t i o n (#3)  o r the p e n i c i l l i n r a t i o n (#5)  i n c r e a s e i n growth.  there was a s i g n i f i c a n t  The g r e a t e s t response was o b t a i n e d w i t h  the combination o f a l l three added t o the b a s a l (#8).  These .  r e s u l t s suggest, t h e r e f o r e , that i n order t o o b t a i n a response to e i t h e r a d d i t i o n a l v i t a m i n s  and/or p e n i c i l l i n ,  a c i d d e f i c i e n c y must be made up i n the r a t i o n .  the amino I t further  suggests t h a t n e i t h e r a v i t a m i n supplement nor an a n t i b i o t i c w i l l have any e f f e c t on lowering  the needs o f s p e c i f i c amino  a c i d s f o r the c h i c k . Feed e f f i c i e n c y data showed an improvement whenever l y s i n e was i n c l u d e d as a supplement t o the b a s a l r a t i o n . The  s t a t i s t i c a l a n a l y s i s on treatment o f r a t i o n s  showed t h a t only r e a l d i f f e r e n c e s were found when the l y s i n e d e f i c i e n t d i e t was supplemented w i t h c r y s t a l l i n e l y s i n e . Conclusions A vitamin  supplement and/or p e n i c i l l i n added t o  a wheat b a s a l d i e t had no e f f e c t on r e d u c i n g the amino a c i d , l y s i n e f o r the c h i c k .  the needs of  Table 1  Average Weights a t three weeks and Feed E f f i c i e n c y  Ration No. 1 2 3 4 5 6 7 8  Average Weights  • Ration Basal #1 plus #1 plus #1 plus #1 plus #1 plus #1 plus #1 p l u s  lysine vitamins l y s i n e and v i t a m i n s penicillin l y s i n e and p e n i c i l l i n v i t a m i n s and p e n i c i l l i n l y s i n e , vitamins & p e n i c i l l i n  Feed Efficiency  92 gm. 115 93 136 91 154 102 177  4.73 4.01 4.18 -2.55 4.88, 2.85 3.50 2.41  Table 11  Analysis of Variance  Source of Variation  Sum of Squares  d.f.  Variance  Total Treatment Error  192,077 148,982 43,095  158 7 151  21,283* 285  S i g n i f i c a n t a t the 1% l e v e l .  Minimum Significant Difference  10.57  13.95  -  89  -  TEST NO.  7  Purpose In t h i s experiment i t has  been assumed t h a t a r g i n i n e  i s a second l i m i t i n g amino a c i d i n a wheat r a t i o n . purpose was  The  t o t e s t the e f f e c t of a n t i b i o t i c s on growth of  c h i c k s when a r g i n i n e was  added t o a wheat b a s a l r a t i o n t o  which l y s i n e and a v i t a m i n supplement were i n c l u d e d . Methods and Day  f  Procedure  o l d , male, New  and handled as i n p r e v i o u s  Hampshire c h i c k s were d i s t r i b u t e d  experiments.  Twenty c h i c k s were  d i s t r i b u t e d to each of f o u r compartments. The  c h i c k s were kept f o r t h r e e weeks and weighed at  the end o f each week.  The  average weights over the  are presented  1.  Feed e f f i c i e n c y was  i n Table  the t r i a l p e r i o d as shown i n Table The  b a s a l r a t i o n ; used was  i n g r e d i e n t s as i n Tests HOI  was  11. composed of the same  5 and 6 except t h a t 0.5%  described  i n T e s t 5.  R a t i o n #2  supplemented w i t h p e n i c i l l i n at the r a t e of 15 #3  was  a l s o kept f o r  of  added along w i t h the. v i t a m i n supplement.  supplement was  supplemented w i t h 0.5%  1-lysine  This  was mg./lb.  a r g i n i n e and r a t i o n #4  supplemented with a combination of a r g i n i n e and the same l e v e l s as above.  period  Ration  was  p e n i c i l l i n at  D u r i n g the l a s t week of the  - 90 -  experiment 0.3% methionine was added t o r a t i o n s #3 and #4. Results The average weights a t three weeks o f age r e s u l t e d i n 131 grams f o r the c h i c k s f e d the b a s a l r a t i o n (#1). fed  the p e n i c i l l i n  Chicks  supplemented r a t i o n (#2) averaged 180 grams.  When a r g i n i n e was added t o the b a s a l r a t i o n (#3) the average weights were 115 grams.  T h i s showed t h a t a r g i n i n e d i d not  improve growth when used as a supplement.  R a t i o n #4,  t o which  p e n i c i l l i n and a r g i n i n e had been added, r e s u l t e d i n average weights o f 171 grams.  T h i s again showed t h a t a r g i n i n e had  no e f f e c t i n improving c h i c k growth when combined p e n i c i l l i n as a supplement. to a r g i n i n e i t was decided and #4  with  I n view o f the l a c k o f response t o add methionine t o r a t i o n s #3  during the l a s t week o f the experiment.  This  supplement, however, had no e f f e c t on c h i c k growth. Conclusions  '  A wheat b a s a l r a t i o n t o which had been added l y s i n e and a v i t a m i n supplement d i d not produce added growth a supplement o f a r g i n i n e and p e n i c i l l i n  with  over p e n i c i l l i n  alone.  T h i s showed t h a t a r g i n i n e was not a second l i m i t i n g f a c t o r when an a n t i b i o t i c was added.  Table  1  Average Weights  Ration No. 1 2 3 4  Ration  1  Basal #1 plus p e n i c i l l i n #1 p l u s a r g i n i n e #1 p l u s p e n i c i l l i n and a r g i n i n e  Table  Feed  R a t i o n No.  1 2 3 4  . Weeks 2  57 65 56 66  84 115 80 109  11  Efficiency  Feed e f f i c i e n c y f o r test period  4.04 3.15 4.56 3.25  3 131 180 115 171  - 92 Experiment  IV  Introduction The f o u r t h study was made t o determine what e f f e c t a n t i b i o t i c s might have i n lowering the carbohydrate requirement of the c h i c k . was  added t o a r a t i o n  I n the b a s a l r a t i o n ,  corn starch  composed o f ground y e l l o w c o r n and  ground wheat as the c e r e a l s and f i s h m e a l , soybean o i l meal and c e r e a l grass as the p r o t e i n carbohydrate l e v e l was  supplements.  lowered by r e p l a c i n g  with an equal amount of ground c e l l u l o s e . tests,  The  available  the corn s t a r c h As i n the  protein  two l e v e l s o f a n t i b i o t i c s were compared by c h i c k  growth response. c h i c k s was  also  The e f f e c t of a n t i b i o t i c s on male and female determined. TEST NO.  8  Purpose To study the e f f e c t of a n t i b i o t i c s on c h i c k s f e d a r a t i o n i n which the a v a i l a b l e  carbohydrates had been  lowered and t o determine whether or not a n t i b i o t i c s had  any  e f f e c t on l o w e r i n g the carbohydrate needs o f the c h i c k . Methods and Procedure Day o l d , male, New Hampshire c h i c k s were used i n t h i s experiment.  Twenty c h i c k s were d i s t r i b u t e d  at random  i n each of e i g h t compartments, the t e s t being run i n  duplicate.  - 93 -  H a n d l i n g o f the c h i c k s was the same as i n p r e v i o u s t e s t s . The experiment was c a r r i e d out over a f o u r week p e r i o d and the c h i c k s weighed a t the end o f each week. Average weights a t the end o f each Week a r e presented i n Table 1.  A r e c o r d of f e e d consumption was kept f o r the t r i a l  p e r i o d and the r e s u l t s a r e presented i n Table  11.  The same i n g r e d i e n t s were used i n the b a s a l r a t i o n as were used i n the p r o t e i n t e s t s .  The wheat and c o r n were  reduced and 15% c o r n s t a r c h was added t o make up the carbohydrate l e v e l ( r a t i o n #1).  The p r o t e i n l e v e l was  r e - a d j u s t e d by i n c r e a s i n g the soybean and f i s h m e a l .  In order  to lower the a v a i l a b l e carbohydrate l e v e l 15% ground c e l l u l o s e was s u b s t i t u t e d f o r the s t a r c h ( r a t i o n §2). preparation  {0.5%)  An aureomycin  was added t o both the s t a r c h r a t i o n and  the c e l l u l o s e r a t i o n . Results The b a s a l r a t i o n (#1)  t o which c o r n s t a r c h had been  added r e s u l t e d i n average weights o f 338 grams i n c h i c k s a t 4 weeks o f age.  The a d d i t i o n of aureomycin t o the b a s a l  (#3)  r e s u l t e d i n average weights of 350 grams o r an i n c r e a s e o f 3.5%.  The b a s a l r a t i o n (#2)  t o which ground c e l l u l o s e had  been added produced average weights of 33.6 grams. a d d i t i o n o f aureomycin t o t h i s b a s a l (#4)  The  resulted i n  average weights o f 346 grams o r an i n c r e a s e o f 2.9%.  These  r e s u l t s , t h e r e f o r e , i n d i c a t e t h a t a n t i b i o t i c s had no e f f e c t on  - 94 -  lowering the carbohydrate l e v e l requirements s i n c e the average weights ration  o f the c h i c k  o f the c e l l u l o s e and a n t i b i o t i c  (#4) f a i l e d t o reach the average weights  o f the s t a r c h  and a n t i b i o t i c r a t i o n (#3). The f e e d e f f i c i e n c y was lowered when 15% c e l l u l o s e was f e d i n the r a t i o n ration  (#1).  (#2) as compared t o the s t a r c h b a s a l  Some response.to a n t i b i o t i c s was noted however,  when t h i s r a t i o n  (#2) was supplemented w i t h aureomycin (#4).  Chicks on the h i g h e r carbohydrate r a t i o n and a n t i b i o t i c (#3) showed no improvement over the b a s a l s t a r c h r a t i o n (#1). Conclusions 1.  A n t i b i o t i c s had no e f f e c t on l o w e r i n g the carbohydrate needs o f the c h i c k when added t o " a r a t i o n low i n available  2.  carbohydrate.  Feed e f f i c i e n c y was lowered when the lower carbohydrate r a t i o n was f e d . antibiotics resulted ration.  The a d d i t i o n o f  i n some improvement i n t h i s  - 95 -  Table  1  Average Weights  Ration No. 1 2 3 4  Weeks 1  Ration Basal Basal Basal Basal  * **  - starch 84 gm. - alphacel* 85 #1 plus a n t i b i o t i o * * 8 6 #2 p l u s a n t i b i o t i c 87  2 152 148 157 156  3 gm.  234 gm. 226 244 239  Ground c e l l u l o s e . Aureomycin a u r o f a c ,  Table  11  Feed E f f i c i e n c y  R a t i o n No.  1 2 3 4  Feed e f f i c i e n c y f o r test period  1.99 2.47 1.98 2.25  4 338 336 350 346  gm.  - 96 -  •TEST NO.  9  Purpose To repeat the t e s t as o u t l i n e d i n T e s t No.  8 except  i n a d d i t i o n t o 0.5% of a n t i b i o t i c , a h i g h e r l e v e l (1.0%) a n t i b i o t i c was in this  fed.  of  Both male and female c h i c k s were used  experiment.  Methods and Procedure Day  o l d , male and female, White Leghorn c h i c k s were  used i n t h i s experiment.  The c h i c k s were d i v i d e d a t random  i n t o each o f 12 brooder compartments w i t h 15 c h i c k s i n each compartment.  The c h i c k s were handled the same as i n the  previous experiments. The b i r d s were, kept on experiment f o r f o u r weeks and weighed a t the end of each week.  A r e c o r d o f the average  weights i s presented i n T a b l e 1 and a graph showing the weights i s shown i n F i g u r e 1.  final  Feed consumption was r e c o r d e d  ifor the p e r i o d and the r e s u l t s are presented i n T a b l e 1 1 . The composition of the r a t i o n s was- .the same as i n the  p r e v i o u s te§t. a g a i n u s i n g 15% corn s t a r c h and 15% ground  c e l l u l o s e f o r the carbohydrate study. supplemented w i t h 0.5%  The b a s a l r a t i o n s were  aureomycin p r e p a r a t i o n and  1.0%  aureomycin p r e p a r a t i o n t o determine any d i f f e r e n c e i n growth response t o the h i g h e r l e v e l o f f e e d i n g .  - 97 -  Results The average weights o f the male and female c h i c k s f e d the s t a r c h b a s a l r a t i o n (#1) respectively.  were 380 grams and 394 grams  When 0.5% aureomycin p r e p a r a t i o n was added t o  t h i s b a s a l r a t i o n (#2) the females weighed 400 grams, o r an i n c r e a s e o f 5.2% and the males averaged 449 grams or an i n c r e a s e of 13.9%.  When 1.0% aureomycin p r e p a r a t i o n was f e d t o t h i s  b a s a l (#3) the female c h i c k s averaged 406 grams o r an i n c r e a s e of 6.8% over the b a s a l and 466 grams f o r the males o r an i n c r e a s e o f 17.5%.  The average weights o f the female and  male c h i c k s f e d the c e l l u l o s e b a s a l r a t i o n (#4) were 348 grams and 353 grams r e s p e c t i v e l y .  The a d d i t i o n o f 0.5% aureomycin  p r e p a r a t i o n t o t h i s b a s a l r a t i o n (#5)  r e s u l t e d i n average  weights o f 353 grams f o r the females o r 1.4% i n c r e a s e and 377 grams f o r the males o r 6.7% i n c r e a s e .  When 1.0% aureomycin  p r e p a r a t i o n was added t o t h i s b a s a l r a t i o n (#6) the females averaged 355 grams o r 2.01% i n c r e a s e and the males 368 grams or 4.2% i n c r e a s e .  The c h i c k s on the c e l l u l o s e b a s a l r a t i o n  supplemented w i t h a n t i b i o t i c d i d not weigh as much as the c h i c k s f e d the s t a r c h b a s a l and supplemented w i t h a n t i b i o t i c . T h i s again i n d i c a t e d t h a t a n t i b i o t i c s had no e f f e c t on l o w e r i n g the carbohydrate needs o f the c h i c k .  There appeared  to be a g r e a t e r response i n growth t o a n t i b i o t i c s from the males i n the s t a r c h f e d r a t i o n .  There was no d i f f e r e n c e  between the two l e v e l s o f a n t i b i o t i c f e d i n - s o - f a r as growth  98 -  was concerned. The f e e d e f f i c i e n c y r e s u l t s i n d i c a t e d  that  feed  e f f i c i e n c y was lowered when 15% ground c e l l u l o s e was added t o the  ration.  The e f f i o i e n o y was improved hy the a d d i t i o n o f  aureomycin i n each r a t i o n although no d i f f e r e n c e  was observed  between the two l e v e l s o f a n t i b i o t i c f e d . Conclusion 1.  Lowering the a v a i l a b l e chick ration,  carbohydrate l e v e l o f the  d i d not lower the carbohydrate needs  of the c h i c k when a n t i b i o t i c s were added. 2.  E f f i c i e n c y o f f e e d u t i l i z a t i o n was lowered when the carbohydrate l e v e l was lowered.  The a d d i t i o n o f  a n t i b i o t i c s improved the feed e f f i c i e n c y although no d i f f e r e n c e 3.  o c c u r r e d between the two l e v e l s f e d .  Male c h i c k s showed more growth response t o a n t i b i o t i c s than female c h i c k s when the 15% s t a r c h was f e d .  basal  ration  - 99 -  Table  1  Average Weights  Ration No.  Ration  Sex  1  Basal - starch  2  B a s a l #1  plus 0.5%  antibiotic  3  B a s a l #1  p l u s 1.0%  antibiotic  4  Basal - alphacel  5  B a s a l #4 p l u s 0.5%  antibiotic  6  B a s a l #4 plus 1.0%  antibiotic  F* M* F M F M F M F M F M  1 76 75 77 86 81  85 73 70 73 79 75 78  Weeks  2  3  4  197 196 201 213 206 220  269 264  380 394 400 449 406 466 348 353 353 377 355 368  178  177  189  194  181 178  F - female. M - male.  Table Feed  R a t i o n No.  1 2 3 4 5 6  11  Efficiency Feed e f f i c i e n c y f o r test period  2.32 2.11 2.19 3.03 2.73 2.73  28?  311 292 329 250 255 255 268 257 259  / - 100 -  FINAL AVERAGE WEIGHTS AT 4 WEEKS  1. BASAL-STARCH  F  1. BASAL-STARCH-  M  2. #1 f ,0.5% ANTIB *  F  2. #1 t,°.5% ANTIB.  M  3. #1 f 1.0% ANTIB.  F  3. #1 f 1.0% ANTIB.  M  4. BASAL-ALPHACEL *  F  4. BASAL-ALPHACEL- *  M  5. #4 f 0.5% ANTIB.  F  5. #4 + 0.5% ANTIB.  M  6.  F  #4 ¥ 1.0% ANTIB-.  6.^ #4 4- 1.0% ANTIB-.  M 350  400  450  GRAMS'  F i g u r e 1, Average weights of male and female c h i c k s f e d r a t i o n s a t two carbohydrate l e v e l s supplemented w i t h two l e v e l s o f a n t i b i o t i c s .  *  Ground c e l l u l o s e .  - 101 -  IV.- DISCUSSION  A series the  o f nine t e s t s was designed t o demonstrate  e f f e c t of a n t i b i o t i c s f e d at d i f f e r e n t  different  chicle r a t i o n s .  l e v e l s and w i t h ,  A b r i e f summary o f the d i f f e r e n t  experiments i s as f o l l o w s : Experiment #1. The  first  experiment was designed t o show the e f f e c t  of a n t i b i o t i c s on growth o f c h i c k s when added t o a C o n n e c t i c u t ration,  a soybean r a t i o n and a f i s h m e a l r a t i o n and t o make a  comparison o f the growth response t o each r a t i o n . the  In  addition,  e f f e c t o f adding v i t a m i n s and m i n e r a l s t o these b a s a l  rations  was s t u d i e d both w i t h and without a n t i b i o t i c s .  Experiment §2. T e s t s 2, 3 and 4 i n v o l v e d a study o f the e f f e c t of a n t i b i o t i c s on the growth r a t e o f c h i c k s f e d r a t i o n s ing d i f f e r e n t  levels  of protein.  contain-  The study was designed t o  determine whether a n t i b i o t i c s had any e f f e c t on l o w e r i n g the protein  needs o f the c h i c k .  different any  A t e s t was a l s o  conducted on  l e v e l s o f a n t i b i o t i c s i n the r a t i o n s  difference  t o determine  i n growth response.  Experiment #3. T e s t s 5,' 6 and 7 i n v o l v e d a study t o show the e f f e c t of a n t i b i o t i c s on r a t e o f growth o f c h i c k s when.used i n '  - 102 -  rations  i n which s p e c i f i c amino a c i d s were d e f i c i e n t .  o b j e c t o f these t e s t s ,  t h e r e f o r e , was t o determine i f  a n t i b i o t i c s had any e f f e c t on lowering the d i e t a r y ments f o r these l i m i t i n g amino a c i d s . made w i t h d i f f e r e n t difference  levels  the  require-  T e s t s , again, were  of a n t i b i o t i c s t o determine any  i n r a t e o f growth a t the l e v e l s used.  of adding a d d i t i o n a l  The  v i t a m i n s was a l s o  The e f f e c t  determined i n some o f  rations. Experiment #4. T e s t s 8 and 9 were designed t o study the e f f e c t o f  a n t i b i o t i c s on r a t e o f growth o f c h i c k s f e d r a t i o n s different  levels of available  carbohydrates.  containing  The o b j e c t was  to determine whether o r not a n t i b i o t i c s had any e f f e c t on lowering the requirement of the c h i c k f o r carbohydrate. The the  addition  results  from the f i r s t  experiment showed t h a t  o f a n t i b i o t i c s t o each o f the r a t i o n s  produced a s i g n i f i c a n t growth i n c r e a s e .  described,  The h i g h e s t growth  i n c r e a s e was obtained w i t h the C o n n e c t i c u t r a t i o n when a n t i b i o t i c s were f e d .  The C o n n e c t i c u t and soybean  supplemented w i t h v i t a m i n s and m i n e r a l s produced no  rations additional  growth whereas the f i s h m e a l r a t i o n when thus supplemented d i d produce a d d i t i o n a l  growth.  Similar  r e s u l t s were o b t a i n e d  when a n t i b i o t i c s were f e d i n these r a t i o n s . example, i s known t o be d e f i c i e n t  Fishmeal, f o r  i n f o l i c acid  (65) which  - 103 -  may be the reason f o r the added growth. g r e a t e s t response  The f a c t t h a t the  t o a n t i b i o t i c was gained w i t h the C o n n e c t i c u t  r a t i o n i s i n agreement with work done by Berg w i t h a h i g h energy r a t i o n .  (8) working  When he s u b s t i t u t e d the h e r r i n g  meal i n the r a t i o n and added more soybean o i l meal the growth response was  lowered.  These r e s u l t s seem t o i n d i c a t e t h a t the response t o a n t i b i o t i c s i s dependent, i n p a r t at l e a s t , on the composition of the r a t i o n w i t h which i t i s f e d .  T h i s _ i s v e r i f i e d by work  done by S c o t t e t a l (18) i n t e s t i n g an a l l - v e g e t a b l e r a t i o n c o n t a i n i n g ground corn, soybean o i l meal, crude soybean o i l , dl-methionine and m i n e r a l s p l u s the known v i t a m i n s .  They  found t h a t only supplements i n c l u d i n g corn d i s t i l l e r s ' s o l u b l e s , and b u t y l f e r m e n t a t i o n at the %  l e v e l and a l s o 2%  of a grass j u i c e f a c t o r concentrate gave a d e f i n i t e growth response  i n the t e s t r a t i o n when aureomycin was i n c l u d e d .  Other supplements such as dehydrated  a l f a l f a meal, d e f a t t e d  l i v e r meal, d r i e d brewers' y e a s t , and d r i e d c e r e a l grass ( r y e ) had no e f f e c t when aureomycin was added.  McGinnis  (58) has  r e p o r t e d t h a t f i s h m e a l , l i v e r L - f r a c t i o n o r d r i e d whey produced  g r e a t e r growth response w i t h c h i c k s o r p o u l t s i n  the presence  o f a n t i b i o t i c s than was the case when a n t i b i o t i c s  were not added and a l s o t h a t a n t i b i o t i c s produced  a greater  response when added t o r a t i o n s c o n t a i n i n g these animal sources than when added t o an a l l - v e g e t a b l e type  protein  ration.  - 104 -  At l e a s t one a d d i t o n a l u n i d e n t i f i e d f a c t o r , present i n f i s h m e a l , -fish s o l u b l e s , c e r t a i n l i v e r f r a c t i o n s and d r i e d whey, was r e q u i r e d poults.  f o r maximum growth o f c h i c k s and t u r k e y  Other r e p o r t s  (19, 38) showed t h a t the presence of  an animal p r o t e i n supplement i n a r a t i o n improved the growth response when a n t i b i o t i c s were i n c l u d e d The  i n the r a t i o n .  second s e r i e s of t e s t s (Experiment 11) on the  e f f e c t o f a n t i b i o t i c s on c h i c k s  f e d diets containing d i f f e r e n t  l e v e l s o f p r o t e i n showed t h a t the a n t i b i o t i c s had no e f f e c t on l o w e r i n g the p r o t e i n needs of the c h i c k .  The b a s a l  rations  used were composed o f equal p a r t s o f wheat and cornmeal, equal parts o f soybean o i l meal and f i s h m e a l ingredients  t o balance the r a t i o n s .  along w i t h a d d i t i o n a l  The p r o t e i n l e v e l s were  adjusted  by a l t e r i n g the r e l a t i v e amounts o f soybean and  fishmeal  t o the corn and wheatmeal.  contrast to reports containing  These r e s u l t s are i n  by Berg (38) i n comparing r a t i o n s  d i f f e r e n t l e v e l s of protein.  I n three  trials i t  was found that 19% p r o t e i n promoted as good growth as 21% protein.  The r e s u l t s , however, are  i n agreement w i t h work done  by McGinnis (39) on t u r k e y s when f e d p e n i c i l l i n rations at different protein l e v e l s . was kept a t a constant l e v e l ,  i n practical  The f i s h m e a l ,  and the p r o t e i n l e v e l s  however, adjusted  by changing the r e l a t i v e amounts o f soybean o i l meal and ground corn.  He found t h a t the a d d i t i o n o f p e n i c i l l i n d i d  not lower the requirement o f the turkey p o u l t f o r p r o t e i n .  - 105 -  M a c h l i n et a l (40)  on the o t h e r hand found t h a t the p r o t e i n  l e v e l i n c h i c k r a t i o n s c o u l d he lowered from 21 t o 19 per cent. They used a corn-soybean r a t i o n and reduced the corn and soybean meal i n c o n s t a n t p r o p o r t i o n s t o reduce the p r o t e i n level.  The r e d u c t i o n s were r e p l a c e d w i t h g l u c o s e ( c e r e l o s e ) . An aureomycin supplement  aureomycin HC1  levels  (1.8  gms.  per 100 l b . r a t i o n ) r e s u l t e d i n no added  growth over 0.5% ration).  f e d a t 1.0%  l e v e l (0.9  gms.  aureomycin HC1  McGinnis (21) found t h a t 0.1 gm.  100 l b . r a t i o n was  per 100 l b .  of aureomycin per  the minimum amount t o be used i n r a t i o n s .  Kramke and F r i t z (23)  r e p o r t e d t h a t 0.5  gm.  per 100 l b . r a t i o n  of aureomycin gave optimum growth. In the t h i r d s e r i e s of t e s t s (Experiment 111) a n t i b i o t i c s and amino a c i d s , i t was  found t h a t  on  antibiotics  had no e f f e c t i n l o w e r i n g the needs of the c h i c k f o r s p e c i f i c amino a c i d s .  T h i s r e s u l t was  shown i n a wheat r a t i o n  d e f i c i e n t i n l y s i n e t o which an a n t i b i o t i c was  added and a l s o  i n a corn r a t i o n d e f i c i e n t i n t r y p t o p h a n t o which an a n t i b i o t i c was  a g a i n added.  (1.5 gm.  No d i f f e r e n c e was  found between p e n i c i l l i n  per 100 l b . r a t i o n ) and aureomycin (1.8  aureomycin HC1  gm.  per 100 l b . r a t i o n ) i n t h e i r growth promoting a  a c t i v i t y when used i n these r a t i o n s .  Jones and Combs (41)  found t h a t aureomycin appeared t o spare the d i e t a r y r e q u i r e ment f o r tryptophan but not f o r l y s i n e .  They f e d p r a c t i c a l  type r a t i o n s to c h i c k s which were suboptimal i n l y s i n e  and  - 106  tryptophan.  -  Sherwood and Couch (37) found aureomycin  not lower the requirements i n a r a t i o n supplemented cottonseed meal as a p r o t e i n .  T h i s was  did  with  again v e r i f i e d  by  M a c h l i n et a l (42) w h i l e S l i n g e r et a l (43) found s i m i l a r r e s u l t s i n s t u d y i n g l y s i n e d e f i c i e n c i e s and a n t i b i o t i c s i n r e l a t i o n t o white f e a t h e r s i n t u r k e y s .  They suggested  that  a n t i b i o t i c s , m a y even i n c r e a s e the requirement of t u r k e y s f o r l y s i n e and perhaps  o t h e r amino a c i d s .  and aureomycin McGinnis  In comparing  penicillin  (21) found t h a t procaine p e n i c i l l i n  appeared t o be more e f f e c t i v e than aureomycin c h i c k and t u r k e y growth.  B r a n i o n and H i l l  i n promoting .  (20) found t h a t  turkeys on an animal p r o t e i n d i e t at 8 weeks of age gave the g r e a t e s t growth response t o aureomycin  and i n the case of a  p l a n t p r o t e i n d i e t at the same age a g r e a t e r response t o penicillin.  However, at f o u r weeks of age, p e n i c i l l i n  the g r e a t e s t growth response w i t h both In  gave  diets.  the f o u r t h s e r i e s of t e s t s i t was  found that  a n t i b i o t i c s had no e f f e c t i n l o w e r i n g the carbohydrate requirement of the c h i c k . soybean  Again,ground  corn and wheat and  o i l meal and f i s h m e a l were the main i n g r e d i e n t s  used along w i t h added i n g r e d i e n t s t o balance the r a t i o n . The a v a i l a b l e carbohydrate l e v e l was. lowered by s u b s t i t u t i n g 15% a l p h a c e l f o r a p a r t of the wheat and c o r n meal i n the . ration.  S t a r c h (15%) was  used i n p l a c e of the a l p h a c e l t o  r e s t o r e the a v a i l a b l e carbohydrate i n the b a s a l r a t i o n .  - 107 The  r e s u l t s a l s o showed t h a t aureomycin, when f e d at  d i f f e r e n t l e v e l s ( 0 . 5 and requirement of the  1.0%),  d i d not lower the  two carbohydrate  chick.  Considerable  evidence  has  been found i n the  l i t e r a t u r e t o i n d i c a t e t h a t there i s a - r e l a t i o n s h i p between A t k i n s o n and Gouch (25)  a n t i b i o t i c s and the v i t a m i n s . t h a t a n t i b i o t i c s were necessary  showed  i n a l l - v e g e t a b l e rations, to  promote growth when p o u l t s were i n j e c t e d with v i t a m i n Bi2« The  i n j e c t i o n of B]_2 alone had no e f f e c t on growth r a t e .  Reynolds, et a l , (26)  found t h a t f e e d i n g t e r r a m y c i n  commercial r a t i o n s reduced the requirement f o r B12. found t h a t even as t e r r a m y c i n had a B12  v i t a m i n B12  found aureomycin was  i n the r a t maintained  p o s t u l a t e d t h a t aureomycin may  They  sparing effect,  v i t a m i n a l s o showed an a n t i b i o t i c s p a r i n g e f f e c t . ffiunoz, et a l , (27)  i n good  the  Craviato-  able t o r e p l a c e  on a s y n t h e t i c d i e t .  They  decrease the number of  E s c h e r i c h i a c o l i i n the i n t e s t i n a l t r a c t and a l l o w the i n c r e a s e of b a c t e r i a such as B a c i l l u s megatherium which s y n t h e s i z e adequate v i t a m i n B12 and Jukes (29)  f o r r a t growth.  Stokstad  found t h a t aureomycin spared v i t a m i n B12  some cases but not i n o t h e r s .  can  in  They found t h a t aureomycin  was  e f f e c t i v e i n lowering m o r t a l i t y e s p e c i a l l y i n the absence of v i t a m i n B12.  I t was  suggested t h a t aureomycin may  s p a r i n g a c t i o n on v i t a m i n B12  have a  or reduce the number of  c e r t a i n microorganisms i n the i n t e s t i n a l t r a c t which are  - 108 -  i n j u r i o u s t o the b i r d .  L i n k s w i l e r , Baumann and S n e l l (31)  found t h a t aureomycin improved the growth o f r a t s when f e d the three forms o f v i t a m i n B6 ( p y r i d o x a l , pyridoxamine and pyridoxine)  whereas p y r i d o x i n e  the absence o f t h i s a n t i b i o t i c .  was the only e f f e c t i v e form i n L i h and Baumann (32) found  t h a t a n t i b i o t i c s s t i m u l a t e d growth i n young r a t s r e c e i v i n g l i m i t i n g amounts o f thiamine, r i b o f l a v i n o r pantothenic  acid.  B i e l y and March (17) found t h a t aureomycin f e d t o c h i c k s lowered the d i e t a r y requirement f o r f o l i c a c i d , n i c o t i n i c a c i d and  riboflavin.  A r e p o r t by Sunde, e t a l , (33) showed t h a t  one  f u n c t i o n o f a n t i b i o t i c s was the s p a r i n g o f v i t a m i n B]_2  and  the i n d i r e c t s p a r i n g o f methionine and c h o l i n e  requirements  of the c h i c k . The  r e s u l t s from experiment 1 i n d i c a t e d t h a t the  a d d i t i o n o f aureomycin improved the f e e d e f f i c i e n c y when f e d i n the Gonnecticut, soybean and f i s h m e a l  rations.  Tests i n  experiment 11,however, showed t h a t the f e e d e f f i c i e n c y was lowered when a n t i b i o t i c s were f e d i n r a t i o n s with sub-optimal protein levels.  S i m i l a r r e s u l t s were obtained  i n the t e s t s  i n experiment 111 when amino a c i d s were d e f i c i e n t and i n experiment IV when the carbohydrate l e v e l was lowered. and March (44) showed t h a t aureomycin i n c r e a s e d feed  Biely  efficiency  i n a 17 per cent p r o t e i n d i e t c o n t a i n i n g f i s h meal over a 21 per cent p r o t e i n a l l - v e g e t a b l e r a t i o n c o n t a i n i n g Some r e p o r t s  aureomycin.  (18, 15, 45, 49) found t h a t a n t i b i o t i c s d i d not  - 109 improve feed e f f i c i e n c y when f e d i n d i f f e r e n t other r e p o r t s (40, in  feed  46,  47,  found a d e f i n i t e  action  of a n t i b i o t i c s has  d e f i n i t e l y established. indicate  improvement  the  Reports (17,  s t i l l not  27,  28,  s p a r i n g e f f e c t of a n t i b i o t i c s  suggest that f e e d i n g an a n t i b i o t i c may  b a c t e r i a l count i n the  intestine,  There i s a l s o  might i n h i b i t c e r t a i n  the  30,  vitamins  decrease  the  animal from  p e r m i t t i n g more  antibiotics allow a  type to f l o u r i s h which s y n t h e s i z e s a growth f a c t o r .  Speer,  and  S i e b u r t h , et a l , (60)  the  a n t i b i o t i c s act by  and  thus prevent a r e t a r d a t i o n  intestine  rapid  and  et a l , (14)  i n the  32)  on the B  p o s s i b i l i t y t h a t the  bacteria  been  29,  r e l i e v i n g the  c o m p e t i t i o n f o r these v i t a m i n s and growth.  whereas  efficiency. The  and  48)  rations,  support the  inhibiting toxin  producing  aureomycin f e e d i n g lowered the  may  be  S h a p i r o and  i n the  shown t h a t  L a c t o b a c i l l i i n the  S i n c e these b a c t e r i a  a factor  bacteria  of growth.  A study by B i e l y and March ( 6 l ) has  of c h i c k s .  view t h a t  are  intestine  v i t a m i n consuming, t h i s  growth s t i m u l a t i n g e f f e c t of aureomycin.  S a r l e s (59)  have shown t h a t L a c t o b a c i l l i are  most numerous group of b a c t e r i a  i n most areas of the  the  intestinal  t r a c t of normal b i r d s . There i s s t i l l i n g the the  a great d e a l to be  action  of a n t i b i o t i c s  intestine.  Much i n t e r e s t  on the  l e a r n e d concern-  microbe p o p u l a t i o n  i s being shown at the  of  present  - 110-  time i n the r e s u l t s being  obtained  by R e y n i e r s and h i s  a s s o c i a t e s a t the U n i v e r s i t y of Notre Dame i n r e s e a r c h germ-free l i f e  i n chicks.  C h i c k s can be r e a r e d  on  i n special  equipment a b s o l u t e l y germ-free and develop as w e l l as c o n t r o l chicks on the same d i e t .  Perhaps more work o f t h i s nature  r e s u l t i n g i n more fundamental i n f o r m a t i o n w i l l r e v e a l the mode o f a c t i o n o f a n t i b i o t i c s .  - Ill -  V. SUMMARY  A study has been made on the growth stimulatory e f f e c t of a n t i b i o t i c s when fed to chicks as supplements to d i f f e r e n t rations. A t o t a l of 9 b i o l o g i c a l tests employing1280 chicks were c a r r i e d out. The r e s u l t s obtained are summarized as follows: 1.  Chicks fed the Connecticut r a t i o n and modifications thereof showed a s i g n i f i c a n t growth response to aureomycin supplementation of the rations. 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 improved by the addition of aureomycin to the rations i n t h i s experiment.  2.  The e f f e c t of aureomycin has been tested i n rations i n which the protein l e v e l has been adjusted to 16%, 17.9$, 19* 5% and 22% respectively. The basal rations used consisted of ground corn and wheat as the cereal grain and soybean o i l meal and fishmeal as the protein supplements along with a d d i t i o n a l vitamins and minerals to balance the r a t i o n . The r e s u l t s showed that aureomyoin did not lower the dietary requirement of the ohiok f o r protein. 1  3.  A test was made on the e f f e c t of a n t i b i o t i c s on growth of chicks when f e d i n rations which were d e f i c i e n t i n s p e c i f i c amino acids. A wheat r a t i o n d e f i c i e n t i n lysine and a corn r a t i o n d e f i c i e n t i n tryptophan have been used. The results showed that a n t i b i o t i c s did not lower the dietary requirement of the chick f o r e i t h e r l y s i n e or tryptophan.  4.  A n t i b i o t i c s were f e d In chick rations containing d i f f e r e n t amounts of a v a i l a b l e carbohydrates. I t was found that a n t i b i o t i c s did not lower the dietary requirement of the chick f o r carbohydrate.  5.  A comparison of aureomycin and p e n i c i l l i n showed no difference i n growth stimulating properties between the two a n t i b i o t i o s under the conditions of the d i f f e r e n t experiments. Aureomycin supplement (aurofac) promoted the same growth-response at 0.05 and 1.0% l e v e l s .  6.  The greatest growth response to a n t i b i o t i c s was obtained when the chicks were fed a diet adequate i n available carbohydrates and protein and balanced i n amino acids.  -  112 -  BIBLIOGRAPHY  1.  Moore, P.R., A. Evens on, T.D. Luckey, E. McCoy, C A . Elvehjem, and E.Bs. Hart, 1946, "Use of sulfasuxidine, streptothracine and streptomycin i n n u t r i t i o n a l studies with the chiok." Journ. B i o l . Chem., :  165:  437-441.  2.  Stokstad, E.R.R., T.H. Jukes, F. Pierce, A.C. Page, and A.L. Franklin, 1949. "The multiple nature of A.P.F." Journ. B i o l . Ohem., 180: 647.  3.  Cunha, T.J., J.E. Burnside, D.M. Buschman, R.S. Glasscock, A.M. Pearson, and A.L. Shealy, 1949. Arohiv. Biochem., 23:  324.  4.  Catron, D.V., 1950. Feedstuffs. June 24: 46.  5.  Carpenter, R.S., 1950. The Hormet Farmer. 13: A p r i l : 4 - 7 .  6.  McGinnis, J . , E.L. Stephenson, B.T. Levadie, J.S. Carver, J.A. Garabaldi, N.S. S n e l l , J.G. Lewis, 1950. "Response of chicks and turkey poults to vitamin B j ^ supplements produced by fermentation with d i f f e r e n t organisms." Abstracts of paper, 116th meeting of Amer. Chem. Soo: 42a.  7.  Singsen, E.P., and L.D. Matterson, 1949. New England Feedmans Meeting. Conn. Agric. Exp. Stn., August: 18.  8.  Berg, R.R., G.E. Bearse, J . McGinnis and V.L. M i l l e r , 1950. " E f f e c t of growth of f r y e r s of adding aureomycin fermentation produet to a high energy r a t i o n . " Poultry Science. 29: 921.  9.  Stokstad, E.R.R. and T.H. Jukes, 1950. "Further studies on the Animal Protein Factor." Soc. Exp. B i o l , and Med.. 73: 523.  10.  Whitehall, A.R., J . J . Olson, B.L. Hutohings, 1950. "Stimulatory e f f e c t of aureomycin on the growth of chicks." Proo. Soo. B i o l , and Med.. 74: 11-13.  r 113 -  11.  Leuke, R.W., W.N. McMillan, F. Thorp, 1950. "The e f f e c t of vitamin B12, Animal Protein Factor and Streptomycin on the growth of young p i g s . Archiv. Biochem., 2 6 : 326-327. n  12.  McGinnis, J . . 1950* "Vitamin B12 and second growth f a c t o r (s) i n A.P.F. supplement." Wash. State College N u t r i t i o n Conference, A p r i l 3 - 4 : 45.  13.  Davies, R.L. and G.M. Briggs, 1951. "Studies with A n t i b i o t i c s i n Chick and Poult S t a r t i n g Rations." Poultry Science. 3 0 : 767-771. Speer,V.C., R.L. Vokes, D.V. Catron, H.M. Haddock, and  14. 15.  C.G. Cuthbertson, 1950. Archives Biochem. 2 9 : 4 5 2 . Scott, H.M., W.A. G l i s t a , 1950. "The e f f e c t of aureomycin and streptomycin on growth of chicks and turkey poults." Poultry Science. 29.: 7 7 1 .  16.  Oleson, J.J"., B.L. Hutchings, and A.R. W h i t e h i l l , 1950. Archiv. Bioohem.. 2 9 : 3 3 4 .  17.  B i e l y , J . , and B. Maroh, 1 9 5 1 . "The e f f e c t of aureomycin and vitamins on the growth rate of chicks." Science, 114:. 330-331. ~  18.  Scott, H.M., W.A. G l i s t a and E.A. G a f f i , 1 9 5 1 . "Growth effects of oertain supplements added to a cornsoybean o i l meal chick r a t i o n with and without a n t i b i o t i c . " Poultry Science. 3 0 : 9 3 0 .  19.  Matterson, R.D., et a l . 1 9 5 1 . "A comparison of several a n t i b i o t i c s as growth stimulants i n p r a c t i c a l chick . s t a r t i n g r a t i o n s . " Conn. Storrs* Ag. Exp. B u l l e t i n . 275: 2 0 .  20.  Branion, H.D., and D.C. H i l l , 1951. "The comparative effect of a n t i b i o t i c s on the growth of poults." Poultry Science. 3 0 : 7 9 3 .  21.  McGinnis, J . , 1 9 5 1 . " A n t i b i o t i c s i n feeds f o r chicks and poults." 'Wash. State College N u t r i t i o n Conference. A p r i l 2-3: 6 9 .  22.  McGinnis, J . , 1 9 5 1 . " A n t i b i o t i c s and poult growth." Wash. State College N u t r i t i o n Conference. Dec. 4 : 5 3 .  - 114  -  23.  Kromke, E.H., and J.C. F r i t z , 1951. "The response of chicks and poults to.low l e v e l s of a n t i b i o t i c s . " Poultry Science. 30: 921.  24.1  Reynolds, J.W., T.D. Runnels and E.F. Walles, 1951* "A comparison of terramycin and p e n i c i l l i n at various l e v e l s on rate of growth and feed e f f i c i e n c y of b r o i l e r d i e t s . " Poultry Science. 30: 928.  25.  Atkinson, R.R., and J.R. Couch, 1951. "Vitamin B12> an A.P.F. concentrate, aureomycin, streptomycin, l i v e r "L", and f i s h meal and f i s h solubles i n the n u t r i t i o n of the poult." J . of N u t r i t i o n . 44: 249-261.  26.  Reynolds, W.M., H.G. Luther and E.M. Feedstuffs. Aug. 4: 43.  27.  Cravioto-Munoz, J . , H.G. Poncher, and Harry A. Waisman, 1951. "Vitamin B12 sparing action of aureomycin i n the r a t . " Proc. Soc. B i o l , and Med.. 77: 18-19.  28.  Davis, B.D., and E.S. M i n g i o l i , 1950. "Mutants of Escherichia c o l i requiring methionine or vitamin B i 2 » "  d  J . of Bact.. 60:  29.  17-28.  Stokstad, E.L.R., and T.J. Jukes, 1951. " E f f e c t of various l e v e l s of vitamin B 1 2 upon growth response produced by aureomycin i n chicks." Proc. S00. B i o l . and  30.  Weber, 1951. *  Med..  76:  27-30.  Davis, L.D., and F.G. Bacon, 1951. The extent of radioactive vitamin B12 i *he feces of rats fed CoAo and aureomycin." Proc. Soc. B i o l , and Med.. rt  n  77:  31.  ~~  LinkswiIer, H.M., C A . Maumann, and E.E. S n e l l , 1951. " E f f e c t of aureomycin on growth response of rats to various forms of vitamin BA." J . of N u t r i t i o n . 43:  32.  218-221.  565.  L i h , Hwa, and C.A. Baumann, 1951. "Effects of c e r t a i n a n t i b i o t i c s on the growth of rats fed diets l i m i t i n g i n thiamine, r i b o f l a v i n , or pantothenic a c i d . " J . of N u t r i t i o n . 45: 143.  33. - Sunde, W.L.,-Paul E-. Waibil, W.W. Cravens, and C.A. Elvehjem, 1951. "A r e l a t i o n s h i p between antibiotics,vitamin B12 choline and methionine i n chick growth." Poultry Science. 30: 668-671.  - 115 -  34. . Coates, M.E., C D . Dickinson, G.F. Harrison, and.S.K. Kon, 1 9 5 1 . "The e f f e c t of a n t i b i o t i c s on the growth of ohieks deprived of vitamins of the B complex.7 Biochem. Journal. 4 9 : 9 8 . 35.  Burgess, R.C., et a l . 1 9 5 1 . " E f f e c t of dietary p e n i e i l l i n on l i v e r vitamin"A and serum carotenoids i n the chick." Arch, of Biochem. and Physios. 3 3 : 3 3 9 .  36.  Burnside, J.E., T.J. Cunha, A.M. Pearson, E.S. Glassock, A.L. Shealy, 1950. " E f f e c t of A.P.F. supplements on pigs f e d d i f f e r e n t protein supplements." Arch, of Biochem.. 2 3 : 328-330.  37.  Sherwood, R.M., and J.R. Gouch, 1950. "The effect on chick growth of supplementing a vegetable protein diet with an A.P.F. concentrate." Poultry Science. 29:.501-507.  38.  Berg, L.R., 1 9 5 1 .  ^ A n t i b i o t i c s i n feeds f o r b r o i i e r s . "  Wash. State College N u t r i t i o n Conference. A p r i l 2-3: 65. 39. 40.  McGinnis, J . , 1 9 5 1 . " A n t i b i o t i c s i n turkey n u t r i t i o n . " Wash. State College N u t r i t i o n Conference. Dec. 4 : 4 9 . Machlin, L.J., C.A. Denton, W.L. Kellogg, and H.L. B i r d , 1952. " E f f e c t of dietary a n t i b i o t i c upon feed e f f i c i e n c y and protein requirement of growing chicks." Poultry Science. 3 1 : 106-109.  41.  Jones, H.J., and G.F. Combs, 1 9 5 1 . " E f f e c t of a n t i b i o t i c s on the.dietary requirements f o r methionine, l y s i n e , tryptophan, and u n i d e n t i f i e d growth f a c t o r s . " Poultry Science. 3 0 : 9 2 0 .  42.  Machlin, L.J., C.A. Denton and H.R. B i r d , 1 9 5 2 . "Supplementation with vitamin Bi2» amino acids of chick diets containing soybean and cottonseed meal." Poultry Soience. 3 1 : 110-114. 6111(1  43.  Slinger, S.J.-, K.M. Gartby, W.F. Pepper, and D.C. H i l l , 1951. "The influence of animal protein supplements and a n t i b i o t i c s on the Incidence and severity of white feathers i n turkeys." J . of N u t r i t i o n . 4 3 : 345-366, , . ' ; .  - 116 44*  Biely,., J., B. March, Joan Stevens, and E . Casorso, 1951* "A.P.F.. supplements i n the chick ration.". Poultry. Science. 30:.143-147.  45.  Groschke, A.G., R.J. Evans, 1950. " E f f e c t of a n t i b i o t i c s , synthetic vitamins, vitamin Big and A.P.F. supplement on chick growth." Poultry Science. 29: 616-618.  46.  Atkinson, R.L., and J.R. Couch, 1951. "The e f f e c t of feeding an A.P.F. concentrate and f i s h solubles on the growth of broad breasted bronze turkeys." Poultry Science. 30: 81-85.  47.  Bowland, J.P., J.E. Beacon, and L.W. McElroy, 1950. "Animal protein f a c t o r supplementation of small grain rations for. swine." J. of Animal S o i . . 9: 649.  48.  Carpenter, L.E., 1951. " E f f e c t of aureomycin on the growth of weaned pigs." Arch, of Biochem.. 2 7 :  469-471.  49.  Beeson, M., 1950. "The supplementary effect, of B vitamins, A.P.F. concentrates-^ and streptomycin on growth of weanling pigs." . J . of Animal Sol... 9: 649.  50. >a  H i l l , F.W., 1948. ''The multiple nature of the deficiency of u n i d e n t i f i e d nutrients i n crude all^vegetable protein chick s t a r t e r rations." Poultry Science,  27:  536-541.  51.  Carlson, CW., R.F. M i l l e r , H.T. Peeler, L . C Norris, and, G.F.- Heuser, 1949. "The complex nature of the animal protein faetor." Poultry Science. 28: 750-752.  52.  Wiese, A . C , G.F. Peterson, and C E . Lampman, 1949. " A c t i v i t y of a microbial animal protein factor concentrate in. a< high energy b r o i l e r ration.!* Poultry Science. 28: 752-754.  53.  Sunde, M.L., W.W. Cravens, C A . Elvehjem,. and J.G. Halpin, 1950. "An u n i d e n t i f i e d faetor required by chicks fed p r a c t i c a l rations.! Poultry Science. 1  29:  54.  204-207.  H i l l , D.C, and H.D. Branion, 1950. "A .comparison of commercial animal protein f a c t o r supplements i n chick s t a r t e r d i e t s . " Poultry Science. 29: 743-744.  - 117  -  55.  Berg, L.R., G.E. Bearse, J . McGinnis, and V.L. M i l l e r , 1950. "The e f f e c t of removing supplemental aureomycin from the r a t i o n on the subsequent growth of chicks." Arch, of Biochem.. 2 9 : 404-407.  56.  McGinnis, J . , J.R. Stern, R.A. Wilcox, and J.S. Carver, 1 9 5 1 . "The e f f e c t of d i f f e r e n t a n t i b i o t i c s on growth of turkey poults." Poultry Science, 30: 492-496.  57.  McGinnis, J . , L.R. Berg, J.R, Stern, M.E. S t a r r , R.A. Wilcox, and J.S. Carver, 1952. "Additional evidence f o r an u n i d e n t i f i e d growth promoting f a c t o r f o r turkeys and chicks." Poultry Science, 3 1 : 100-106.  58.  McGinnis, J . , 1951. Feedstuffs. 23:  59.  Shapiro, S.K., and W.B. Sarles, 1949. "Microorganisms i n the i n t e s t i n a l t r a c t of normal chickens.? J . of Bact.. 58: 531-544.  60.  Sieburth, J.M., J . Gutierrez, J . McGinnis, J.R. Stern, B.H. Schneider, 1951. " E f f e c t of a n t i b i o t i c s on i n t e s t i n a l microflora and growth of turkeys and pigs." Proc. Soc. Exper. B i o l , and Med.. 76: 15-16.  61.  March, B., and J . B i e l y , 1952. HThe e f f e c t of feeding aureomycin on the b a c t e r i a l content of chick feces." Poultry Science. 3 1 : 1 7 7 .  62.  Dixon, J . , and R.H. Thayer, 1952. "Observations on the growth promoting action of a n t i b i o t i c s i n the chick." * Poultry Science. 3 1 : 910-911.  63.  Anderson, G.W., J.D. Cunningham, and J.S. S l i n g e r , 1951. " E f f e c t of various protein l e v e l s and a n t i b i o t i c s on the i n t e s t i n a l f l o r a of chickens." Poultry Science. 30: 905.  64.  Kratzer, F.H., C.R. Grau, P Mortimer, and P.M. Reynolds, 1951. -"Growth promoting a c t i v i t b s of a n t i b i o t i c s and yeast cultures f o r chicks and turkey poults." Federation Proceedings. 1 0 : 285.  65.  L i l l i e , Robt. J . , and G.M. Briggs, 1946. " F o l i c acid requirements of New Hampshire chicks receiving synthetic d i e t s , " Poultry Science. 2 6 : 295.  "What's new 40.  i n poultry n u t r i t i o n . "  1  

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