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A biological study of the protein and riboflavin content of British Columbia fishmeals Pratt, Jean Margaret 1942

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A BIOLOGICAL STUDY OF THE PROTEIN AND RIBOFLAVIN CONTENT OF BRITISH COLUMBIA FISHMEALS  by J e a n M.  A Thesis submitted  Pratt  in Partial  Fulfilment  The R e q u i r e m e n t s f o r t h e D e g r e e o f MASTER OF SCIENCE IN AGRICULTURE  i n t h e Department of POULTRY HUSBANDRY  The U n i v e r s i t y o f B r i t i s h C o l u m b i a April,  1942  of  ACKNOWLEDGMENTS  The w r i t e r w i s h e s t o t a k e t h i s o p p o r t u n i t y t o t h a n k Mr. J a c o b B i e l y , o f t h e Department  of Poultry  f o r h i s g u i d a n c e and h e l p f u l s u g g e s t i o n s  Husbandry,  i n c a r r y i n g out  t h i s i n v e s t i g a t i o n ; M r • E . A. L l o y d , Head o f t h e D e p a r t ment o f P o u l t r y H u s b a n d r y ,  for h i s valuable  in the preparation of this thesis;  assistance  and Mr. C a r l  Hedreen,  o f t h e C a n a d i a n F i s h i n g Company, V a n c o u v e r , B C . , f o r e  s u p p l y i n g t h e f i s h m e a l s and b y - p r o d u c t s used i n the study.  t  •  if ' ••  .  '  • .  fj ;j  Table of Contents  i\  I  INTRODUCTION  1 " ||  i i i 1 I 4 I. ll  . ...  '  •U  "  1  . .  •"  .  •  " EISHMEALS 3 5 7 11 13 17 18  (a) Raw M a t e r i a l s ..«..••••••••»»»•«»••«»««««•• (b) Methods o f M a n u f a c t u r e » ( c ) P r o t e i n s ..,...»•••••••••©•••.••••••••••••• (d) V i t a m i n G (©) M i n s r a l s • ( f ) E f f e c t s o f Method o f M a n u f a c t u r e « (g) Uses o f F i sh. M e a l s • <>•••«•••••••••••••••••• • SERIES  1 19  (a) The E x p e r i m e n t , (b) M a t e r i a l s and M e t h o d s  1  ••  •  20  ••  (c) R e s u l t s E x p e r i m e n t 1 ............................ Experiment 2 . . . . . . . . . Experiment 3 • (d) D i s c u s s i o n ^ ..»»»•«•.. • •  23 24* 26  28 29  SERIES 2 (a) The E x p e r i m e n t  .••..••..«...•.•.*...«•«.>••  31  ••••••••••• ••••••••»•  31  (c) H @ s u l t s •«•••••••«•••«•«••«•••»••••••*•••••  32  (b) M a t e r i a l s anCL Methods  Exp ©r i meD.t 1 • • • » • • • « • • • • • • • • © • • • • • • • • • • « •  32  Exp6riitt©nt 2 • • • • • • « • • • • * • • • • • • • • • • • • • • • •  37  • •••••  38.  •••••••••••••••••••••••••••••••«  4-0  Exp©3?jLTiient (d) D i s c u s s i o n  3 •  STJMMARY •••*«•••••«• •  •  • • • •©•••• ••••••••• •  REFiJJrtEN CES ••••••••••• •»••••• •••••«•••••••••••••• •  4* 2 44  A BIOLOGICAL STUDY OF THE PROTEIN AND RIBOFLAVIN CONTENT OF B R I T I S H COLUMBIA FISHMEALS  INTRODUCTION The  p r o t e i n content  o f t h e common g r a i n s i s d e f i c i e n t  i n b o t h q u a n t i t y and q u a l i t y t o meet t h e n u t r i t i v e r e q u i r e ments o f g r o w i n g c h i c k s , l a y i n g b i r d s , and b r e e d i n g Hence, t h e p r o t e i n c o n t a i n e d  i n the grain i s generally  mented by t h e a d d i t i o n o f p r o t e i n c o n c e n t r a t e s . of animal  origin,  extent  o f vegetable  soyabean, l i n s e e d o r cottonseed protein concentrates  meals.  i stwofold:  the  such as  to increase the  1 0 - 1 2 / -  v i d e d by t h e g r a i n s o r g r a i n b y - p r o d u c t s t o second, t o complete and t o b a l a n c e  products,  The p u r p o s e o f t h e  first,  o f t h e r a t i o n from t h e  source,  supple-  These may be  s u c h a s f i s h m e a l , meat m e a l o r m i l k  o r t o a more l i m i t e d  p r o t e i n content  stock.  ordinarily  1 8 - 2 0 ^ ;  pro-  and  t h e amino a c i d c o n t e n t o f  ration.  The  e s t a b l i s h e d p r a c t i c e i n f e e d i n g c h i c k s has been,  u n t i l r e c e n t l y , t o r e p l a c e a c e r t a i n amount o f meat m e a l and f i s h m e a l w i t h d r i e d s k i m m i l k i n o r d e r t o d e r i v e t h e optimum growth response from t h e r a t i o n .  Asmundson and B i e l y , f o r  example ( ? ) , r e p o r t e d t h a t a c o m b i n a t i o n salmon a s t h e s o u r c e  o f d r i e d s k i m m i l k and  o f p r o t e i n was more e f f e c t i v e t h a n s a l m o n  a l o n e , and a t a t o t a l o f 7.5  a n d lof. seemed t o have a  s u p p l e m e n t a r y e f f e c t upon one a n o t h e r . Asmundson  ( 5 ) also reported  F u r t h e r m o r e , B i e l y and  t h a t when f i s h m e a l  constituted  7 , 5 / 0 o f t h e ' p r o t e i n o f t h e r a t i o n , t h e a d d i t i o n o f more t h a n 5/i skimmilk  d i d not r e s u l t i n i n c r e a s e d  growth.  At t h a t  the advantage o f f e e d i n g m i l k i n a d d i t i o n t o f i s h m e a l  time  was  s u p p o s e d t o be due t o t h e a d d i t i o n o f c e r t a i n amino  acids  f o u n d i n t h e m i l k and n o t i n t h e f i s h m e a l *  I t was  thought,  moreover, that t h e 5 f « l e v e l o f m i l k p r o v i d e d  a l l the extra  e s s e n t i a l amino a c i d s .  I t seems more l i k e l y now, h o w e v e r , t h a t  t h i s e x t r a g r o w t h r e s p o n s e was due t o t h e a d d i t i o n a l r i b o f l a v i n s u p p l i e d a t t h a t l e v e l by t h e m i l k ; and t h a t t h e amount o f vitamin G provided  a t t h e 51° l e v e l w o u l d  by t h e d r i e d s k i m m i l k  a p p e a r t o be a d e q u a t e f o r optimum g r o w t h .  The t y p e o f f i s h m e a l u s e d by B i e l y and Asmundson t h e i r e x p e r i m e n t s was a p p a r e n t l y f l a v i n content  (3,5) in  considerably lower i n r i b o -  than d r i e d skimmilk.'  More r e c e n t l y s u c h  i m p r o v e m e n t s have been made i n t h e method o f m a n u f a c t u r e as t o allow fishmeals t i v e value.  to r e t a i n a greater  percentage of t h e i r  nutri-  I t i s known, m o r e o v e r , t h a t due t o t h e c o n s t i t u -  t i o n o f f i s h m e a l s , and more p a r t i c u l a r l y t o t h e o r g a n s and tissues included  i n them, t h e y may v a r y  vitamin G content.  Since  to t h e r e l a t i v e potencies  little  considerably i n  i n f o r m a t i o n was a v a i l a b l e as  of d i f f e r e n t fishmeals  products of the f i s h i n g i n d u s t r y i n B r i t i s h  and b y -  C o l u m b i a , an  e x p e r i m e n t was begun i n t h e summer o f 1940 t o i n v e s t i g a t e t h e value  of these products.  Accordingly,  the f i r s t  series  i n v o l v e d a t e s t o f t h e e f f e c t s o f some o f t h e s e s u p p l e m e n t s t h e r a t e o f g r o w t h o f c h i c k s t o f i v e weeks o f age. second s e r i e s was  undertaken to i n v e s t i g a t e the  s u p p l e m e n t a r y v a l u e s o f s y n t h e t i c and n a t u r a l  on  Later a  relative  riboflavin.  FISHMEALS (a) Raw The  Materials i n c r e a s i n g use o f f i s h m e a l s as a p o u l t r y and  f e e d has been a c c o m p a n i e d  livestock  by a g r o w i n g r e c o g n i t i o n o f t h e  v a l u e o f m a r i n e p r o d u c t s as n a t u r a l s o u r c e s o f p r o t e i n o f a very high q u a l i t y .  There a r e , however, c o n s i d e r a b l e  varia-  t i o n s i n the u l t i m a t e value of the product, depending t h e n a t u r e o f t h e s p e c i e s o f f i s h f r o m w h i c h i t was and a l s o upon t h e method by w h i c h i t was  upon  prepared,  manufactured.  Since  t h e a d v e n t and e x p a n s i o n o f t h e k n o w l e d g e o f v i t a m i n s i t has been r e c o g n i z e d t h a t h e a t , l i g h t  and o x i d a t i o n may  bear a  c o n s i d e r a b l e i n f l u e n c e upon t h e f i n a l n u t r i t i v e v a l u e o f t h e product. The raw m a t e r i a l s f r o m w h i c h f i s h m e a l s a r e fall  i n t o two d i s t i n c t  manufactured  c l a s s i f i c a t i o n s , "based upon t h e  p h y s i o l o g i c a l c h a r a c t e r i s t i c s o f the f i s h from which the product o r i g i n a t e d  (19 ) •  The  first  c l a s s , the o i l y  i n c l u d e s the m a j o r i t y of those manufactured  fishmeals,  from f i s h  p i l c h a r d , t u n a , m a c k e r e l , menhaden and h e r r i n g ) w h i c h  (salmon, store  t h e f a t t h r o u g h o u t t h e t i s s u e s o f t h e body and have comparatively small l i v e r s .  The  second c l a s s , the n o n - o i l y meals  are prepared  f r o m f i s h w h i c h have l o c a l i z e d  l i v e r and o t h e r v i t a l o r g a n s . p o l l o c k , hake and content  cusk.  In t h i s  fat-depots i n the  c l a s s are cod,  This large v a r i a t i o n  i n the o i l  o f t h e raw m a t e r i a l s c a u s e s a c o r r e s p o n d i n g  i n the f i s h m e a l product.  Manning  haddock,  variation  published a bibliography  o f t h e c o m p a r i s o n s made between m e a l s p r e p a r e d  from  different  species of f i s h u n t i l that time, 1 9 3 0 . In  g e n e r a l , the f i s h m e a l s w i t h h i g h o i l content w i l l have  relatively  low p r o t e i n c o n t e n t , a l t h o u g h t h e r e a r e a  exceptions to t h i s statement.  However, t h e o i l has  few a  definite  v i t a m i n v a l u e w h i c h must be c o n s i d e r e d , s i n c e i t e n t e r s t h e r e d u c t i o n phase o f m a n u f a c t u r e . practice,"  "According to  into  existing  s t a t e H a r r i s o n et a l , ( 1 9 ) "the dehydrated  residue  a f t e r o i l r e m o v a l u s u a l l y c o n t a i n s 5 t o 15 p e r c e n t f a t , d e p e n d i n g upon t h e raw m a t e r i a l u s e d and t h e e f f i c i e n c y o f o i l e x t r a c t i o n process. r e d u c t i o n o f t h e two involved.  From t h i s  i t can be  the  s e e n t h a t i n the  types of waste, d i f f e r e n t  problems  are  I n t h e case o f t h e n o n o i l y w a s t e t h e r e i s t h e r e -  q u i r e m e n t o f d e h y d r a t i o n a l o n e , and  i n t h e case o f o i l y w a s t e  t h e r e i s t h e added p r o b l e m o f o i l r e m o v a l . "  In  a d d i t i o n to the lower p r o t e i n content, there i s  o b j e c t i o n to a high f a t content  i n fishmeals - namely  f a c t t h a t t h e y t e n d t o become r a n c i d t h r o u g h d e t e r i o r a t i o n upon s t a n d i n g .  t  another the  o x i d a t i o n and  E w i n g ( 1 5 ) s t a t e s t h a t i t has  been t h e e x p e r i e n c e o f t e c h n o l o g i s t s t h a t i n o r d e r f o r a f i s h m e a l t o r e m a i n a s t a b l e commodity d u r i n g s t o r a g e  and  - 5 handling,  i t should not contain  moisture. is liable the  more t h a n 6f° f a t o r 6f.  Excessive percentages of e i t h e r of these  factors  t o c a u s e a r a p i d d e c o m p o s i t i o n and d e t e r i o r a t i o n o f  n u t r i t i v e value,  a s w e l l a s t h e danger o f o v e r h e a t i n g .  A c c o r d i n g t o t h e F e e d i n g S t u f f s A c t o f Canada, 1937, it  i s provided that  fishmeal  o r any o t h e r p r o d u c t  (l6)  (except  l i v e r m e a l ) o f f i s h o r f i s h w a s t e be g u a r a n t e e d on a l a b e l as t o i t s minimum amount o f c r u d e p r o t e i n ; maximum amount o f c r u d e f a t ; maximum amount o f c r u d e f i b r e and  maximum amount o f s a l t  (NaCl).  i f i n excess o f 2y»;  F i s h l i v e r m e a l s need  show l a b e l s g u a r a n t e e i n g t h e minimum amounts o f c r u d e and  minimum and maximum amounts o f c r u d e f a t .  fishmeal  i s "the clean,  d r i e d , ground r e s i d u e ,  protein  By d e f i n i t i o n , c o n t a i n i n g not  more t h a n 6 p e r c e n t o f o i l , f r o m undecomposed w h o l e  fish  a n d / o r f i s h c u t t i n g s " , and O i l y F i s h M e a l i s " t h e c l e a n , ground r e s i d u e ,  containing  fishmeals  of the non-oily  cuttings"• class are generally  d e s i g n a t e d c o m m e r c i a l l y as w h i t e - f i s h meals, w h i l e the  dried,  more t h a n 6 p e r c e n t o f o i l , f r o m  undecomposed whole f i s h a n d / o r f i s h The  only  those of  o i l y c l a s s a r e known by t h e s o u r c e f r o m w h i c h t h e y were  p r e p a r e d -- a s Salmon m e a l , A l a s k a h e r r i n g m e a l , e t c . (b) Methods o f M a n u f a c t u r e T h e r e a r e s e v e r a l methods e m p l o y e d i n t h e p r e p a r a t i o n fishmeals,  explained  by D a n i e l  of  a n d M c C o l l u m (12) as f o l l o w s :  -  6.  -  "The raw m a t e r i a l s h o u l d be c l e a n and f r e s h i f i t i s t o be u t i l i z e d  f o r stock feeding.  cooked, pressed low.in o i l , are  T h i s raw m a t e r i a l i s steam  and d r i e d , o r i n t h e c a s e o f some f i s h m e a l s  s u c h as w h i t e  f i s h meals, the cooking  c a r r i e d o u t i n one o p e r a t i o n , t h e r e b y  i n g as a p a r t o f t h e p r o c e s s .  and d r y i n g  omitting the press-  The methods o f d r y i n g a f f o r d  the g r e a t e s t d i f f e r e n c e s i n the manufacturing  process.  e x a m p l e , d r y i n g may be a c c o m p l i s h e d i n r o t a r y d r y e r s  For  which  s u b j e c t t h e m e a l t o d i r e c t f l a m e o r t o steam h e a t ,  or the  product  dryers  may be p l a c e d  i n s t a t i o n a r y steam-jacketed  e q u i p p e d w i t h a r o t a t i n g s h a f t and b l a d e s meal.  f o r s t i r r i n g the  T h i s l a t t e r t y p e may be a f o r m p e r m i t t i n g  evaporation  u n d e r vacuum.  " I n g e n e r a l , t h e c o s t o f vacuum d r y i n g i s g r e a t e r t h a t o f any o t h e r the  process.  However, i n t h o s e  cases i n which  p r e s s i n g i s e l i m i n a t e d , the vacuum-drying process  costly.  than  i s less  Furthermore, t h i s l a t t e r 1-step r e d u c t i o n process  has ,  an a d d i t i o n a l a d v a n t a g e i n t h a t i t e l i m i n a t e s t h e l a r g e l o s s o f proteins that are discarded  i n the press  l i q u o r s f r o m t h e wet  process."  A l l fishmeals  c o n t a i n a c e r t a i n amount o f w a t e r - s o l u b l e  p r o t e i n which w i l l form glue o r g l u e - l i k e  s u b s t a n c e s and h a v e  a t e n d e n c y t o cake o n t o a n y h o t s u r f a c e t o f o r m a h a r d , insulating  coating.  The f a t t y f i s h m e a l s  contain  tough  sufficient  o i l t o l u b r i c a t e the hot s u r f a c e encountered i n t h e process  of  dehydration,  and  to prevent s t i c k i n g .  g l u e y m a t e r i a l s a r e removed i n t h e cooking  and  M o r e o v e r , most o f  e x t r a c t i o n o f t h e o i l by  p r e s s i n g , so t h a t t h e r e  is little  difficulty  encountered i n d r y i n g fishmeals  c o n t a i n i n g any  amount o f o i l .  n o n - o i l y waste i s p l a c e d  However, i f raw  the  appreciable in a  r o t a r y h o t - a i r d r y e r , s u c h as t h a t m e n t i o n e d a b o v e , i t e i t h e r c a k e s on t h e  s i d e s or s c o r c h e s  hardened, which prevents  and  b u r n s , o r e l s e becomes case  s a t i s f a c t o r y dehydration.  Similar  d i f f i c u l t i e s w i t h n o n - o i l y m e a l s a r e a l s o e n c o u n t e r e d i n steam j a c k e t d r y i n g ; and 1933  until  was  c o n s e q u e n t l y t h e method g e n e r a l l y a d o p t e d  t o cook t h e w a s t e , s q u e e z e out  s o l u b l e p r o t e i n s , and A n d e r s o n and  Pottinger  s t u d i e s by t h e can be  and  8  cooked r e s i d u e .  ( 1 9 ) , however, r e p o r t e d  appreciable  difficulty  i n s t e a m - j a c k e t e d vacuum d r i e r s ,  vacuum are c o n t r o l l e d d u r i n g t h e  operation.  water-  Harrison, that  "recent  a u t h o r s ...... have d e m o n s t r a t e d t h a t raw  d r i e d without  'sticking  dry the  the  These s t u d i e s  d i v i s i o n i n the  .  from 'caking' i f steam  progress  of the  suggest the  first  problems of n o n - o i l y f i s h m e a l  namely, the r e l a t i v e m e r i t  o f wet  and  waste and  pressure drying major  manufacture,  dry processes  of  reduction." (c)  Proteins The  advantages of knowing the  supplement are i n a meal are other  obvious. important  p r o t e i n value  B o t h q u a l i t y and  of  a  quantity of  i n securing proper n u t r i t i o n .  h a n d , an e x c e s s o f p r o t e i n i s n e i t h e r e c o n o m i c a l  protein On nor  the  wise, and  b e c a u s e p r o t e i n i s t h e most c o s t l y o f f e e d  too high a l e v e l of i t i n the  r a t i o n may  elements,  have a  d e l e t e r i o u s e f f e c t on t h e a n i m a l s t o w h i c h i t i s b e i n g There are  s e v e r a l f a c t o r s i n f l u e n c i n g the value  m e a l as a s o u r c e o f p r o t e i n .  o f some 2 3 amino acids©  Some o f t h e s e must be n a t u r a l l y p r e s e n t be  synthesized  from the  d i g e s t i v e agents of the  c e l l s of the  assimilated  body.  i s discarded  a c i d s w h i c h are not storage The  The  constituents.  be  a b s o r b e d and  part of the  d i s t r i b u t e d to  food w h i c h cannot  f e c e s ; and  f o r e n e r g y and  d i g e s t i b i l i t y o f p r o t e i n s may  discarded  be^ d e t e r m i n e d  breakdown.  This  other  i s d e t e r m i n e d by e s t i m a t i n g t h e  a knowledge of the  information  i s of s t i l l  greater  n e c e s s a r i l y supply  intake been  metabolic amount o f  diet,,  importance. and  product,  Even t h a t  made a v a i l a b l e t o  a l l the  a c i d s i n e x a c t l y the proper p r o p o r t i o n s  urine.  experimentally  d i g e s t i b i l i t y of the  p o r t i o n of the meal w h i c h i s d i g e s t e d t h e body does n o t  i n the  l a t t e r has  as a r e s u l t o f  w a s t e p r o t e i n on a known n o n - n i t r o g e n o u s  Besides  or  d i f f e r e n c e between n i t r o g e n  nitrogen present  be  t h e a b s o r b e d amino  the n i t r o g e n l o s s i n t h e f e c e s , a f t e r the  c o r r e c t e d f o r the  by  accessory  r e q u i r e d f o r replacement, growth  are m e t a b o l i z e d  others  down o f t h e s e m a t e r i a l s  i n the  by t a k i n g i n t o a c c o u n t t h e and  diet,  s t o m a c h , i n t e s t i n e and  o r g a n s , i n t o , a f o r m w h i c h can the  i n the  o r i g i n a l chemical  D i g e s t i o n i n v o l v e s the b r e a k i n g the  of a  I t must be remembered t h a t  p r o t e i n s are complex a g g r e g a t i o n s  can  fed.  various  f o r optimum  amino effects.  -  I t i s apparent,  9  -  then, that i t i s of v i t a l  importance  t o know  j u s t what amount o f t h e d i g e s t e d m a t e r i a l c a n be u t i l i z e d i n anabolism  and t h e l i f e  processes.  The e f f i c i e n c y w i t h w h i c h  t h e d i g e s t e d p r o t e i n s u p p l i e s t h e amino a c i d s r e q u i r e d f o r t h e c o n s t r u c t i o n o f body t i s s u e s g i v e s a measure o f t h e " b i o l o g i c a l value" of the m a t e r i a l . • T h e r e a r e s e v e r a l methods i n use f o r t h e d e t e r m i n a t i o n o f protein quality.  I n most g e n e r a l use i s t h e method o f  m e a s u r i n g t h e amount o f p r o t e i n u t i l i z e d by t h e a n i m a l i n metabolism,  by d e t e r m i n i n g t h e d i f f e r e n c e b e t w e e n t h e amount  of n i t r o g e n absorbed  and t h e n i t r o g e n d i s c a r d e d i n t h e u r i n e ,  a f t e r t h e l a t t e r has been c o r r e c t e d f o r endogenous u r i n a r y nitrogen.  T h i s i s e s t i m a t e d by d e t e r m i n i n g t h e amount o f  n i t r o g e n d i s c a r d e d on a n o n - n i t r o g e n o u s Means o f d e t e r m i n i n g  diet.  protein quality include Mitchell's  r a t b i o l o g i c a l v a l u e method  (32) ; St-*. J o h n and  b i o l o g i c a l v a l u e method, f o r a v i a n n u t r i t i o n  co-workers'  (40) ; t h e  n i t r o g e n - b a l a n c e method o f W i l g u s , N o r r i s and H e u s e r g r o w t h method o f d e t e r m i n a t i o n d e v e l o p e d Wilder  by R e c o r d ,  ( 3 7 ) ; t h e s l a u g h t e r method o f A c k e r s o n ,  Mussehl ( l ) ;  (44); the B e t h k e and  B l i s h and  t h e " g r o s s - p r o t e i n v a l u e " method i n use a t  Washington S t a t e C o l l e g e (22,38);  and more r e c e n t l y , a  c h e m i c a l method, d e v i s e d by A l m q u i s t gives a " p r o t e i n q u a l i t y index"  (2) .  and c o - w o r k e r s ,  which  -  Since these  10  -  s e r i e s were i n t e n d e d t o d e m o n s t r a t e  the  r e l a t i v e r i b o f l a v i n p o t e n c i e s of the supplements, a l l e x p e r i m e n t a l r a t i o n s had t o be b a l a n c e d  t o c o n t a i n t h e same  amount o f p r o t e i n , i • e . , 2 0 $ ( 3 0 ) .  c o n t r o l r a t i o n i n each  case  contained  were b a l a n c e d  20f°  The  s o y a b e a n m e a l ; and  the experimental r a t i o n s  by r e p l a c i n g a c e r t a i n p e r c e n t a g e  of the  r a t i o n c o n c e n t r a t e w i t h an e q u i v a l e n t amount o f t h e  control  test  material* The  p r o t e i n content  t a b l e s of c o m p o s i t i o n t i o n method, and  of the supplements i s r e p o r t e d i n the  as c a l c u l a t e d by t h e n i t r o g e n  converted to terms of "crude  determina-  p r o t e i n " by  f a c t o r 6 . 2 5 , s i n c e most o f t h e p r o t e i n s i s o l a t e d f r o m  the  animal  t i s s u e c o n t a i n s about l 6 / » n i t r o g e n ( 2 j > ) • Throughout the t r i a l s ,  s o y a b e a n m e a l was  reference source of p r o t e i n , by a number o f w o r k e r s had  since experimental  ( 2 l ) confirmed  m e r c i a l meals p r o c e s s e d  Reports  soya-  was  capable  by H a y w a r d , S t e e n b o c k  t h e f a c t t h a t , whereas t h e com-  at low temperatures  s i m i l a r i n v a l u e t o t h a t o f t h e raw  contained protein  bean, those  e i t h e r the E x p e l l e r or the H y d r a u l i c process temperatures  of  o r by a u t o c l a v i n g ,  good b i o l o g i c a l v a l u e and  of m a i n t a i n i n g normal growth. and B o h s t e d t  heat  the  data reported  indicated' that treatment  beans , e i t h e r by h e a t i n g w i t h m o i s t p r o d u c e d a m e a l t h a t had  u s e d as  prepared  by  at high  contained p r o t e i n of twice the n u t r i t i v e value  the low temperature  meal.  I t w i l l be n o t i c e d t h a t t h i s  of  is in  d i r e c t c o n t r a s t w i t h t h e f i s h m e a l s , where t h e h i g h p r o c e s s i n g  - 11 t e m p e r a t u r e was s e r i o u s l y d e t r i m e n t a l t o t h e n u t r i t i v e Hayward and H a f n e r ( 2 0 ) have p u b l i s h e d  value*  a most c o m p r e h e n s i v e  r e p o r t on t h e f i n d i n g s l e a d i n g up t o t h e e s t a b l i s h m e n t reason f o r increased n u t r i t i v e value m e a l above t h a t o f t h e raw b e a n . and Womack  o f soyabean p r o c e s s e d  (See a l s o p a p e r s by Rose  Heiman, C a r v e r and Cook  (39,46).)  chicks f e d the higher  of the  (22)  found t h a t  l e v e l s o f p r o t e i n f r o m c a s e i n and f r o m  soyabean o i l m e a l w e i g h e d e s s e n t i a l l y t h e same a t t h e end o f t h e i r experimental  period*  this laboratory (36) using meals had confirmed  Previous  experiments conducted at  commercially  p r e p a r e d soyabean o i l  t h e s e r e p o r t s , and had shown t h a t when t h e  r e s t o f t h e r a t i o n i s c o m p l e t e i n r i b o f l a v i n and m i n e r a l s , soyabean o i l m e a l may be employed as an e f f e c t i v e s u p p l e m e n t to g r a i n i n t h e c h i c k (d)  ration.  Vitamin G As e a r l y a s 1 9 3 3 i t had been r e c o g n i z e d  R i n g r o s e and N o r r i s  by W i l g u s ,  ( 4 3 ) that the n u t r i t i v e value  o f t h e more  common s u p p l e m e n t s u s e d i n p o u l t r y r a t i o n s i s c o n n e c t e d n o t only with the q u a l i t y of the proteins contained also with t h e i r vitamin G content. a method t o e v a l u a t e one  Accordingly,  i n them, b u t they  devised  these f a c t o r s separately, s i m i l a r t o the  used i n t h i s present  paper.  H e r e , t h e s e r i e s were c o n -  ducted w i t h a view t o e v a l u a t i n g t h e r i b o f l a v i n potency o f t h e t e s t m a t e r i a l s by t h e dual c r i t e r i a o f growth and t h e i n c i d e n c e of " c u r l e d - t o e " p a r a l y s i s .  The C o r n e l l i n v e s t i g a t o r s made t h e  p r o p e r a d j u s t m e n t s t o keep t h e p r o t e i n , f a t a n d bone a s h  - 12 constant,  so t h a t t h e o n l y d i f f e r e n c e i n t h e e x p e r i m e n t a l  r a t i o n was c a u s e d by v a r i a t i o n s i n t h e v i t a m i n G- c o n t e n t o f the m a t e r i a l s under s t u d y .  T h i s method was f u r t h e r  v i n d i c a t e d , b e c a u s e i n t h e c u s t o m a r y b i o l o g i c a l method o f assay (Bourquin  and Sherman, 7) o f t h i s v i t a m i n , g r o w t h i s t h e  o n l y measure o f e s t i m a t i n g t h e p o t e n c y . deduced t h a t , w i t h p r o t e i n c o n t e n t and  other  f o r gains  factors constant,  I t was t h e r e f o r e  balanced q u a n t i t a t i v e l y  v i t a m i n G was s o l e l y  i n w e i g h t above t h e c o n t r o l .  responsible  Incidence  of curled-  t o e p a r a l y s i s was o b s e r v e d a s a f u r t h e r i n d i c a t i o n o f t h e v i t a m i n potency o f t h e supplement* There a r e a t present vitamin G assays. Sherman first  conducting  The b i o l o g i c a l method o f B o u r q u i n and  ( 7 ) , e s s e n t i a l l y a r a t g r o w t h m e t h o d , was t h e one  employed.  devised  s e v e r a l methods o f  A n o t h e r b i o l o g i c a l method i n common u s e was  by i n v e s t i g a t o r s a t C o r n e l l  the e x p e r i m e n t a l  unit.  (34-),  O t h e r methods i n c l u d e  b i o l o g i c a l assay of l a c t o f l a v i n with  chicks  measurement o f t h e d e g r e e o f f l u o r e s c e n c e violet light  u s i n g t h e c h i c k as Jukes  ( 2 6 ) ; the  given by f l a v i n i n  (Supple© e t a l . , 4 2 ) ; p h o t o e l e e t r i o  measurement as d e v e l o p e d b y Cohen  9  ( 9 ) and E u l e r  fluorescence (14) and  a d a p t e d b y Hand ( l 8 ) ; t h e f l u o r o m e t r i c method o f Hodson and Norris  ( 2 3 ) ; and t h e m i c r o b i o l o g i c a l method o f S n e l l and  Strong  ( 4 1 ) , w h i c h m e a s u r e s t h e i n f l u e n c e o f f l a v i n on b o t h  the  c e l l growth and t h e a c i d p r o d u c t i o n  of Lactobacillus  e a s e l ( grown o n a s y n t h e t i c medium f r e e o f r i b o f l a v i n .  Emmet  and c o - w o r k e r s  ( 1 3 ) p u b l i s h e d a r e p o r t on t h e p r o g r e s s o f  r i b o f l a v i n a s s a y , and made a c o m p a r i s o n o f t h e s e f o u r m e t h o d s . C l o s e agreement was f o u n d among them.  The a v e r a g e s o f t h e  per c e n t d i f f e r e n c e s , i n c l u d i n g t h e s i g n , b e t w e e n t h e b i o l o g i c a l and e a c h o f t h e o t h e r t h r e e methods a r e  - 1 0 . 7 ,  + 5 . 3 , and - 7 . 0 by t h e m i c r o b i o l o g i c a l , t h e f l u o r o m e t r i c a n d t h e F l u o r a y methods r e s p e c t i v e l y .  I n t h e second s e r i e s o f  e x p e r i m e n t s , a f t e r t h e b a c t e r i o l o g i c a l method had p r o v e d t o be v a l u a b l e b e c a u s e o f t h e a d v a n t a g e  of the comparatively  s h o r t t i m e r e q u i r e d t o c o n d u c t an a s s a y , m i c r o b i o l o g i c a l d e t e r m i n a t i o n s were c a r r i e d o u t on t h e s a m p l e s study other supplementary e f f e c t s .  The f i r s t  i n o r d e r t© s e r i e s was  conducted t o e v a l u a t e t h e r i b o f l a v i n potency o f t h e c a r r i e r s .  A m o d i f i c a t i o n o f t h e method o f W i l g u s , N o r r i s and Heuser (34)  ( 4 4 ) and N o r r i s , W i l g u s , R i n g r o s e , Heiman a n d H e u s e r  was c a r r i e d o u t .  They compared /the a v e r a g e g a i n o v e r  c o n t r o l o f each r a t i o n w i t h t h e g a i n over c o n t r o l o f a standard r e f e r e n c e pork l i v e r , and e x p r e s s e d t h e v i t a m i n G c a r r i e r i n terms o f t h e pork l i v e r .  On d i v i d i n g t h e f o r m e r v a l u e b y t h e  per cent o f t e s t m a t e r i a l and m u l t i p l y i n g by 1 0 0 , t h e potency was  determined i n terms o f pork l i v e r  was  shown t o c o n t a i n 1 0 0 m i c r o g r a m s  • 100.  As t h i s  product  o f f l a v i n p e r gram, t h e  C o r n e l l " C h i c k U n i t " was d e m o n s t r a t e d t o be a p p r o x i m a t e l y e q u a l to a microgram  of f l a v i n .  S i n c e t h e t i m e t h e i r p a p e r was  w r i t t e n r i b o f l a v i n has been s y n t h e s i z e d , a n d t h e p o t e n c y o f a m a t e r i a l may now be o b t a i n e d d i r e c t l y i n t e r m s o f  - 14 micrograms of r i b o f l a v i n . W i l d e r , B e t h k e and R e c o r d  ( 4 3 ) , i n determining the  r e l a t i v e v a l u e o f f i s h m e a l s u n d e r d i f f e r e n t methods o f preparation, discovered that c e r t a i n of the processes  removed  "some o f t h e v i t a m i n G c o m p l e x " , and t h e y a l s o e m p h a s i z e d t h e importance  of c o n t r o l l i n g the v i t a m i n G content of the r a t i o n  " i f the correct conclusions regarding the protein values or the t o t a l n u t r i t i v e e f f e c t o f t h e product under a r e t o be r e a c h e d . "  investigation  T h i s p r i n c i p l e was f o l l o w e d i n t h e  present i n v e s t i g a t i o n s , but here a l l f a c t o r s , p r o t e i n , were c a r e f u l l y b a l a n c e d study t h e r i b o f l a v i n potency  including  and c o n t r o l l e d  of the meals.  i n order to  Day-old  Single-  Comb W h i t e L e g h o r n c h i c k s were p l a c e d on a r i b o f l a v i n d e f i c i e n t d i e t f o r a p e r i o d o f a week t o t e n d a y s t o d e p l e t e them o f t h e i r  store of r i b o f l a v i n .  A t t h e end o f t h i s  time  t h e y w e r e d i v i d e d i n t o b a l a n c e d l o t s and f e d t h e e x p e r i m e n t a l r a t i o n f o r f i v e weeks.  A t t h e end o f t h e e x p e r i m e n t a l  p e r i o d t h e average gains i n weight  at the three reference  l e v e l s o v e r t h e n e g a t i v e c o n t r o l were p l o t t e d a g a i n s t t h e u n i t s o f v i t a m i n G ( m i c r o g r a m s o f r i b o f l a v i n ) p e r 100 grams of feed.  From t h i s  curve t h e corresponding potency  m a t e r i a l s was d e t e r m i n e d c u l a t e d from t h e graph  The  from g a i n over  of test  c o n t r o l , and c a l -  i n terms o f micrograms of r i b o f l a v i n .  r i b o f l a v i n content of the basal r a t i o n , excluding  t h e p r o t e i n s u p p l e m e n t , was 6 3 m i c r o g r a m s p e r 100 grams; a n d o f t h e s o y a b e a n m e a l (20$) was a n o t h e r  50 m i c r o g r a m s , a s  - 15 determined i n the Strong  (41).  skimmilk,  s e c o n d s e r i e s by t h e method o f S n e l l  Soyabean meal c o n t a i n e d  2-3  and  N o r r i s et a l  micrograms;  c a s e i n , 3 micrograms per (34)  6  reported  that  a maximum w e i g h t a t f o u r weeks o f age,  3 0 0 u n i t s ; and  c r e a s i n g age, important  and  for chicks  they  require  In other  words,  requirement f o r t h i s vitamin with i n -  r i b o f l a v i n i s i n d i c a t e d as b e i n g much more  f o r r a p i d growth than f o r maintenance.  These s e r i e s  were c o n d u c t e d o v e r a p e r i o d o f f i v e w e e k s , s i n c e a t t i m e t h e r i b o f l a v i n r e q u i r e m e n t s were b e g i n n i n g and  that  to drop o f f ,  i n c i d e n c e o f a v i t a m i n o s i s as w e l l as d i f f e r e n c e s i n r a t e  o f g a i n were b e c o m i n g l e s s e v i d e n t •  Culton  and  B i r d of  U n i v e r s i t y of Maryland  (10)  crystalline riboflavin  added t o 1 0 0 grams o f b a s a l  approximately to prevent the  found t h a t 3 0 0 micrograms  1 7 5 m i c r o g r a m s p e r 1 0 0 grams was symptoms o f a r i b o f l a v i n o s i s .  not  I t was  the  of  containing sufficient concluded  that under c e r t a i n c o n d i t i o n s the r i b o f l a v i n requirement c h i c k s may  (e)  to  s i x w e e k s , about  a t e i g h t weeks, 2 9 0 u n i t s .  i s a decreasing  15;  gram*  i n order  about 3 2 5 u n i t s p e r 1 0 0 grams o f f e e d ; a t  there  and  2 0 ; I m p e r i a l Brand F i s h M e a l , 2 5 ; h e r r i n g meal,  meat m e a l , 7 » 5 ;  reach  -  be h i g h e r  than the  of  g e n e r a l l y acoepted f i g u r e s .  Minerals M a r i n e p r o d u c t s a r e good e x a m p l e s o f p r o t e i n m a t e r i a l s  associated with minerals. rich and  i n c a l c i u m and  F i s h m e a l s a r e g e n e r a l l y known t o  p h o s p h o r u s ( b e c a u s e o f t h e i r bone  a l s o t o have a h i g h e r  i o d i n e content  be  content),  t h a n most p r o t e i n  -  foods.  D a n i e l and  pointed  out  16  McCollum s t a t e t h a t Orr  that the  necessary to question. mineral sea  and  an a d e q u a t e q u a n t i t y o f t h e m i n e r a l s  in  sea has  s m a l l amounts o f most o f  elements d i s s o l v e d i n i t ,  foods w i l l prove v a l u a b l e  Iodine  i t i s t o be e x p e c t e d  sources of minerals  nutritional merits.  sodium a r e t o be  p r o d u c t s and  found i n a l l l i v i n g t i s s u e .  reported  the  presence of a l l the  was  t r a c e s o f aluminum, chromium, c o p p e r , l e a d , strontium  in addition  Newell  and  above e l e m e n t s .  d e t e r m i n a b l e by t h i s method, but  manganese, and  that  a n a l y s i s of marine  p r e s e n t i n r e l a t i v e l y l a r g e amounts i n f i s h m e a l • these,  the  I r o n , manganese, p o t a s s i u m  conducted a spectrographic  i s not  that be  Because the  McCollum ( 3 3 )  furthermore,  is  s m a l l amounts o f t h e s e p r o d u c t s w o u l d  supply  to t h e i r other  (35)  Husband  calcium-phosphorus r a t i o i n fishmeals  s i m i l a r t o t h a t o f cow's o r sow's m i l k ; and only comparatively  and  shown t o Besides lithium,  were f o u n d i n a l l o f t h e m e a l s ,  while  t r a c e s of f l u o r i n e , n i c k e l , s i l i c o n , s i l v e r , t i n , t i t a n i u m z i n c were p r e s e n t i n some o f t h e m e a l s * included  The minerals  e f f e c t of the i s not  v i t a m i n s , but  i t should  reported  above *  fishmeal manufacturing process  be  b o r n e i n mind t h a t  s o l u t i o n i n the water-soluble  on  e f f e c t on p r o t e i n s  to lose a c e r t a i n percentage of the minerals t i o n and  known i m p o r t a n c e t o  l e s s common m i n e r a l s  so p r o n o u n c e d as t h e  and  These i n v e s t i g a t o r s  i n t h e i r paper a d i s c u s s i o n of the  animal n u t r i t i o n of the  be  or  i t i s possible through  extrac-  protein portion.  - 17 (f)  -  E f f e c t s o f Method o f M a n u f a c t u r e As  e a r l y as 1929  Ingvaldsen  (24)  had  reported  that  the  method o f p r e p a r a t i o n g r e a t l y a f f e c t e d t h e  n u t r i t i v e value  fishmeals.  conducted a s e r i e s  W i l d e r , B e t h k e and  Record  (43)  of  o f e x p e r i m e n t s i n w h i c h t h e y made c o m p a r i s o n s between f i s h meals prepared under d i f f e r e n t e x p e r i m e n t a l l y c o n d i t i o n s , and  controlled  f o u n d t h a t t h e p r o t e i n s o f haddock m e a l s  duced by t h e v a c u u m - d r y i n g method were s u p e r i o r t o t h o s e the  flame-dried  meals.  The  The  previous  a f f e c t e d the b i o l o g i c a l value  of the  a b s o r b e d n i t r o g e n f r o m w e t - r e n d e r e d m e a l s was  e f f i c i e n t l y u t i l i z e d than that of the  H a r r i s o n , A n d e r s o n and  Pottinger  was  s t e a m - d r i e d haddook  M o r e o v e r , t h e method o f p r o c e s s i n g  drying profoundly  of  p r o t e i n of vacuum-dried meals  s l i g h t l y more d i g e s t i b l e t h a n t h a t o f t h e products.  pro-  dry-rendered  (19)  to meal*.  more products.  i n v e s t i g a t e d the  e f f e c t s o f v a r i o u s methods o f m a n u f a c t u r e upon h a d d o c k m e a l , a t y p i c a l n o n - o i l y f i s h m e a l , and d e f i n i t e l y decreased the animal could absorb. did  not  a f f e c t the  water-soluble  and  digestibility. seemed t o be  found t h a t  flame-drying  amount o f f i s h m e a l p r o t e i n t h a t  R e n d e r i n g by e i t h e r wet  o r d r y method  d i g e s t i b i l i t y , which i n d i c a t e d that i n s o l u b l e f r a c t i o n s had  F i n a l l y , the  the  proteins of  p r o t e i n s o f t h e head and  the similar back  e q u a l l y d i g e s t i b l e , w h i c h seemed t o i n d i c a t e t h a t  t e m p e r a t u r e was  the  protein absorption  only f a c t o r i n v o l v e d i n i n f l u e n c i n g the during the  process of manufacture.  -  The  18  -  b i o l o g i c a l v a l u e , h o w e v e r , was  by s e v e r a l f a c t o r s . decidedly  The  value  f o r the  f o u n d t o be  d r y - p r o c e s s meals  was  i n f e r i o r to that f o r the wet-process meals, which  i n d i c a t e s t h a t the w a t e r - s o l u b l e removed i n t h e m e a l , w h i c h was  proteins  l a t t e r p r o c e s s ) a r e o f low  (which would quality.  have as h i g h  The  a b i o l o g i c a l value.  d r y i n g t e m p e r a t u r e s were shown t o be o f t h e m e a l as w e l l as t o t h e apparent that flame-drying Vacuum-drying, had  no  had  detrimental  d i g e s t i b i l i t y , and  head  p a r t i c u l a r advantage over  G t h a n t h e w e t - p r o c e s s m e a l s ; and  from  High  t o the q u a l i t y made i t  a multiple detrimental  D r y - p r o c e s s m e a l s , h o w e v e r , p r o v e d t o be  p r o v e d t o be  be  e q u a l l y d i g e s t i b l e as t h e m e a l p r e p a r e d  t h e b a c k s , d i d not  of v i t a m i n  influenced  effect.  steam-drying. a better  a l s o the  a b e t t e r source of r i b o f l a v i n than the  source head  backbone,  which i n d i c a t e d t h a t the  water-soluble  t h e wet  head p r o t e i n s , were t h e most p o t e n t  process,  carriers.  and  the  In conclusion  they pointed  e x t r a c t i v e s removed  out  t h a t because  are g e n e r a l l y q u i t e s t a b l e under normal d r y i n g future w i l l be  (g)  proteins  conditions,  i m p r o v e m e n t s i n t h e method o f m a n u f a c t u r e o f  fishmeals  dependent upon methods o f i n c r e a s i n g t h e v i t a m i n  content of the  Uses o f  by  G  product.  Fishmeals  Because of t h e i r e x c e l l e n t p r o t e i n q u a l i t y , h i g h c o n t e n t and  vitamin potency, fishmeals  p o u l t r y and  swine f e e d i n g ,  c a t t l e , d a i r y cows and  and  may  be  are  of great  mineral  value  used i n f e e d i n g  in  beef  sheep, or t o s u b s t i t u t e f o r part of  the  -  milk i n r a i s i n g  dairy calves.  as s u p p l e m e n t s i n t h e  -  When f i s h m e a l s were f i r s t  r a t i o n s o f p o u l t r y and  were r e p o r t s o f t a i n t i n g i n v e s t i g a t o r s f o u n d no marine products.  19  livestock  o f t h e meat, m i l k and  such d i f f i c u l t i e s  I t seemed l i k e l y t h a t t h e  there  eggs;  from the  but  feeding  before  were l a r g e l y  their  great  value  f o r f e e d i n g was  1  have been e x t e n s i v e l y u s e d i n  p o u l t r y r a t i o n s as a s o u r c e o f p r o t e i n o f a h i g h More r e c e n t l y m i l k has o f v i t a m i n G-.  f e e d i n g , and  more p a r t i c u l a r l y  known t h a t c a r e f u l l y a biological  i n the  scraps  from the  contain  Since i t protein  comparable to t h a t of m i l k , i t  to estimate  vitamin e f f i c i e n c i e s  riboflavin  process of growth.  e q u a l l y good s o u r c e o f r i b o f l a v i n .  conducted i n order  of  in poultry  chick r a t i o n ,  u n d e r t a k e n t o f i n d out w h e t h e r f i s h m e a l s  as an  and  l a t t e r , as a m a t t e r  prepared fishmeals  value  quality.  as a good s o u r c e  f o u n d a l m o s t u n i v e r s a l use  i n t i m a t e l y concerned i n the  w h i c h has was  a l s o been r e c o g n i z e d  I t i s because of the  f a c t , t h a t m i l k has  was  realized,  Experiment  M i l k and m i l k b y - p r o d u c t s  being  for  r e s p o n s i b l e f o r t h e u n f o u n d e d reports®  SERIES (a) The  of  unstandardized  methods t h a t were u s e d i n t h e m a n u f a c t u r e o f f i s h m e a l s fertilizer  used  could  This  the r e l a t i v e  be  study  protein  o f v a r i o u s w h o l e - f i s h m e a l s and canning i n d u s t r y .  utilized was and residues  -  •(b)  20  -  M a t e r i a l s a n d Methods For the purpose o f these experiments  compiled  with a view t o determining  the r i b o f l a v i n content and  a r a t i o n was  comparative  values f o r  o f v a r i o u s B r i t i s h Columbia f i s h m e a l s  fishmeal by-products.  r a t i o n i s given i n Table  The c o m p o s i t i o n o f t h e b a s a l I.  Itwill  be s e e n t h a t w i t h t h e  e x c e p t i o n s o f t h e source o f p r o t e i n and t h e v i t a m i n G content a l l the rations are i d e n t i c a l . The  s o y a b e a n o i l m e a l was a c o m m e r c i a l l y  prepared  o f a r i c h brown c o l o u r and p l e a s a n t o d o u r and t a s t e .  brand The  r e p o r t e d a n a l y s i s ( T a b l e I I ) showed t h a t i t c o n t a i n e d 4 4 • i f . protein,  9 .67%  m o i s t u r e , 5»1lt  f a t , and  i n g r e d i e n t s were o f a s t a n d a r d t y p e poultrymen The  inBritish  5 . 7 5  /• a s h .  A l l other  i n common u s e b y t h e  Columbia.  fishmeals of these  s e r i e s were h i g h i n t h e i r f a t  c o n t e n t , c h i e f l y b e c a u s e t h e y were l a b o r a t o r y - p r e p a r e d f o r the purpose o f t h e t e s t .  T h i s was o f f s e t  by a n a c c o m p a n y i n g  h i g h p r o t e i n v a l u e , a n d was t a k e n i n t o a c c o u n t  i n the  b a l a n c i n g o f t h e rations®  of the products  in these  The p r o t e i n c o n t e n t  s e r i e s was e x c e e d i n g l y h i g h , v a r y i n g f r o m  case o f t h e m e a l p r e p a r e d  from salmon heads t o  whole h e r r i n g m e a l , and a v e r a g i n g also t h a t t h e ash content  65f».  i n the  Itwill  be n o t e d  I n t h e case o f t h e m e a l s  f r o m salmon r o e t h e v a l u e s were l o w e r ,  t h e r e was no bone p r e s e n t .  i n the  of the fishmeals varied consider-  a b l y , b u t was c o n s i s t e n t l y h i g h . prepared  over  78f»  50fo  because  Throughout t h e s e r i e s , w i t h t h e  - 21 - : e x c e p t i o n o f t h e " h e a t - t r e a t e d egg m e a l " , t h e m e a l s u s e d were prepared  i n vacuo i n a s t e a m - j a c k e t e d  A n a l y s i s of the  supplementary i n g r e d i e n t s appears i n  T a b l e II«  No  contents.  Manganous s u l p h a t e  process,  dryer*  f i g u r e s were a v a i l a b l e f o r t h e PgO^ was  and  added d u r i n g t h e  s u f f i c i e n t to ensure against the  CaO  mixing  incidence  of  perosis. The  synthetic r i b o f l a v i n , obtained  Company, L i m i t e d , was indicated.  In order to secure  r i b o f l a v i n was thoroughly  incorporated  mixed f i r s t  m i x e d and  from Merck  and  i n the r a t i o n at the  d i s t r i b u t i o n i n t h e mash, t h e  w i t h 5 pounds o f f e e d ,  s i f t e d w i t h 10  increasingly greater q u a n t i t i e s .  p o u n d s , and  The  this  so on  D o i l was  S u f f i c i e n t b a s a l was experiment to l a s t  i n c o r p o r a t e d w i t h the  prepared  at t h e ' b e g i n n i n g  throughout the  I n o r d e r t o keep r e a s o n a b l y  for  p r e p a r a t i o n of each  pounds o f mash r e q u i r e d about h a l f an h o u r o f m i x i n g . v i t a m i n A and  levels  100  The  basal. of  the  trial.  constant  a l l factors other  t h a n t h e v i t a m i n G- p o t e n c y o f t h e t e s t m a t e r i a l s , t h e f a t content  and  the mineral  content  of each of the t e s t r a t i o n s  were c a l c u l a t e d f r o m t h e l a b o r a t o r y a n a l y s e s , balanced  by a d d i n g t o them M a z o l a o i l and  I I I summarizes t h e per 100  composition  of the  and  then  bone m e a l .  experimental  p o u n d s , i n c l u d i n g t h e amounts o f f a t and  e q u a l i z e the r a t i o n s w i t h i n each s e r i e s .  Table  r a t ions  a s h added t o  I n each case  the  -  -  b a l a n c e d t o 2 0 $ , and wheat added t o make  p r o t e i n c o n t e n t was the  22  t o t a l up t o 1 0 0 pounds.  In t h i s s e r i e s the l e v e l of  p r o t e i n r e f e r s t o t h e p e r c e n t a g e o f soyabean  o i l meal  r e p l a c e d by an e q u i v a l e n t p e r c e n t a g e o f p r o t e i n f r o m a fishmeal source. In of  1 t h e r i b o f l a v i n was  Experiment  1 0 0 , 2 0 0 and 3 0 0 m i c r o g r a m s  supplements  first  f o u n d t o be  d i f f e r e n c e b e t w e e n t h e 2 0 0 and 3 0 0  (See T a b l e V I )  I t was  levels  p e r 1 0 0 grams as r e f e r e n c e  t o t h e c o n t r o l r a t i o n , but t h e r e was  no s t a t i s t i c a l levels.  fed at t r i a l  microgram  found p r e f e r a b l e a f t e r  the  experiment t o o b t a i n g r e a t e r spreads between t h e g a i n s  i n w e i g h t o f t h e c h i c k s on t h e r e f e r e n c e r a t i o n s , and quently different  subse-  l e v e l s were s u b s t i t u t e d f o r t h e s e c o n d  again f o r the t h i r d  assay.  E v i d e n t l y the l e v e l s  and  first  s e l e c t e d were a p p r o a c h i n g t h e r e q u i r e m e n t s o f t h e c h i c k s f o r r i b o f l a v i n , and t h e l o w e r l e v e l s o f 7 , 5 , 1 2 5 , and 1 7 5 m i c r o grams p e r 1 0 0 grams, and f i n a l l y 5 0 , 1 0 0 and 1 5 0  micrograms,  were s u b s t i t u t e d t o make d i f f e r e n c e s i n g r o w t h due t o  this  f a c t o r more o b v i o u s .  Day-old Single-Comb  White/Leghorn  c h i c k s were o b t a i n e d  f r o m an e s t a b l i s h e d b r e e d e r f r o m h i g h q u a l i t y have been f e d on s t a n d a r d h i g h - g r a d e r a t i o n s . were f e d a d e p l e t i o n d i e t  s t o c k known t o The  f o r a week o r t e n d a y s , and t h e n  g r a d e d on t h e b a s i s o f w e i g h t , and s e l e c t e d  so t h a t  p r i m a r y a v e r a g e w e i g h t s i n e a c h l o t were c o m p a r a b l e 85  t 5 grams.  chicks  the —  A f t e r being leg-banded f o r i d e n t i f i c a t i o n ,  they  -  were p l a c e d i n s e p a r a t e  23  -  compartments o f s t a n d a r d  brooders i n the P o u l t r y N u t r i t i o n Laboratory of B r i t i s h  battery-  a t the. U n i v e r s i t y  Columbia,  .The c h i c k s were w e i g h e d w e e k l y , b e g i n n i n g  when t h e y were  s e l e c t e d f o r w e i g h t , and a t t h e end o f each week t h e r e a f t e r * W e i g h t s were r e c o r d e d the  f o r each i n d i v i d u a l b i r d a c c o r d i n g t o  i d e n t i f y i n g leg-band.  Observations  t i m e , as w e l l a s s e v e r a l t i m e s  were made a t t h i s  d u r i n g t h e week, f o r a n y s i g n s  of a v i t a m i n o s i s or other a b n o r m a l i t i e s .  The  u s e d i n t h i s e x p e r i m e n t were so c o n s t r u c t e d i m p o s s i b l e t o keep a r e c o r d o f t h e f e e d  battery-brooders t h a t i t was  consumption, but i n  o r d e r t o e n c o u r a g e t h e maximum i n t a k e t h e t r o u g h s  were always  Lamoreux and Schumacher ( 2 ? ) r e p o r t a lOOf.  kept f u l l .  increase i n r i b o f l a v i n  i n t h e f e c e s when t h e y a r e h e l d a t  room t e m p e r a t u r e f o r 24 h o u r s , and a a r e h e l d f o r a week.  300/»  i n c r e a s e when  C a r e was t h e r e f o r e t a k e n t o  they  prevent  coprophagy because o f t h i s r a p i d s y n t h e s i s o f t h e v i t a m i n following  defecation.  M o r t a l i t y was n o t a f a c t o r i n e i t h e r s e r i e s o f experiments. accidental  The f e w f a t a l i t i e s causes  with  any p a r t i c u l a r  (c)  Results  i n Tables  were due t o  ( s u c h a s b y i n j u r y ) , and were n o t r e l a t e d ration.  The r e s u l t s o f t h i s given  that d i d occur  I V and V.  s e r i e s of three experiments are I n Table VI i s reported  a  - 24 statistical method.  summary o f t h e r e s u l t s o b t a i n e d f r o m t h e " t - t e s t "  The t e r m s S and N r e f e r t o t h e s i g n i f i c a n c e o r  n o n - s i g n i f i c a n c e o f t h e d i f f e r e n c e s between t h e means a s calculated  by t h i s method,  according to the formulae:  2  I  J  *1  /nj, + n s s  t  s  =  -  7  (n + x  2  2  +  n  +  2  1)(h + 2  2  1)  (S^ - x ) 2  s_ z where n^ and n £T(x - x) x  2  r degrees of freedom - sum o f s q u a r e s s mean.  EXPERIMENT 1 Since r i b o f l a v i n suggested i t s e l f that manufacture  i s water-soluble, the p o s s i b i l i t y s t i c k w a t e r , a waste p r o d u c t from t h e  o f f i s h m e a l by t h e w e t - p r o c e s s a l r e a d y  m i g h t p r o v e a good s o u r c e o f t h e v i t a m i n . effluent liquid  discussed,  Stickwater i s the  r e m a i n i n g when t h e o i l h a s been s e p a r a t e d f r o m t h e  p r e s s e d from t h e cooked  fish;  and s t i c k w a t e r m e a l i s  a p r o d u c t m a n u f a c t u r e d by e v a p o r a t i o n u n d e r vacuum by a patented process  (.'.28)  s  -  25  -  I n o r d e r t o make c o m p a r i s o n s between s t i c k w a t e r m e a l and other  f i s h products,  first  was a,meal p r e p a r e d  prepared was  two o t h e r m e a l s were i n c l u d e d .  The  f r o m t h e w h o l e f i s h ; t h e o t h e r , one  from t h e waste o f t h e e d i b l e p o r t i o n .  Herring  meal  s e l e c t e d f o r the w h o l e - f i s h meal, s i n c e i t i s e x t e n s i v e l y  u s e d i n p o u l t r y f e e d i n g i n B r i t i s h C o l u m b i a ; and salmon m e a l , a l s o i n common u s e , was c h o s e n as t h e m e a l p r e p a r e d cannery  from  trimmings,,  I n t h i s e x p e r i m e n t t h e c h i c k s f e d t h e 15% l e v e l o f s a l m o n m e a l r e a c h e d a w e i g h t o f 3 8 9 grams, w h i c h  represented  a g a i n o f 1 5 9 grams o v e r t h e c h i c k s on t h e c o n t r o l r a t i o n , and was c o m p a r a b l e w i t h the. 2 0 0 and 3 0 0 - u n i t l e v e l s . that very  s a t i s f a c t o r y g r o w t h was o b t a i n e d  The f a c t  would suggest t h a t  the  p r o t e i n from t h i s source  The  f a c t t h a t t h e r e was a s m a l l i n c i d e n c e o f a v i t a m i n o s i s  even a t t h e h i g h e r  was o f h i g h b i o l o g i c a l value©  l e v e l , however, would suggest t h a t  meal i s n o t s u f f i c i e n t l y potent f u n c t i o n i n g and p r e v e n t i o n  The  this  i n v i t a m i n G f o r normal  o f symptoms o f d e f i c i e n c y .  s t i c k w a t e r meal o f t h i s s e r i e s produced  slightly  i n f e r i o r w e i g h t s a t t h e 1 0 f. l e v e l , b u t t h e c h i c k s showed no symptoms o f c u r l e d - t o e p a r a l y s i s ; w h i c h w o u l d i n d i c a t e t h a t while i t i s comparatively not  rich  i n riboflavin, i t s protein i s  of p a r t i c u l a r l y high b i o l o g i c a l value.  This  is in  agreement w i t h W i l d e r , B e t h k e and R e c o r d ( 4 3 ) , who showed t h a t the w a t e r - s o l u b l e  c o n s t i t u e n t s o f f i s h m e a l s were o f  e x c e p t i o n a l l y low b i o l o g i c a l value, although  they d i d not  -  study s t i c k w a t e r meal as such. and N o r r i s  ( 4 5 ) reported  26  -  Likewise, Wilgus, Ringrose  that the stickwater  from  fishmeal  m a n u f a c t u r e , and t h e h e a d s , ( s e e E x p e r i m e n t 2 ) add materially to the vitamin Kraybill  G- p o t e n c y .  C u r t i s , Hauge and  ( 1 1 ) f o u n d a marked d i f f e r e n c e i n t h e v a l u e  hot-water-insoluble  and s o l u b l e f r a c t i o n s o f t a n k a g e s when  u s e d as a p r o t e i n s u p p l e m e n t t o c o r n * had  no s u p p l e m e n t a r y v a l u e  The s o l u b l e f r a c t i o n s  to the proteins of corn,  d e f i c i e n c i e s i n t r y p t o p h a n e and c y s t i n e , the  i n t h e wet r e n d e r i n g  due t o  " S t i c k " (which i s  product r e s u l t i n g from t h e c o n c e n t r a t i o n  obtained  of the  of the l i q u o r s  o f tankage products) i s almost  e n t i r e l y soluble i n b o i l i n g water.  This  s o l u b l e f r a c t i o n , when  f e d t o r a t s as a s o l e s o u r c e o f p r o t e i n , even a t a 15/» l e v e l , was n o t s u f f i c i e n t further and  corroborated  Pottinger  f o r maintenance.  These f i n d i n g s were  by t h e f i n d i n g s o f H a r r i s o n ,  (.19) d i s c u s s e d  Anderson  p r e v i o u s l y , who f o u n d t h a t  s o l u b l e p r o t e i n s were o f l o w q u a l i t y ; ''and t h a t meals ( c o n t a i n i n g t h e w a t e r - s o l u b l e  water-  dry-process  p r o t e i n s ) were a b e t t e r  source of r i b o f l a v i n than the wet-process meals. The  third  product under t e s t i n t h e f i r s t  h e r r i n g m e a l - p r o d u c e d much l e s s s i g n i f i c a n t and  a l s o a 6of. i n c i d e n c e  15/0  level,  of avitaminosis  experiment gains  i n weight  even when f e d a t t h e  EXPERIMENT 2 ' The  results of the f i r s t  into the n u t r i t i v e value  e x p e r i m e n t l e d t o an  of another marine  enquiry  by-product,  -  prepared  from salmon r o e .  efficiency any  -  I t was  d e c i d e d t©  o f t h i s m e a l and t o d e t e r m i n e  changes i n i t s n u t r i t i v e  t h e method o f p r e p a r a t i o n . to  27  c a r r y out  assays  the  w h e t h e r t h e r e were  value induced I t was  evaluate  by d i f f e r e n c e s i n  d e c i d e d a t t h e same t i m e  t o a c c u m u l a t e more p r e c i s e d a t a on  n u t r i t i v e v a l u e of meals prepared  f r o m salmon h e a d s  the  and  viscera. The  r e s u l t s of t h i s  IV and V. weight  T h e r e was  no  second t e s t statistical  o f t h e c h i c k s on t h e  are r e p o r t e d i n Tables d i f f e r e n c e between t h e  corresponding  m e a l u n d e r t h e d i f f e r e n t methods o f heat j u d g i n g f r o m t h e g r o w t h s t i m u l u s and n e i t h e r d i d t h e r e a p p e a r t o be any r i b o f l a v i n content.  T h i s i s not  l e v e l s of the treatment;  roe  and  i n c i d e n c e of avitaminosis,  difference in their  i n accordance w i t h  other  f i n d i n g s r e p o r t e d e a r l i e r i n t h i s p a p e r , where h i g h temperatures and  were shown t o be d e t r i m e n t a l t o b i o l o g i c a l  even more d e l e t e r i o u s t o r i b o f l a v i n c o n t e n t .  The  value cal-  c u l a t e d number o f u n i t s o f r i b o f l a v i n p e r gram o f f e e d  was  t h e same i n t h e  of  t h e raw and  egg.  case  The  o f t h e h e a t - t r e a t e d r o e m e a l , and  m e a l a p p e a r e d t o be  f a i r v i t a m i n G- c o n t e n t , and  to t h e t e m p e r a t u r e s  The  of h i g h n u t r i t i v e  t o be u n a f f e c t e d by  value  heating  i n v o l v e d i n i t s preparation©  c h i c k s w h i c h were f e d t h e m e a l p r e p a r e d  s a l m o n heads d i d not make v e r y r a p i d e i t h e r l e v e l f e d , and  from the  gains i n weight  at  a l s o showed s i g n s o f d e f i c i e n c y o f  -  riboflavin.  -  H a r r i s o n , A n d e r s o n and  t h a t h e a d s d i d not b a c k b o n e , but Likewise  28  Pottinger  have so h i g h a n u t r i t i v e v a l u e  t h a t t h e y ivere good s o u r c e s o f  W i l g u s , R i n g r o s e and N o r r i s  (45)  W i l d e r , B e t h k e and  Record  The  present  i n d i c a t e t h a t salmon-head m e a l was  as  the  riboflavin.  vitamin  (43)  w a s t e s f r o m t h e e d i b l e p o r t i o n were h i g h e r t h e heads o r t h e t a i l s .  reported  reported  i n c l u s i o n o f heads added m a t e r i a l l y t o t h e o f a m e a l , and  (19)  that  the  potency  showed t h a t  the  i n q u a l i t y than  investigation did  a good s o u r c e  not  of  riboflavin.  In the  case o f t h e  salmon-gut meal, e x c e p t i o n a l l y f i n e even a t t h e 7 . 5 $ l e v e l ; w e i g h t s w h i c h  w e i g h t s were o b t a i n e d ,  a v e r a g e d 1 8 2 grams above t h e  control.  i n c o n j u n c t i o n w i t h f r e e d o m f r o m any  The  rate  and  to contain  supply  of  riboflavin.  growth,  avitaminosis,  d e m o n s t r a t e d t h e m e a l t o have p r o t e i n o f h i g h value,  of  even a t t h e 7 * 5 l e v e l an  biological adequate  EXPERIMENT 3 I t n e x t seemed o f i n t e r e s t , s i n c e t h e m e a l f r o m salmon v i s c e r a had  p r o v e d t o be o f  experiment to study  the r e l a t i v e  from the v i s c e r a of v a r i o u s decided  at the  In the the  such v a l u e , t o r u n a  e f f i c i e n c y of meals prepared  species  of salmon.  same t i m e t o i n c l u d e two  preparation  third  It  was  l e v e l s of l i v e r  of salmon meal probably  mealo  two-thirds  l i v e r remains w i t h the head, s i n c e p a r t of the l i v e r  of lies  -  i n the s e c t i o n which  29  -  i s cut o f f w i t h t h e head,  Weights of the c h i c k s f e d the t h r e e v i s c e r a meals i n d i c a t e that these products are a l l e x c e p t i o n a l l y  fine  s o u r c e s o f h i g h - q u a l i t y p r o t e i n , and a l l good s o u r c e s  of  riboflavin. The  growth  s t i m u l u s a c c o r d e d by f e e d i n g l i v e r m e a l  i n d i s p u t a b l y h i g h e r t h a n t h a t o f any o t h e r p r o d u c t test.  The  l e v e l o f l i v e r m e a l was  5% l e v e l o f t h e v i s c e r a m e a l s o f t h i s  s i m i l a r meals; who  and  prevent  This high value  o b t a i n e d f o r l i v e r meal i s i n accordance e t a l . ( 6 ) , who  5%  and t h e  l e v e l c o n t a i n e d an a d e q u a t e s u p p l y o f v i t a m i n G- t o  Billings  under  comparable w i t h the  series;  e n t i r e l y any o c c u r r e n c e o f p a r a l y s i s .  was  with the findings  of  l a t e r made m i c r o b i o l o g i c a l a s s a y s  of  a l s o w i t h t h o s e o f Lunde o f Norway ( 2 9 ) ,  r e p o r t e d l i v e r and  r o e t o be e s p e c i a l l y r i c h  in ribo-  flavin,  (d)  Discussion As a r e s u l t o f t h e t h r e e e x p e r i m e n t s  been shown t h a t t h e f i s h m e a l s p r e p a r e d manufacturing h e r r i n g and  o f S e r i e s 1 i t has  i n ordinary  p r o c e s s e s o f the canning i n d u s t r y - namely, t h e  s a l m o n m e a l s - a r e n o t p a r t i c u l a r l y good  sources  o f r i b o f l a v i n , a l t h o u g h t h e y seem t o c o n t a i n p r o t e i n o f good biological value.  To be o f v a l u e as s u p p l e m e n t s  i n the  p o u l t r y r a t i o n t h e y w o u l d h a v e t o be f o r t i f i e d w i t h m a t e r i a l s r i c h i n v i t a m i n G-.  In the  case o f t h e b y - p r o d u c t s , h o w e v e r .  -  the r e s u l t s  30  -  s u g g e s t t h a t f u r t h e r c o m m e r c i a l u s e s h o u l d be  made o f t h e i r v a l u e  as v i t a m i n - r i c h p r o t e i n s u p p l e m e n t s .  The  v a l u e o f the, l i v e r m e a l , i n agreement w i t h t h e f i n d i n g s o f other  i n v e s t i g a t o r s , was p a r t i c u l a r l y h i g h  i n r i b o f l a v i n , and  t h e v i s c e r a m e a l s were a l s o c o n s i s t e n t l y r i c h The  two m e a l s p r e p a r e d  v a l u a b l e as a s o u r c e was  the lowest  i n this  factor.  f r o m salmon r o e were o n l y m o d e r a t e l y  o f v i t a m i n G, and t h e salmon head m e a l  of the series.  More e x t e n s i v e u s e c o u l d be  made o f t h e p r o d u c t s  rich  the  p r o v e o f b e n e f i t t o use them e i t h e r as  case.  I t should  i n r i b o f l a v i n than i s a t present  independent supplements o r i n c o n j u n c t i o n w i t h t h e o r d i n a r y fishmeals  such as t h e h e r r i n g o r salmon meals.  high r i b o f l a v i n content,  t h e s t i c k w a t e r meal s h o u l d  v a l u e when u s e d t o r e p l a c e a p o r t i o n o f o t h e r a high b i o l o g i c a l  Because o f i t s prove o f  supplements w i t h  value.  These s t u d i e s i n d i c a t e t h a t t h e q u a l i t y o f t h e f i s h m e a l s p r o d u c e d i n B r i t i s h C o l u m b i a c o u l d be enhanced i n t h e i r v i t a m i n G content  b y i m p r o v e m e n t s i n t h e method o f m a n u f a c t u r e  ( v a c u u m - d r y i n g vs» f l a m e - d r y i n g ) of f i s h r e s i d u e s  ( l i v e r , v i s c e r a , r o e and s t i c k w a t e r )  generally discarded "waste p r o d u c t s "  as w e l l as by t h e i n c l u s i o n  i n the f i s h i n g i n d u s t r y .  »  The s o - c a l l e d  have a v e r y h i g h n u t r i t i v e v a l u e w h i c h w o u l d  warrant s p e c i a l care being taken incorporation into  fishmeals.  i n t h e i r p r e p a r a t i o n and  TABLE I  Composition of Basal  Ration  E i s h O i l (100D, 1000A)  1.0  Salt  1«0  Limestone Bone E l o u r  ^. ..... •  l»j> .5  .............  Middlings  10.0  Bran , ,  10.0  Ground O a t s  10.0  Corn Meal  ., ..  10.0  Ground Wheat  ?6 .0  Soyabean M e a l . . . . . . . . . . . . . . . . a  20.0 100.0*  "Manganese s u l p h a t e a d d e d , a t t h e r a t e o f j£ pound p e r t o n o f feed„  TABLE I I Analysis of Ingredients  • Moisture  Eat  Protein  Ash  Salmon M e a l  8.91  8 . 7 6  60.75  5 . 8 0  Herring  4,21  7.84  7 8 . 6 7  9 . 6 ?  Meal  ?  2 2 . 1 0  67.40  H e a t - T r e a t e d Egg  18,20  6 0 , 9 0  3 . 5 6  Raw E g g  11,60  6 2 . 8 0  3.52  Head <>  20.40  5 0 . 2 0  i 5 c 0 3  20,60  6 2 . 3 0  7 . 8 7  71.25  7 « 8 9  13,13  72.24  7*50  Sockeye V i s c e r a  14*76  6 8 , 7 4  Liver  1 7 . 0 9  6 4 . 4 6  5.45. !  5 . 1 7  44.10  5 « 7 5  Stickwater  Meal  3.10  . Gut. Chum V i s c e r a Pink  9 . 7 3  Viscera  Meal  Soyabean O i l Meal  9*6?  1 2 , 1 5  ,.6.9.6  '  > '  ;.  2 ° 5  CaO  TABLE I I I C o m p o s i t i o n o f R a t i o n s p e r Hundred Pounds  ^Level Salmon M e a l Herring Meal Stickwater Meal  Heat-Treated Egg Raw Egg ;  Gut  Viscera Viscera  L i v e r Meal  E a t Added  Pounds  Grams  Grams  5.43 10.86.  259 322  41.0  5 , 0 0  7 . 5 0  1 2 , 5 0 5 , 0 0  4.20  202  1 0 8 . 0  1 5 . 0 0  8.40  221  1 3 5 . 0  5 . 0 0  1 5 , 0 0  3 . 2 7  129  540,0  1 0 . 0 0  10,0.0  6.54  63  2 9 8 . 0  7»50  0  5.47  1 2 0 . 0  2 0 5 . 2  .8.21  1 7 8 , 4  7 2 . 6  12.50  5,30  1 2 1 , 7  3 7 8 . 6  8 , 7 5  7*95  1 8 3 . 9  3 3 5 . 0  1 2 , 5 0  11.25  8 , 7 5  7 . 5 0  7.50  1 2 , 5 0  6 . 6 3  0  4 4 . 9  1 1 , 2 5  8 , 7 5  9 . 9 5  0  0  1 2 . 5 0  5.35  0  8 . 7 5  8 . 0 3  0  1 5 7 . 5 0  5 . 0 0  1 5 , 0 0  3 . 1 0  1 0 . 0 0  1 0 . 0 0  6 . 2 0  " 3 9 , 0  .50 25  5 , 0 0 1 0 . 0 0  Sockeye  Added  1 2 , 5 0  1 1 ,  Pink  Ash  7 , 5 0  7  Chum V i s c e r a  Supplement  1 5 . 0 0  11.2.5,  Head  Soyabean Meal Pounds  19 .1  1 6 8 . 7 .  140.6  1 5 * 0 0  3.05  2 6 . 3  ' 1 0 . 0 0 .  6 . 1 0  5 3 . 0  1 3 2 . 5 6 8 . 0 9 8 . 4  5 . 0 0  1 5 . 0 0  3 . 2 1  2 9 . 9  1 0 . 0 0  1 0 , 0 0  6,42  5 9 . 9  0  2 . 5 0  1-7.50  1.71  23*1'  1 2 1 . 6  5 . 0 0  15.00  •  3 . 4 3  1  4 5 * 4  4 9 * 0  "The " L e v e l " i n d i c a t e s t h e number o f pounds ( p e r 1 0 0 pounds) o f s o y a b e a n m e a l r e p l a c e d by a n e q u i v a l e n t amount o f p r o t e i n f r o m the source i n d i c a t e d .  :•  TABLE I T  Experimental Data  Level  Salmon M e a l  Herring  15.00  347.5 389.4  18 20 "  7.50 15.00  308*9 324.0  10 9  13 12  332.1  12 21  7 . 5 0  Meal  Stickwater  *Ave.Wt. o f Number o f C h i c k s C h i c k s a t . N o r m a l Showing 5 Wks. A v i t . G. (Grams)  Meal  5.00 ; IO.OO  Heat-Treated Egg  Raw E g g  355.8  7.50  346.62  11.25  382.05  7.50  356.33  11.25  Head  :  Chum V i s c e r a  •Pink  Viscera  •Sockeye  Liver  Viscera  Meal  15 20  Coefficient of Variability  G a i n i n Wt. Over N e g a t i v e Control (Grams)  U n i t s of VI tamin per Gram o f Eeed,  3  14.23  1 1 7 . 2  2  13»71  1 5 9 . 1  14.94  78.6  14  6.29  93.7  1 1 , 3  0  14.23 13.71  101.8 125.5  6  13.07  1  13.94  114.6 150.1  7  11.17  0  15.64,  6  27.1  •2.2.4 ' .3  33.6  Average  24.8 12.« 29.4  25.2  21,9  21.0  20.1  124.5 155.2  20.8  9 1 . 3  21.4  21.1  '12.89  2  11.03  138.4  14.3 15.1  14.7  370.42  15 19  6  11*25 7.50  413.73  23  0  9.58  181.7  37.4  37.4  8  16.89  39.2  14.08  40 .4 73.6  42.5  0  14.86 19 . 5 0  45.2  59.9  0  14.9 3 14,54  7.50  Gut  3 8 7 . 1 5  16 19  :  ,  3 2 3 . 3 3  ' 5.00 jio.oo,  3 0 3 . 6  13 20  5 . 0 0  275.2  15  270.4  10.00  291.9  2 2  3 0  5.00 10.00  289.9 28 2 . 4  18  2  2.50  279.8 306.5  19  2  1 3 . 8  22  0  17  5.00  16  Based on Normal  Chicks  3  . 50  Only.  3 6 . 0  4 8 . 5  61.9  -  -  .  52.4 49.8  76.50  •  40.0  3 1 . 5  58.3  58.3  9 3 ? ^ '  80,3  •• 6 8  ro %  TABLE V Average Weights o f C o n t r o l s (Based on N o r m a l C h i c k s O n l y ) • Wt. o f C h i c k s . a t 5 Weeks (Grams) Negative  Control  2 3 0 „ 3  Number o f C h i c k s a t 5 Weeks Normal Showing Avit. G 2  19  Basal + 1 0 0 r  3 2 6 . 7  20  0  Basal +  3 7 1 . 7  23  0  Basal + 3 0 0 ^  389.3  24  0  Negative  2 3 2 . 0  2  !?  200r  Control  Basal + 7 5  r  Gain- i n Wt-. Over Negative Control (Grams) 0  9 6.4 141.4 •  1 5 9 . 0  0  308.4  17  3  76.4  Basal + 1 2 5 ^  3 5 0 . 3  23  0  118.3  Basal +1 7 5 r  3 5 9 . 5  21  0  1 2 7 . 5  Negative  2 3 0 . 0  6  12  Basal + 5 0 f  236.4  13  6  604  Basal + 1 0 0 ^  2 5 8 . 6  19  3  2 8 . 6  Basal + 1 5 0 /  275.I  20  0  /:45»1  Control  0  TABLE V l ( a ) S t a t i s t i c a l Summary o f S e r i e s 1 Analysis of S i g n i f i c a n c e of Differences o  °. o H V  o o H O Pt  •P  o  cd ra cd  o  o  o  CM  KN  o  6  o  + >  +  H  rH cd 82 cd  cd  xa cd  PQ  « rH  © OJ  -  s  S  s  s  -.  4. B a s a l + 3 0 0  s  s  N  5 • Herring  s .  3.  Basal +  200r  7.5  s  aJ  60  •rl  £ •H  h  JH fH  ©  ffl •P  +5  o  •H  •H  H  -P  -P  • ©  e  s  s  rH  cd  o «  S  fl  o H  cd  CO .e  cd  O  rH  On  S  o - » ICN  M  O  ©  u  trs  '  tt).  cd ©  LTN  U  rH  «-•  *  s  2. B a s a l + 10G f"  o «  rH H  w  s  1. C o n t r o l  cd ©  s -s s  N s.  •  6.  Herring  15.0  s  s .; H  -  S -- - •  •  7 o S t i c k w a t e r Meal 5.0  s  -  8.  s  N  Stickwater Meal -  9 ., Salmon  7.5  10. Salmon  15.0  10.0  N  •1  s  s — ' • —  K  N _  s  -  K  N  -  TABLE V I ( b ) S t a t i s t i c a l Summary o f S e r i e s 1  j  A n a l y s i s o f S i g n i f i c a n c e of D i f f e r e n c e s  Lf\.  H O u  •p Pi o o « H  1, C o n t r o l 2. B a s a l +  -. . 75v.  + H  CM H 1 H + + • r-i H  cd xa cd  cd to cd  »  CM  M  IA CM . H  »  EH:  e  «>  C~  cd  <D w  N :  OS  S  s  O  i  M  .«  EH *  >  cd  w o  S  «  to  •  cd  9  CO  LA  s .  CM  H  « • /  lA  •  H W  w  PP  m  N  cd m cd  9 c—  10. Head 11,25  o 1A  V  s  S  s  , s,.  ;  •  3o B a s a l + 125 <  .'• s  4. B a s a l + 17 5 f  s  5. H.T. Egg  7.50  s  6. H.T. Egg  11.25  s  •• • • • 7. Raw Egg  7.5  N s  N  N  s  N : N  N.  • • s  s  8. Raw Egg 11.25  s  S  ?. Head  s  -  s  s  s s  7.5  10. Head 11.25  , - - •,11. Gut . 7 . 5  s  S  IA «  •P  s  •  TABLE V l ( c ) S t a t i s t i c a l Summary o f S e r i e s 1  • •  s  s  3. B a s a l + 1 0 0 ^ s  s  -  4. B a s a l + 1 5 0 / s  s  8. P i n k 10.0  O  -  Pi •H P4  7  <D  > •rt  <D  «  o  rH H  •H CM H  s  S.  s  • OX  IA U  .  !=•  N S  N  s  s  \  . s. s .  -  S.  S  M  .3 O  >,  o  IA CM  ,s  3.  H. : N  N.  .• . .• - •' N  N  10. S o c k e y e 10.0  12. L i v e r 5.0  &  IA  :N  9. S o c k e y e 5.0  11. L i v e r 2.5  "A •s  "*  s ; S  5.0  0  9  -  ,7. P i n k  CD  S, S, ;. S. s. . s. s  •  50< S  s  a  . tr-.  cd CO cd  2. B a s a l +  6 » Chum 10.0  *  o  <D AS o o CO  H  -  5•0  o  O H Ad Pi •H W . CO  1. C o n t r o l  5 o Chum  o  o « o H  O • IA  10.0  •+' H cd w cd m  o rH *f  Sockeye  Control  V o IA  Basal + lOOr  J  Analysis of Significance of Differences  s  ;  f  l  .  s  N  .-  s  s  -  -MISERIES 2 (a)  The  Experiment  The  first  s e r i e s d e m o n s t r a t e d t h a t f i s h m e a l s w h i c h were  s u p p l e m e n t e d w i t h s u f f i c i e n t v i t a m i n G- p r o d u c e d g r o w t h and f r e e d o m f r o m c u r l e d - t o e p a r a l y s i s . duced t h e q u e s t i o n o f t h e r e l a t i v e  satisfactory This  intro-  e f f i c i e n c y of v i t a m i n G  o b t a i n e d f r o m v a r i o u s s o u r c e s -- w h e t h e r t h e r e were any d i f f e r e n c e i n t h e e f f i c i e n c y o f r i b o f l a v i n as f o u n d i n f i s h m e a l s , i n d r i e d skimmilk or i n the s y n t h e t i c T h i s s e c o n d s e r i e s was  vitamin.  c o n d u c t e d i n o r d e r t o s t u d y t h e above  p r o b l e m , and a l s o t o e s t i m a t e t h e amount o f r i b o f l a v i n to  required  o b t a i n optimum g r o w t h and f r e e d o m f r o m symptoms o f  ariboflavinosis.  (b)  M a t e r i a l s and Methods The m a t e r i a l s  and t h e methods employed i n t h i s  second  s e r i e s were e s s e n t i a l l y t h e same as t h o s e u s e d i n t h e series. the  first  T h e r e w e r e , however, c e r t a i n m i n o r d i f f e r e n c e s i n  experimental procedure which w i l l  be d i s c u s s e d as t h e  point i n question arises. In  t h i s s e c o n d s e r i e s t h e amounts o f r i b o f l a v i n were  known f r o m m i c r o b i o l o g i c a l a s s a y . e x p e r i m e n t s was  purpose o f t h e  not t o determine the potency of v a r i o u s  c a r r i e r s , as i t was the  The  i n the f i r s t  s e r i e s , but r a t h e r t o s t u d y  r e s u l t s when known o p t i m a l amounts o f r i b o f l a v i n were  added t o v a r i o u s s u p p l e m e n t s , and t o compare t h e v i t a m i n e f f i c i e n c y of d i f f e r e n t materials with synthetic  riboflavin*  T a b l e I i n - e a c h case r e p r e s e n t s the a n a l y s i s o f the i n g r e d i e n t s used i n t h e r a t i o n s o f t h e t e s t ; T a b l e I I g i v e s the  composition of the basal r a t i o n ; Table I l i a r e p o r t s the  c o m p o s i t i o n o f e a c h o f t h e r a t i o n s p e r 50 pounds (enough f o r the  e x p e r i m e n t ) , and T a b l e I l l b ,  f o r ready c a l c u l a t i o n ,  composition of the r a t i o n s i n terms of per cent; Table g i v e s the r i b o f l a v i n c o n t e n t of each of the r a t i o n s  the I¥  as  c a l c u l a t e d f r o m m i c r o b i o l o g i c a l d e t e r m i n a t i o n s made f o r e a c h of  t h e i n g r e d i e n t s ; and T a b l e T r e p o r t s t h e w e e k l y w e i g h t  a v e r a g e s o f t h e c h i c k s on e a c h o f t h e r a t i o n s , t o g e t h e r w i t h the  (c)  incidence of avitaminosis.  Results The  for  r e s u l t s of t h i s  s e r i e s of experiments are reported  e a c h e x p e r i m e n t i n T a b l e V.  O b s e r v a t i o n s and  p r e t a t i o n s a r e i n c l u d e d w i t h each i n d i v i d u a l EXPERIMENT  The  trial,  I  first  e x p e r i m e n t was  undertaken to i n v e s t i g a t e the  e f f e c t s o f a d d i n g 225 m i c r o g r a m s of  inter-  o f r i b o f l a v i n p e r 100  feed to various l e v e l s of d r i e d skimmilk, Imperial  F i s h M e a l , h e r r i n g m e a l , meat m e a l and c a s e i n .  Any  grams Brand  Improve-  ment i n t h e r a t e o f g r o w t h o b t a i n e d upon t h e a d d i t i o n o f t h e s e s u p p l e m e n t s w o u l d t h e r e f o r e be d i r e c t l y a t t r i b u t a b l e t o t h e i n f l u e n c e o f t h e v i t a m i n , and n o t t o t h e n a t u r e o f t h e p r o t e i n .  - 33 Culton  and  B i r d (10)  had  -  reported  t h a t the  growth -  p r o m o t i n g p r o p e r t i e s o f d r i e d s k i m m i l k were g r e a t e r t h a n be  ascribed to i t s f l a v i n content.  The  a d d i t i o n of  could  dried  skimmilk to a f l a v i n - d e f i c i e n t d i e t r e s u l t e d i n greater  growth  r e s p o n s e p e r u n i t o f f l a v i n added t h a n d i d t h e  of  crystalline riboflavin* experimental  They a l s o f o u n d t h a t w i t h  b i r d s t h e y u s e d , 415  feed*  experimental  whether these r e p o r t e d t e c h n i q u e s and  2, s k i m m i l k was  crystalline 100  u s e d as a  m a t e r i a l s , i n order  to  grams o f reference  determine  f i n d i n g s were r e p r o d u c e d u n d e r  experimental  in  curled-toe  micrograms of f l a v i n per  Throughout S e r i e s  for other  prevent  n e i t h e r d i d 3 0 0 micrograms of  r i b o f l a v i n added t o 175  the  micrograms of r i b o f l a v i n  d r i e d s k i m m i l k o r d r i e d whey d i d not p a r a l y s i s , and  addition  p r o c e d u r e s employed at  the  this  laboratory*  I n E x p e r i m e n t 1, t h e s u p p l e m e n t ; and  c a l c u l a t i o n ' s were b a s e d on pounds o f  from t h i s , the  which would r e p l a c e p r o t e i n from the  amount o f p r o t e i n was  an e q u i v a l e n t  control*  determined  amount o f s o y a b e a n o i l m e a l  T h a t i s , h e r r i n g m e a l 6$  in this  e x p e r i m e n t means 6 pounds o f h e r r i n g m e a l p l u s 10.46 soyabean o i l meal per determined from the 20  100  pounds o f t o t a l r a t i o n .  f a c t that the  pounds o f This  control r a t i o n , containing  pounds o f s o y a b e a n m e a l , s u p p l i e d 8.73$ p r o t e i n .  h e r r i n g m e a l (69$ s o y b e a n m e a l was make t h e  protein)  Since  s u p p l i e d 4.161$, an amount  required to supply  t o t a l o f 8.73$ —  is  the  or i n o t h e r  6$  of  r e m a i n i n g 4.569$, t o w o r d s , 10.46  pounds  of  -  34  -  soyabean o i l meal. The then,  r e s u l t s o f t h e u n f o r t i f i e d r a t i o n s a r e due  to the  growth-stimulating  At a l e v e l of 2 $ , m i l k  p r o p e r t i e s of the  ( d r i e d s k i m m i l k ) was  prevent the occurrence of c u r l e d - t o e the higher  l e v e l s o f 4 $ and  even a t  At t h e ' l a s t l e v e l , The  entirely however,  a d d i t i o n of  micrograms of r i b o f l a v i n to the m i l k supplements d i d  p r o d u c e any  s i g n i f i c a n t g a i n over the u n f o r t i f i e d l e v e l ,  would i n d i c a t e t h a t the m i l k i n i t s e l f  is sufficiently  s u p p l i e d w i t h v i t a m i n G t o s u p p o r t good  In the  case of the  l o w e r l e v e l s o f 2 $ and to prevent the The  s u f f i c i e n t ' to  produce c h i c k s  t h e g r o w t h r e s u l t s were c o n s i d e r a b l y b e t t e r . 225  supplement.  p a r a l y s i s ; and  6 $ d i d not  f r e e from s i g n s of a v i t a m i n o s i s .  not  entirely,  well  growth.  I m p e r i a l Brand F i s h Meal l i k e w i s e , the 4%  d i d not  supply  sufficient  p r o d u c e d e x c e l l e n t g r o w t h , but  c a s e o f t h e m i l k , t h e w e i g h t s were not by t h e a d d i t i o n o f  use  which  riboflavin  c h a r a c t e r i s t i c symptoms o f v i t a m i n G d e f i c i e n c y .  6 $ l e v e l again  The  not  as i n t h e  significantly  improved  riboflavin.  o f a c o m b i n a t i o n o f 2/= I m p e r i a l B r a n d F i s h M e a l  w i t h 2% m i l k p r o d u c e d v e r y good g r o w t h r e s u l t s , w h i c h w e r e i n f l u e n c e d by a d d i t i o n a l r i b o f l a v i n .  This  confirmed  i n d i c a t i o n t h a t b o t h t h e s e p r o d u c t s were good s o u r c e s vitamin  not  the of  G.  I t w i l l be n o t i c e d t h a t where r i b o f l a v i n was  present  in  -  suboptimal  q u a n t i t i e s , the  55  -  symptoms o f d e f i c i e n c y a p p a r e n t i n  t h e t h i r d week c l e a r e d up by t h e f i f t h week, due decreasing  the  requirement f o r growth.  A t t h e 31° l e v e l , h e r r i n g m e a l was but  to  t h a t t h i s e f f e c t was  due  definitely  t o i t s l o w v i t a m i n G- c o n t e n t  not t o i n f e r i o r p r o t e i n q u a l i t y was  An  a d d i t i o n of 6f«  b r o u g h t about an improvement i n t h e r a t e  g a i n , w h i c h was  and  s t r i k i n g l y shown upon t h e  a d d i t i o n of 2 2 5 micrograms of r i b o f l a v i n . of the product  inferior,  of  l i k e w i s e much s u p e r i o r when t h e v i t a m i n  was  added.  The  same s i t u a t i o n e x i s t e d i n t h e  case o f t h e meat m e a l ,  where b o t h l e v e l s were g r e a t l y i m p r o v e d by t h e riboflavin.  The  m e a l a r e t o be Carver  and  poorer  high values  v a l u e s o f u n f o r t i f i e d l e v e l s o f meat  expected from r e s u l t s obtained  Cook ( 3 8 ) .  salmon a t 8 6 ,  T h e r e was  dried skimmilk  and  the  had  f i s h meals.  The  average "gross  uniformly  value"  of  at 9 8 ; s a r d i n e meals at 9 5 ;  no  significant  buttermilk.  and  d i f f e r e n c e between  the  Both contained  p r o t e i n of  high  g r o s s v a l u e s w h i c h were l o w e r  than those  of  Meat m e a l s were u n i f o r m l y p o o r e r  p r o t e i n , averaging value"  Robertson,  1 0 1 as compared w i t h c a s e i n , 1 0 0 ;  p i l c h a r d meals were v a l u e d  q u a l i t y , but  by  These i n v e s t i g a t o r s r e p o r t e d  from f i s h m e a l s .  h e r r i n g f i s h m e a l s was  a d d i t i o n of  only 5 5 i n gross  i s "a r e l a t i v e n u m e r i c a l  response of c h i c k s , obtained  value.  sources  This term,  e x p r e s s i o n of the  "gross  growth  w i t h p r o t e i n s u p p l e m e n t s when  added t o a d i e t b e l i e v e d c o m p l e t e i n a l l r e s p e c t s  of  except  -  36  -  q u a l i t y and q u a n t i t y o f p r o t e i n . "  The n e t g a i n p e r u n i t o f  s u p p l e m e n t a r y p r o t e i n was compared w i t h t h e n e t g a i n p e r u n i t of supplementary p r o t e i n from c a s e i n , a r b i t r a r i l y the value The  ascribed  o f 100. values  obtained  with casein i n t h i s  s e r i e s were  c o n s i s t e n t l y l o w , even when s u p p l e m e n t e d w i t h r i b o f l a v i n . comparison of the weights obtained  A  when 2 0 $ s o y a b e a n m e a l  ( c o n t a i n i n g 2 - 3 m i c r o g r a m s p e r gram, f r o m b a c t e r i o l o g i c a l determination)  was f e d , w i t h t h o s e when 2 0 $ c a s e i n  (3  micro-  grams) was g i v e n , shows t h a t t h e s o y a b e a n m e a l p r o d u c e d c h i c k s w e i g h i n g 3 6 6 , 3 8 1 and 4 0 3 grams when 7 5 , 1 2 5 and 1 7 5 m i c r o g r a m s r e s p e c t i v e l y were a d d e d , w h i l e t h e c a s e i n significantly lighter  produced  c h i c k s w e i g h i n g o n l y 3 3 0 , 3 5 7 and 3 3 9  grams a t t h e same l e v e l s . C a s e i n was f o r m e r l y  employed as a s t a n d a r d  e x p e r i m e n t s when i t was d e s i r e d t o e v a l u a t e  i n most  the q u a l i t y of a  p r o t e i n , because i n s p i t e o f a low c y s t i n e content  i t was  b e l i e v e d t o be one o f t h e most c o m p l e t e p r o t e i n s , . a n d obtained  i n a purified  form.  showed t h a t c a s e i n o b t a i n e d  However, B r a n i o n from v a r i o u s  q u a l i t y and r i b o f l a v i n c o n t e n t . i n v e s t i g a t i o n s , Almquist t o use a s t a n d a r d  et a l  c o u l d be 0  (8)  sources v a r i e s i n  Moreover, i n recent  ( 2 ) has s u g g e s t e d t h a t i t i s b e t t e r  p i l c h a r d m e a l s u p p l e m e n t as a c r i t e r i o n f o r  g r o w t h c o m p a r i s o n s , b e c a u s e t h e amino a c i d d e f i c i e n c i e s o f casein f o r chicks i n low-protein  e x p e r i m e n t a l d i e t s may  c a s e i n t o be a v a r i a b l e , dependent upon t h e amino a c i d  cause content  TABLE I Analysis of Ingredients  Moisture Herring  Meal  Ash  8.43  6 9 . 3 5  1 1 , 7 5  4 . 9 6  4 . 1 2  ;  P  2  G  5 .  .40  Soya M e a l  10,12  5 . 0 0  43.65  5 . 5 3  0 . 8 4  0  Wheat  1 2 , 5 6  2 , 2 ?  12.46  1.73  0.41  0 , 1 5  8 , 7 5  12.18  61.45  6 . 3 2  2e77  0 , 3 3  ? « 0 5  4 8 . 6 5  2 9 . 3 4  1 1 . 7 3  1 3 . 3 3  3.52  13.85  5 . 9 5  0 . 5 1  1.58  4.44-  0 . 0 5  3 4 . 3 4  1*59  2.42  1.88  1 0 . 3 ?  0 . 3 1  7 6 , 2 0  4.62  2 , 2 0  1.95  I m p e r i a l Brand F i s h Meal ;Meat M e a l Basal :  8 , 3 0  CaO  Protein  Fat  Skim Milk • Casein  6.3? 1 1 , 8 8  TABLE I I  Composition  of Basal Ration  F i s h O i l (100D, 1000A) Salt Limestone  ....  1.0  . „..... . ............... •  1.0  ......  1.5 .5  Bone F l o u r Middlings ..,  ,  10.0  Bran .  10.0  Ground Oats  10.0  Corn Meal  . . ............»  Ground Wheat Soyabean M e a l  10.0 36.0  .,. .  20 .0 100.0  ^Manganese s u l p h a t e a d d e d , a t t h e r a t e o f i - pound p e r t o n o f f e e d . •  s  TABLE, i l l - a Composition, of Rations per F i f t y Pounds Supplement Soyabean Meal Pounds Pounds  Per Cent Supplements  1.  2. 3. 4. •5. 6. 7, 8. 9. 10. 11.  12. 15. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25.  Control Soya 20/t + 75 r R i b o f l a v i n Soya 20^ + 125 / R i b o f l a v i n Soya 20$ + 175 * R i b o f l a v i n Soya 20$ + 225 ''Riboflavin Mile' Z% M i l k 4^ M i l k 6% M i l k 4$ +• 225' R i b o f l a v i n M i l k 6% + 225 R i b o f l a v i n Imperial Brand Fish Meal /  " "  r\ U  10.00  0  10,00  0  y10.OQ  0  10,  0:  10,00  1.0 2.0 3,0  2.0 3.0 1.0 2.0  2fo 4$ 6%  3.0  " Z% + . 225 ^ R i b o f l a v i n 1.0 " 4$ + 225 r R i b o f l a v i n ,2.0 Herring 3% X o 13 Herring 6% 3,0 1.5 Herring 3$-+ 225. V. " Herring 6% + 225'"' " 3.0 2.0 Meat 4$ 4.0 Meat Q% Meat 4$ + 225 ^ R i b o f l a v i n 2.0 4.0 Meat 8$ + 225^ R i b o f l a v i n Imperial Brand F i s h Meal Z% + Milk 2$ 1.0 +• LO Imperial Brand F i s h Meal 2$'+ Milk 2$- + ..225T 1.0 •+ 1,0 Riboflavin 5.73 Casein + 7 5 f R i b o f l a v i n 5.73 Casein + 125< R i b o f l a v i n 5.73 Casein + 175V R i b o f l a v i n  00  9.21 8.43 7.65 8.43 7.65 8.59 7.19 5.77 8.59 7.19 7". 62 £3 « 23 7.62 5.23 7.77 , 5,54 7.77 : 5.54  Fat Added  Ash Added  Grams  Grams  71.41 71.41 71.41 71.41 71,41 87.58 105.56 136,29 105.56 136,29  90.21 90.21 90,21 90.21 90.21 75,59 61,20 45.97 61.20 45.97  10.00 10.00 10.00 10.00 10.00 . 9.79 9.57 9.55 9.57 9.35  0 0 0 0 0 .6868 1.3736 2.0604 1.3736 2.0604  8.73 8.75 8.73 8.73 8.73 8.0433 7,3564 6.6696 7.3564 6.6696  46.70 23.18  96,64 103.49 110.32 96.64 103.49 70.08 0 70.08 0 0 0  10.41 10.81 Ho, 23 10.41 10.81 10.88  0  10.88 11.77 10*23 10.46 10.23 10.46  1.2290 2.4580 3.6870 1.2290 2.4580 2,0805 4.1610 2.0805 4.1610 1.9460 3.8920 1.9460 3.8920  7.5010 6.272 5.043 7.5010 6.272 6.6495 4.5690 6,6495 4,5690 6,7840 4.8380 6.7840 • 4.8380  -  46,70 23.18 66,83 17.94 66.83 17.94 38.32 6.73 38,32 6.75  0  V/heat to Balance Supplement Poxinds  11.7?'' •  PER CENT. PROTEIN From Supplement From Soyabean Meal  '  7.80  64.28  32,37  10.20  2.0158  6.7142  7.80 0  64,28 288.81 288.81 • 288.81  32.37 221.19 221,19 221.19  10.20 14.27 14.27 14.27  2.0158 8.73 8.73 8.73  6.7142 0 0 0  /  26. 27. 28.  Fote: —'—  0  0  B a c t e r i o l o g i c a l assay of supplements showed the following riboflavin..potencyt Soyabean Meal 2-3 * ; M i l k 20 Imperial Brand F i s h Meal 25 r ; Herring 15 Y ; Meat 7js Y\ Casein 3 r .  TABLE-111b. C o m p o s i t i o n o f R a t i o n s p e r 1 0 0 Pounds  SUPPLEMENT Ration  Lot 1. 2 . 3 . 4 . 5 . 6. 7 . 8.  9. 10* 1 1 . 1 2 . 1 3 .  14. 1 3 . 1 6 . 1 7 .  IB. 19. 2 0 . 2 1 . 2 2 . 2 3 .  24. 2 5 . 2 6 . 2 7 . 2 8 .  PROTEIN  Control Soya 2 0 / » + 7 5 * " R i b o f l a v i n Soya 2 0 f . + 1 2 5 ^ R i b o f l a v i n Soya 2 0 / . .+ 1 7 5 ^ R i b o f l a v i n Soya 2 0 f . + 2 2 5 ^ R i b o f l a v i n " M i l k 2$ " Milk 47. ;Milk 67. Milk 4$, + 2 2 5 ^Riboflavin Milk 67. + 2 2 5 r Riboflavin I m p e r i a l B r a n d F i s h M e a l 2% I m p e r i a l Brand F i s h Meal 4 f . I m p e r i a l Brand F i s h Meal 6 7 . Imp.Br.F.M. 2% + 225* R i b o f l a v i n Imp.Br.F.M. 4 / . + 2 2 5 * " R i b o f l a v i n H e r r i n g 3"/» Herring 6 $ Herring 37° + 2 2 5 ^ R i b o f l a v i n Herring 6f. + 2 2 5 ^ Riboflavin • Meat 4% Meat 8> Meat 4 7 . + 2 2 5 * R i b o f l a v i n Meat 8 f . + 2 2 5 ^ R i b o f l a v i n Imp.Br.F.M. 2% +• M i l k 2% Imp.Br.F.M. 2 f . + M i l k 2J. + 2 2 5 ^ . Riboflavin Casein + 7 5 ^ R i b o f l a v i n , Casein + 1 2 5 * R i b o f l a v i n Casein + 1 7 5 ' ' R i b o f l a v i n  Supplement Pounds 0  20  0  20  0  20  0  20  0  C  _  -...  Soyabean M e a l Pounds  20  2 . 0  18.42  4 . 0  1 6 . 8 6  6 . 0 4 . 0  15.30 16.86  6 . 0  1 5 . 3 0  2 . 0  1 7 . 1 8  4 . 0  1 5 . 3 8  6 . 0  11.54  2 . 0  1 7 . 1 8  4 . 0  1 5 . 3 8  3 . 0  15.24  6 . 0  1 0 . 4 6  3 . 0  15.24  6 . 0  1 0 . 4 6  4.0  15>54  8 . 0  11.08  4.0  1 5 . 5 4 1 1 . 0 8  8 . 0  2 . 0  +  2 . 0  15.60  2 . 0  +  2 . 0  15.60  11.46  0  11.46 11.46  0 0".  TABLE IY R i b o f l a v i n Content of Rations i n f per 100 gm»  Lot  Ration  Basal  1. Control Soya 20% * 75rRiboflavin 3. Soya ZQ% + 125 V R i b o f l a v i n 4* Soya 20% + 175 v R i b o f l a v i n 5. Soya 20$ + 225y R i b o f l a v i n •6. 13.1k 2$ 7. Milk' 4$ .. 8, 9. MilkMfo + 2251- R i b o f l a v i n 10. M i l k 6$ + 225{r R i b o f l a v i n XX » Imperial Brand f i s h Meal 2% 12. .Imperial Brand F i s h Meal 4^ 13. Imperial Brand Fish Heal 6% 14. Irap.Br.F.M. 2% + 225y Riboflavin 15. Imp.Br.F.M. A% + 225r Riboflavin 16. Herring 'Sfo 17. Herring 6% ' 18. Herring 3% + 225jT R i b o f l a v i n 19. Herring 6^ + 225r R i b o f l a v i n 20. Meat 4$ 21. Meat 8$ 22. Meat 4$ + 2253- R i b o f l a v i n : 23. Meat %$ + 225 V Riboflavin. 24. Imp.Br.E.H;- 2^ + M i l k 2$ 25. Imp.Br.F.M. 2$ + M i l k 2% + 225V Riboflavin 26. Casein + 7 5 r R i b o f l a v i n 27. Casein + 125r R i b o f l a v i n 28. Casein + 175cr R i b o f l a v i n  PROTEIN Supplement Soyabean Meal  Crystalline Riboflavin  63 65. 63 63 63 63 63 65 63 63 63 63 63 65 63 63 63 63 63 63 63 63 63. 63  0 0 0 0 0 40 80 120 80 120 50 100 150 50 100 45 90 45 90 30 * 60 30 60 50 + 40  50 50 50 50 50 46 42 38 42 38 43 35 29 43 36 38 26 38 26 39 28 39 28 39  0 75 125 175 225 0 0 0 225 225 0 0 0 225 225 0 0 225 225 0 0 225 .225  65 63 65 63  50 +.40. 34 34 54  39  225 75 125 175  o  Total  115 188 238 288 338 149 185 221 410 446 156 3J99 242 . 381 424 146 179 371 404 152 151 557 376 192 417 172 222 272  . TABLE V 'Summary of Weights-of.-Chicles in Vitamin C-.Ejp'eriment  PER CEHT PROTEIH FHOE SUPPLEMSJTS  1. 2. 3. 4. 5. 6i 7. 8. 9. -10. 11. 12. 13. 14. 15. 16. 17. 16. 13. 20. 21. -22. 23. 24. 25. 26. 27. 28.  Control Soya 20$ + ^ R i b o f l a v i n Soya 20$ + 125 J-Riboflavin Soya 20$ + 175 V Riboflavin Soya 20$ + 225'J'Riboflavin Milk 2$ Milk 4$ Milk 6$ Milk 4$ + 225 ^Riboflavin Milk 225 r Riboflavin Imperial Brand Fish Heal Z% Imperial Brand Fish Meal 4$ Imperial Brand Fish Meal 6$ Imperial Brand Pish Heal 2$ + 225VRiboflavin Imperial Brand Pish Meal 4$ •+ 225r Riboflavin Herring 3$ Herring 6$ Herring 3$ + 225» Riboflavin Herring S% + 225!r Riboflavin Heat 4jJ Meat 8$ Meat 4$ + 225 ^Riboflavin -Meat B% * 225 ITRibof lavin Imperial Brand Pish Heal zi .+ Milk 2% Imperial Brand Pish Heal 2$+Milk 2$+ ZZ5* Rioof lavin Casein 20$"+ 75'* Riboflavin -Casein 20$ + 125r Riboflavin Casein 20$ + 175 j-Riboflavin  ff/o*  FIRST WEEK SBOOED TfflEff Normal Chicks Hormal Chicks! Showing;Avlt. G gt.jgms.) Ho. m. go. - -m.. . ito 80.6 64.8 85.3 81.2 81.6 82.6 81.6 85.5 82.0 86.0 83.5 84.2 84.2 82.3' 83.5 88.9 82.4 83.6 81.9 84.8 82.7 86.7 87.4 79.9 81.1 32.5 83.8 84.7  22 21 20 21 21 22 20 21 22 22 21 20 22 20 21 18 21 19 15 19 2i 20 22 .21 22 21 18 21  123.2  28 20 124.8 20 .124.9 21. 127.8 21 141.7 20 147^8 124.3 ' 19 20 136.9 22 126.8 22 129.0 -18 112.9 15 126.9 22 131.8 20 128.8 21 134.7 14 118; 7 13 121.5 19 132.2 15 132.7 18 119.4 19 1-14.3 20 151.6 22 136.1 18 114.4 22 122.7 21' 12U2 18 127.4 21 130.7  122.7 116.0-.  121.0 142i0130.0 131.4 132.4  122.6 122.0:  123.0 111.0  4  1. 0 •0' 0 2 1 1 0 0 ". 3 5 .'•0 0 ' 0 4  8 ;00 1 2 0 0 3 0 . 0 0 0  THIRD-SEEK; Hormal -Chicks;! Showing Avit. S fft. Ho. Wt. Ho. 178.8 13 166.0 20 191.4 20 192.6 21 223.8 - 21 186.2 16 188.3 19 215.5 20 192.0 22 198.8 ' 22 • 205.9 15 203.0 14 212.7 21 200.0 20 209.9 21 179.7 14 183.7 13 212.7 19 216.1 15 168.2 17 172.3 15 1-99.7 20 . 203.4 22 •190.0. 16 203.6 22 176.9: 21 189.7 18 183.8 21  172i4163.0  187;9 207i0 212.0 196.3 196.3 196.0 164.2 169.2  154.5 162,0  9 1 0 0 0 6 " 1 1 0 0 6 6 1 0 - 0 1  -4  '8. •0 ,0 '2 6 0 0 4  0 '0 0 0  FOURTH V/EEK " Hormal .Chicks Showing-Avit. Sever Shocring Avit. . V»t.. . -Ho. V/t. Ho. W. Bo.  '245.9 5 269.0 21 278.1 20 291.7 ' 21 •287;4 20 270.5 • 20 276.5. 20 314.9 21 274.8 20 314.9 22 289.9 21 297 i'8 : 17 318,5 22 302.1 19 "316.1 21 239.3 239.1' 7 264.1 e 242.4 305 i 619 324.8' 15 16,0 .236.7 14 2 248.3' 14 219:0 291.4 '20 299.4 22 285.6 20 301.8 •22 243.4 21 260.1 :18 253.9 21  16 0 0 0 0 2 0 0 0 • 0 0 3 0 0 - 0 11 13 0 0 5 6 0 0 1 0 0 0 0  ~ftie to accident, no valiies  239.1 269.0  278a 291.7 271.7 ' 274.5 . 314.4 274.8 314.9 291.S 293.4 314.5 302.1 316.-1 247.0 26S.8 305.6 -.324.8 236.7 . 248.3 29H4 299.4•290.1 301.8 243.4 260.1 253.9  3. . 21 •: 2 0 21 0  16 •19" . 20 22 22 15 . 13 21 1C 21 6 6 19 15 14 1420 22 16 22 21 18 • ' 21 -  FIFTH YJEEK Hormal Chicks- Shooing Avit. (F" Eever Showing 'Avit. 0 Ho '.'ft. Ho. . <H; •-. ;Ko. !  365.9 381.1 404.6  21 20 21  369.3 21 366.5 20 437.4 •21 394.1 22 422.8 -. 2 2  19 390.0 19 -14 301.0 13 13 •326.3 15 14 - 281.6 16- 280.5  .385.3 452.3 22 400.4 431.3 20 308.1 -353.0 440.7 469.9 . -312.6319.5 399.7 20 . 411.8 22 418.7 21 424.4 22 330.4 21 357.3 10 339:6 21 ;  for the f i f t h ijoslc are'-available.  -0 ' 0 0 1 • 0 0  •0 0 1 0  6  0 .4  8 0 0 5 4 0 0 ' 0 0 0 0 0  :  365.9 : 381.1 -. 404.6 372.2 360.9 . .•;-• 436.0 394.1 422.8 388.9 453.3. 400.4 431.3 295.0 361.7 44Q.7 " 469.9 312.6 , 321.0 399.7 411.8. -418V9 424.4 330.4357.3 • 338.6  21 20 -. 21 - 16 . 19 20 22 22 13 . 21 19 20 6 . 6 19 15 14 14 20 22 - 16 22 21 18 •21  - 37 of the  -  r e s t of the r a t i o n ,  EXPERIMENT 2 A second experiment, s i m i l a r i n p l a n to the summer o f 1941;  undertaken i n the  but  t h i s time  s u p p l e m e n t s were c a l c u l a t e d i n t e r m s o f per the v a r i o u s  test materials,  previously.  H e r e , t h e n , 6f.  f r o m 8.64  obtained  r i b o f l a v i n c o n t e n t was reported  The  of per  i n Table  It w i l l  no  E x p e r i m e n t 1.  Fortified  the  It the  6$  of the be  unfortified  As  the  before,  with  r i b o f l a v i n by the  some  addition  of  i m p r o v i n g e f f e c t , where i t d i d i n Imperial  meat m e a l were not  Brand F i s h Meal,  other t e s t materials  not  i n the  c o n t r o l and  the  two  u n f o r t i f i e d ; and  c a n n o t be incidence 3$;  also  of the  be w i s e t o make any  by the  interpreted. of p a r a l y s i s i n  in  the  l e v e l s o f meat m e a l .  seeming d e s t r u c t i o n  means, i t w o u l d not  dried  s i g n i f i c a n t l y improved  l o t , but  because of the  the  i t w o u l d seem t h a t  seen t h a t  i s i n t e r e s t i n g t o n o t e the herring  balance of  at a l l i n accordance  a d d i t i o n o f r i b o f l a v i n , above t h e  r e s u l t s of the  protein  IT,  r i b o f l a v i n had  s k i m m i l k , and  as  d e t e r m i n e d by m i c r o b i o l o g i c a l a s s a y ,  been a d e s t r u c t i o n  u n e x p l a i n e d means.  from  cent supplement  from soyabean o i l meal.  r e s u l t s , h o w e v e r , were not  had  the  h e r r i n g meal i n d i c a t e s 6$  t h o s e t h a t xvould have been e x p e c t e d , and there  was  cent p r o t e i n  pounds o f h e r r i n g m e a l , t h e  p r o t e i n t o be made up  and  instead  first,  Hovrever,  r i b o f l a v i n by  conjectures  some  as t o  the  TABLE I Analysis of Ingredients  Eat  Moisture Herring  Meal  Protein  Ash  P 0_5  CaO  11*75  4 , 9 6  4.12  0.84  0.40  0.41  0 . 1 5  2  8*30  8 . 4 3  6 9 . 3 5  Soyabean Meal  10*12  3 . 0 0  4 3 . 6 5  5 . 5 3  Wheat  1 2 . 3 6  2 . 2 ?  12*46  1.7  1 2 . 1 8  61.45  6,32  2.77  0 . 3 3  9 . 0 5  4 8 * 6 5  29.34  1 1 . 7 3  1 3 . 3 3  3 . 5 2  1 3 . 8 5  5 . 9 5  0.51  1.58  . - 0 . 0 5  3 4 . 3 4  7 . 5 9  2.42  1.88  I m p e r i a l Brand P i s h Meal Meat M e a l Basal Skim M i l k  ,  8 . 7 3  ,  6.3?  t 1 1 , 8 8  4 .44  .  3  TABLE I I  Composition of B a s a l  Ration  F i s h O i l (100D, 1000A)  1.0  Salt  1,0  Limestone Bone F l o u r  .  1.5 ,5  ..  Middlings  10.0  B r a n . . ..  10 ,0  Ground O a t s  10.0  Corn. M e a l  10.0  Ground Wheat . „  36.0  Soyabean M e a l  .  20.0 100.0*  3E Manganese s u l p h a t e a d d e d , a t t h e r a t e o f \ pound p e r t o n o f feed©  TABLE  Ilia  Composition of Rations per F i f t y  Pounds  Supplement Pounds 1, 2. 3. 4 . 5 . 6  s  7 . 8 e  1 0 . 1 1 .  12. 13»  14. 15. 16. 1 7 . 18. 1 9 . 20. 2 1 . 22.  Control Soya 8 . • 7 - 5 % + 2 25* R i b o f l a v i n B a s a l + 37. H e r r i n g Meal Basal + 3 $ Herring Meal + 2 2 5 ^ R i b o f l a v i n B a s a l + 6 $ H e r r i n g Meal B a s a l + 6 $ H e r r i n g Meal + 2 2 5 ^ R i b o f l a v i n Basal l M Milk• Basal + iff. Milk + 2 2 5 f R i b o f l a v i n Basal + 3 > Milk Basal + 3 $ Milk + 225 Riboflavin B a s a l + 3/1 I m p e r i a l B r a n d F i s h M e a l B a s a l + 3 > I m p e r i a l Brand F i s h Meal + 2 2 5 < R i b o f l a v i n B a s a l -J" •67. I m p e r i a l B r a n d F i s h M e a l B a s a l + 6 $ I m p e r i a l Brand F i s h Meal + 2 2 5 ^ R i b o f l a v i n B a s a l + 3 7 . Meat B a s a l + 3 $ Meat + 2 2 5 R i b o f l a v i n B a s a l + 6f. Meat / B a s a l + 6f. Meat + 2 2 5 * R i b o f l a v i n B a s a l + 4-g-fo I m p e r i a l B r a n d F i s h M e a l B a s a l + 4-|fo I m p e r i a l B r a n d F i s h M e a l + - 225< Riboflavin Basal 2±% M i l k B a s a l + 2-J7I M i l k + 2 2 5 * R i b o f l a v i n  0 0  r  ..•  r  Note:  S o y a b e a n F a t Added A s h Added Meal Pounds Grams Grams 1 0 . 0 0  5 6 . 7 5  1 2 7 . 5 7 4  1 0 . 0 0  5 6 . 7 5  1 2 7 . 5 7 4  2.16 2.16  6 . 5 6  5 2 . 2 1  9 8 . 9 7 2  •6.56  5 2 . 2 1  9 8 . 9 7 2  4 . 3 2  3 . 1 3  4 7 . 6 7  4 , 3 2  47  70.143 70.143  2 . 1 8  3 . 1 3 8 . 2 8  9 5 . 3 4  9 5 . 7 9 4  2.18  8 . 2 8  9 5 . 3 4  9 5 . 7 9 4  4 . 3 6  6 . 5 6  1 3 3 . 9 4  64.014  4 , 3 6  6 . 5 6  1 3 3 . 9 4  2 . 4 4  6.56  0  2 . 4 4  6 . 5 6  0  64.014 144.77 2 144.772  3 . 1 3  0  160.716  4 . 8 8  3 . 1 3  0  160.716  3.08  6.56  3 . 0 8  6.56  8.40 8.40  6.16 6.16  3 . 1 3  0  0  3 . 1 3 4 . 8 4  0  0  4 . 8 8  3 . 6 6  -  0 0  0  1 5 2 . 5 4 4  0  1 5 2 . 5 4 4  3 . 6 6  4 . 8 4  3 . 2 7  7 . 4 2  1 0 8 . 0 5  8 0 . 1 3 1  3 . 2 7  7.42  1 0 8 . 0 5  8 0 . 1 3 1  B a c t e r i o l o g i c a l A s s a y of•Supplements showed t h e f o l l o w i n g r i b o f l a v i n potency: Soyabean M e a l , 2 - 3 / ; H e r r i n g , 1 5 / ' ; M i l k , 2 0 ^ ; I»P. ^ n d F i s h Meal,25 ; M e a t , 7 .5 * . B  .67  TABLE I H b C o m p o s i t i o n o f Rations p e r Hundred  Supplement  1. 2. 3* 4, 5 .  6. 7. 8« 9. 10. IX© 12. 1?. 14. 15.  16. 17. 18. 19. 2 0 .  21. 22.  Control S o y a b e a n M e a l 8.73f» + 2 2 5 ^ R i b o f l a v i n B a s a l + 3i H e r r i n g M e a l B a s a l + 3% H e r r i n g M e a l + 22$r R i b o f l a v i n Basal 6% H e r r i n g M e a l B a s a l + 6/. H e r r i n g M e a l + 2 2 5 ^ R i b o f l a v i n B a s a l + l-lfo M i l k B a s a l + lii M i l k +•225^ R i b o f l a v i n B a s a l + 37» Milk B a s a l + 3t Milk + 2 2 5 ^ Riboflavin B a s a l + 3% I m p e r i a l B r a n d F i s h M e a l B a s a l + 3% I m p e r i a l B r a n d F i s h M e a l + 223< R i b o f l a v i n B a s a l + 6 7 . I m p e r i a l B r a n d Eish. M e a l Basal" 6/0 I m p e r i a l B r a n d : F i s h M e a l + 225r R i b o f l a v i n B a s a l + 3% Meat B a s a l + 3% Meat + 2 2 5 R i b o f l a v i n B a s a l + 6f. Meat -• B a s a l + -6,f. Meat + ' • • 2 2 5 * ' R i b o f l a v i n B a s a l + 4-J5-/0 I m p e r i a l B r a n d F i s h M e a l Basal A-ii« I m p e r i a l B r a n d F i s h M e a l + 225< R i b o f l a v i n B a s a l + 2-ifo M i l k B a s a l + 2±f M i l k + 2 2 5 R i b o f l a v i n B  r  r  Pounds  PRO TEIN;. Supplement Pounds 0 0 4.32 4.32  8.64 8.64 4.36 4.36 8.72 8.72 4.88 4.88 9«76 9.7-6 6.16 6.16 12.32  12 . 3 2 7.32 7.32  6.54 6.54  Soyabean M e a l Pounds 10.00 10.00 13.12 13.12 6.25 6.25  16.56 16.56 13.12 13.12 13.12 13.12 6.25 6.25  13.12 13.12  6,25 6.25 9.68 9.68 14.84 14.84  p e r Hundred  Basal  Ration  Lot  .1. C o n t r o l -•2. •  3 .  4. 3 . " 6 .  i « 9. 1 0 .  11. 12.  lj. "14. -15 . •  1 6 . 1 7 . 1 8 .  19. 2 0 .  21. 2 2 ,  Soyabean M e a l 8 . 7 3 % + 2 2 5 R i b o f l a v i n Basal + 3% H e r r i n g Meal B a s a l + 3 > H e r r i n g Meal + 225*" R i b o f l a v i n Ba s a-1 6 > H e r r i n g Meal • B a s a l + b% H e r r i n g M e a l + 2 2 5 * R i b o f l a v i n Basal ItJo M i l k B a s a l + l-lfo M i l k + 2 2 5 * R i b o f l a v i n B a s a l + 37»- M i l k Basal + 3% M i l k + 2 2 5 " R i b o f l a v i n B a s a l + 3 % I m p e r i a l Brand F i s h Meal B a s a l + 3'/. I m p e r i a l B r a n d F i s h M e a l + 2 2 5 * " R i b o f l a v i n Basal + 6 > I m p e r i a l Brand F i s h Meal B a s a l + 6 f . I m p e r i a l B r a n d F i s h M e a l * -2 2 5 * R i b o f l a v i n B a s a l + 3 > Meat B a s a l + 3 % Meat + 225*" R i b o f l a v i n B a s a l + 6yo Meat - B a s a l + 6f Meat + 2 2 5 * R i b o f l a v i n B a s a l + 4i-f° I m p e r i a l B r a n d F i s h M e a l B a s a l + 4 - | % I m p e r i a l Brand F i s h Meal + 2 2 5 * R i b o f l a v i n Basal + 2j7o M i l k • Basal + 2 i % Milk + 2 2 5 Riboflavin  Supplement  Soyabean M e a l  0  50  63  0  50  65  33  63  65  33  63  130  63  130  63  87  63  87  63 63  174 174  63  1  r  PROTEIN  63 v  :  :  Grams  16 16 41 41  Crystalline Riboflavin  113  225  338  225  386  225  434  225  191 416  225  495 '  209  63  12 2  33  122  33  225  443  63  16 16  225  548  63  244 244 46 46 92 92  63 63 63  218. 323  142 .  33 33  225  367  16 16  225  171 396  63  92  12  63  92  12  63  65  18  63  65  18  '  270  63 63  -v  161 ,  33 33  Total  167 225  392  225  371  146  TABLE V Summary of Weights of Chicks August, 1941in VitaminG Experiment  I, . Soyabean -Meal -.2, oyabean Meal Meal?f8 .7 3"/•• .'+ 225 f Riboflavin -"• ••.43v.' "S Herring "•' Herring M e a l 3 % + 2 2 . 5 > ' Riboflavin "'"5, Herring Meal,6f. 6. Herring Meal bf * 225*" Riboflavin 7. Milk lH89.. ' Milk Milk 37. "+ 225^ Riboflavin 10; Bilk'37. 2 2 5 r Riboflavin .". •-,'Imperial BB rarnan dd Fish Meal 33°/= ' '• 2 2 5''-vH!bi.rl<ivlr. . ;-.II. 12; ,Imperial Fish Meal * . ' 13; Imperial' Brand' Meal: 6jl .14.- M Ini.oriul Fish V.o>\X t", ' Riboflavin eateat-3f.3/'-irun-l •15. IS-, " . M : • ' + . 225^ Riboflavin' . 17. .Meat 6f. eat Riboflavin -~ 19. M Imperial Meal 4V/. + . Riboflavin' Imperial .B Brraanndd. Fish Fishs-Heal •2i; Milk 2-H • Milk 2-Jy. -+ Riboflavin 8.75/.  •  •  •  +•  FiBh'  -  225''  <r8'.  6.7.' + ' . 2 2 5 * '  ^20.  22.  225*«'  225f  FIRST WEEK HWt orm . al Chicks •Ho. 66.7 66.2 18 66.1 19 67.0 ' 66. J 18 65.8 19 65. 19 9 66.8 5 1 65. J 67. 66.5 2 19 .66.4 , 66.4 , 66.0 18 66.5 19 66.5 19 69.6 20 66. 58.4 6 1290 58.4 19 : 57.8 20 .. 57.2 17 18  18  17  18 18 18  SECOND WEEK NWt orm . al Chicks No. 101.9 18 101.8 107.5 19 104. 7 18 109.1 117.8 19 107.8 19 105.? 17 111.6 1 112.1 19 8 112.8 J 116.6 104.4 19 101.2 20 108..7 19 IO5.I 20 100.8.." 19 , 20  18 18.  18  109.8  115.  •• 1 1 1 . 9  95.7 95.1  18 18 18  FOURTH WEEK t THIRD W E E K Chicks With Avitaminosis G Normal Chicks ! Normal .Wt. Mo. Wt. Ho. ! • Wt. Ho. 217-0 7 156.8 232.9 17 157. 8 18 3.0 19 .274.2 235.9' 1 2 9 .3 1 i 19 7 1 9 19 11 .8 164.3 19 242.2 176.3 245.1 \J 11 -L  18  19  1  •:.  ."  165.6 170.0 194.7 176.7 159.5  187.2 192.3 196.8  197.5  19  162.0 182.7  n  160.1 180.7 170,7 150.1 ' 149.5'  158.2  19  20  20  20  0 0 0  18  1  18 19 19 19 17  0  11 18  212.0 289 . 4  18 18  19 17 19 18 18 18  v«  23 5O89 . 0 264.8  j-  257  J- /  286 , 2  18 18 18  <i  29.4.1 244.4 3P3.1 •279.1 24-8 • A 235.1 255 • £ 29.2*. 27.1*. 234. 226.  L 3  18 18  230.0  19  229.7  17 18 20  19  20 17  \  284;4 4 145...2 62 4 30 59 45°i2 ,  3 0  398.4  0 0 0 0 0  FIFTH WEEK  with'Avitaminosis 8 uoriu.....u.^"Chicks aj-'^isi^-^Y : 6 », 312.7 3 2 0 0- U 0 • '  -'..  0 0 0 0 0 0 0 0 0  3 •  =^==============^  =  18  LI.:. - >  •S3  326.2  9 0  ;  369.8 334.7 366.3  19 19 19 17 • • 19 lo .-.". 1  0 0 0 0 ' 0 ' 0 0 0  ;  351.6  416.8 • 4 26;3  a  '.-  -  -  18 18  .7 l 8 4345.7 30.7 . 1915 342.8 ;16 n 371.7 420 • •401.0 333.2 i ^ 1 v  397  0  328.1  0 0  IT'  ...  20 19 20  1.1  "  310.5 '  -'.  p  302.2 ' • ,  * 0 0 0 0  • '  0  :  —  -  :  -  r e a s o n why  none o f t h e o t h e r  l e v e l s were a f f e c t e d , and  no  c o n c l u s i o n s were drawn f r o m t h i s p a r t i c u l a r e x p e r i m e n t , (Not c o n s i d e r e d i n c a l c u l a t i o n s . ) EXPERIMENT 3 A s l i g h t l y d i f f e r e n t p l a n was experiment.  the  i n the  next  B e c a u s e a c e r t a i n amount o f v a r i a t i o n was  i n E x p e r i m e n t 1, f u r t h e r work was i n order  devised  t o t e s t the  various  synthetic vitamin  continued  on t h i s  found  project  supplements to determine whether  d i f f e r e d i n a v a i l a b i l i t y from  the  combined f o r m . Once a g a i n c o n t a i n the  the  r a t i o n s were a l l c a r e f u l l y b a l a n c e d  same amount o f p r o t e i n .  experiment, 6/. fishmeal fishmeal.  i n the  to c o n t a i n ,  100  grams •  second  i n d i c a t e s 6f» p r o t e i n o b t a i n e d  Here, however, the r i b o f l a v i n  standardized u n i t s per  As  i n the  fortified  T h i s ' amount wa-s  to  c o n t e n t was level,  known f r o m  from also  368.5114.1  previous  i n v e s t i g a t i o n t o be more t h a n a d e q u a t e f o r n o r m a l f u n c t i o n i n g , so t h a t any  d i f f e r e n c e s h e r e c o u l d be  ences i n a v a i l a b i l i t y b e t w e e n t h e occurring  (combined) f o r m s .  been added i n c o n s i d e r a b l e c a l c u l a t e d and  a t t r i b u t e d to  s y n t h e t i c and  P r e v i o u s l y the excess.  same.  The  Here i t was  had  carefully  vitamin  G- v a l u e  u n f o r t i f i e d r a t i o n s would  at the  same t i m e as a f u r t h e r i n d i c a t i o n o f t h e  values  of the  supplements.  naturally  riboflavin  added i n amounts t h a t made t h e  of each r a t i o n the  differ-  serve  nutritive  -  The  4 $ d r i e d skimmilk  3?  -  p r o d u c e d , as w o u l d have been  expected from i t s r i b o f l a v i n value  of 3 2 3 micrograms, weights 2$  s l i g h t l y s u p e r i o r t o the w e i g h t s o f c h i c k s r e c e i v i n g o n l y (2l8  m i c r o g r a m s ) , and  d i d not  cause t h e  a v i t a m i n o s i s encountered i n the  small occurrence  lower l e v e l .  The  of  a d d i t i o n of  r i b o f l a v i n , however, e q u a l i z e d t h e r a t e s o f g a i n f r o m t h e  two  levels. The  same s i t u a t i o n was  f i s h m e a l and  encountered i n the  t h e meat m e a l , where t h e 4 $ l e v e l was  significantly different  from the  s l i g h t l y superior to i t . Again, was  added t o a l l t h e  2 7° l e v e l , o r was when t h e  no  m i l k p r o v e d t o he  the  either only  synthetic  not  very  riboflavin  l e v e l s t o make t h e t o t a l v i t a m i n G  i n e a c h e q u a l , t h e r e was The  case o f  value  d i f f e r e n c e between them. s u p e r i o r t o t h e f i s h m e a l and  the  meat m e a l o f t h i s e x p e r i m e n t , b e c a u s e o f i t s r i b o f l a v i n content. the  At t h e  2 f . and  v i t a m i n G content  4 $ l e v e l s f o r each o f t h e s e m a t e r i a l s , was: 2 1 8 and  Milk  F i s h m e a l ...... 145  and  Meat m e a l , 4 . . , 1 3 2 and The  176 152.  r e a s o n f o r t h i s v e r y h i g h v i t a m i n v a l u e , h o w e v e r , was,  p a r t , i t s low was  323  p r o t e i n content  n e c e s s a r y t o add  f i s h m e a l , and  8.2  11.6  ( 3 4 . 3 4 $ ) .  At the  pounds o f s k i m m i l k ,  pounds o f meat m e a l .  4$  5.7  in  levels i t pounds o f  TABLE I Analysis o f Ingredients  Moisture Fish  Meal  8*30  Protein  Ash  p o  8 . 4 3  69.35  11.75  4.96  4.12  Fat  2  5  CaO  Soya M e a l  1©>12  5 . 0 0  43.65  5.53 ,  0.84  0.40  Whe a t  12.56  2.29  12.46  1.73  0.41  0.15  61.45  6.32  2.77  0.33  11.73  13.33  .  C.F.C. S p . F i s h M e a l  8.75  Meat M e a l  6.39  9.05  48.65  29.34  l i e 88  3.52  13«85  5.95  0.51  1.58  4.44  0.05  34.34  7.59  2.42  le88  Basal Skim M i l k  12 e  18  TABLE  C o m p o s i t i o n  o f  II  B a s a l  R a t i o n  (100D, 1000A) .  F i s h  O i l  S a l t  . . . . . . .  . . . . . . . . . .  . . , .  . . . . . . . . . . . . . . . .  1.0  L i m e s t o n e B o n e  1.5  F l o u r  .5  M i d d l i n g s  . .  10.0  B r a n  10.0  G r o u n d C o r n  1.0  O a t s  M e a l  G r o u n d  10.0  . . . . . . . . . . . . . . . . . . . . . . . . . . . .  W h e a t  S o y a b e a n  10.0  .  56.0  M e a l  .'  .  20.0  100.0  ^ M a n g a n e s e o f  4  s u l p h a t e  p o u n d  p e r  a d d e d , t o n  o f  at  t h e  f e e d .  r a t e  s  TABLE I I I Composition o f Diets  PROTEIN Supplement Soyabean Meal  Supplements  Pounds  Pounds 1.  4. ',5.  FAT  ASH  Grams  Grams  Soya 8.75$  10  65.0712 85.3974  Soya 8 . 7 5 $ + 65J R i b o f l a v i n  10  65.0712 85.3974  Soya 8,73$ + 130^ R i b o f l a v i n  10  65.0712 85.3974  Soya 8.73$ + 260^ R i b o f l a v i n  10  65.0712 85.3974  M i l k 2$  WHEAT TO BALANCE SUPPLEMENT Pounds  2.912.  7.705  116.2240 42.7668)  5 a . M i l k 2$  2.912  7.705  116.2240 42.7668)  6.  M i l k 2$ + 160?" R i b o f l a v i n  2.912  7.705  116.2240 42.7668)  6a. M i l k 2$ + 1 6 0 / R i b o f l a v i n  2.912  7.705  116.2240  7.  M i l k 4$  5.824  5.415  167.5487  8..  M i l k 4$ + 6 0 ^ R i b o f l a v i n  5 0 8 24  5.415  ?.  F i s h 2$  1.442  7.705  61.7213 66.1478) )  9a.  F i s h 2$  1.442 • "  7.705  61.7213 66.1478)  9.383  )  9.383  )  42.7668) 8.76I  0  167.5487  0  10.  F i s h 2$ +  21$fRiboflavin  1.442  7.705  61.7213 66.1478)  10a.  F i s h 2$ + 215^ R i b o f l a v i n  1.442  7.705  61.7213 66.1478)  11.  F i s h 4$  2.883  5.415  58.5433 46.8982)  12.  F i s h 4$ + 1 7 0 y R i b o f l a v i n  2.883  5.415  58.5433 46.8?82)  15.  F i s h 6$  4/325  3.126  49.8492 21.2245)  14.  F i s h 6$ + 125 if R i b o f l a v i n  4.325  3.126  49.8492 21.2245)  15.  Meat 2$  2.055  7.705  32.4610  0  1 5 a . Meat 2$  2.055  7.705  32.4610  0  10.855  )  )  Riboflavin  2.055  7.705  32.4610  0  16a.  Meat  Riboflavin  2.055  7.705  32.4610  0  17.  Meat 4$  4.110  5.415  0  0  18.  Meat 4$ +  4.110  5.415  0  0  Note;  +  230cT^  200?r  Riboflavin  12.545  )  9.760  Meat 2 $ + 23Qf 2 $  11.705  )  16.  !  10.855  9.760  9.525 )  B a c t e r i o l o g i c a l a s s a y o f s u p p l e m e n t s showed t h e f o l l o w i n g r i b o f l a v i n potency: Soyabean M e a l , 2 - 3 r ; M i l k , 20 f ;F i s h , 15 r ; Meat 7 ^ r . ;  Ilia.  TABLE  Ilia.  Composition of Rations  p e r 100 Pounds  SUPPLEMENT Lot  1. 2. 3 .  4. 5. 5a. 6. 6a. 7. 8.  ?..  9a. 10. 10a. 11. '12. 13. 14. 15. 15a. 1 6 .  16a. 17. 18.  Ration  Soya Soya Soya Soya Milk Milk Milk Milk Milk. Milk Fish Fish Fish Fish Fish Fish Fish Fish Meat Meat Meat Meat Meat Meat  8.73%  8 . 7 3 % + 65 * R i b o f l a v i n 8.73% + 1 3 0 * Riboflavin 260rRiboflavin  8 . 7 3 % +  H i i i 4% 2 2  + 160 *" R i b o f l a v i n  2  +  l60*Riboflavin  6 0 * Riboflavin 4% + 2% 2% 2% + 215 R i b o f l a v i n 2% + 215 R i b o f l a v i n " ' 4% + 170 * R i b o f l a v i n A r  Y  i i 6%  6  + 125 r R i b o f l a v i n  2% 2% 2 i + 2% +  i  PROTEIN Supplement Pounds 0 0 0 0 5.824 5.824 5.824 5.824  1 1 . 648 11.648 2.884 2.884 2.884 2.884 5.766 5.766 8.650 8.650 4.110 4.110  2 3 0 r- R i b o f l a v i n 230>-Riboflavin  A  4% + 200 r  Riboflavin  4.110 4.110  8.220 8.220  Soyabean M e a l Pounds 20 20 20 20 15.410 15.410 15.410 15.410 10.830 10.830 15.410 15.410 15.410 15.410 10.830 10.830 6.252 6.252  15.410 15.410 15.410 15.410 10.830 IO.83O  TABLE IT R i b o f l a v i n Content of Rations In r per 100 gm.  Lot  1. 2. 3. 4. 5. 5a. 6. 6a. 7. 8. 9. 9a. 10. 10a. 11* 12. 13. 14. 15. 15a. 16. 16a« 17. 18.  Ration  Basal  PROTEIN Supplement Soyabean Meal..  Soya 8.73$' 0 63 Soya 8.73$ + 65 *r R i b o f l a v i n 0 63 Soya 8.73$ + 1 3 0 r R i b o f l a v i n 0 63 Soya 8.73?y + 260*" R i b o f l a v i n 0 63 M i l k tC/o 63 116.5 Ifilk 63 116.5 I f i l k d/o + 160V R i b o f l a v i n 63 116.5 116.5 M i l k 2% 4- 160V R i b o f l a v i n 63 Milk. 4% 63 235 M i l k 4$ + 60er R i b o f l a v i n 63 235 63 43 F i s h 2% 63 F i s h 2% 45 P i s h 2% + 2T5r R i b o f l a v i n . 63 45 45 F i s h 2% -f 215s- R i b o f l a v i n 63 86 F i s h 4$ 63 86 63 F i s h 4$ + 170-r R i b o f l a v i n 63 > 193 F i s h 6$ 63 193 F i s h erf O/O 63 31 Meat 2% 31 63 Meat 2% Meat 2% + 230v R i b o f l a v i n 65 31 63 31 Meat 2% + 230r R i b o f l a v i n 62 63 Meat 4$ 62 63 Meat A% + 200V R i b o f l a v i n  50 50 50 50 38.5 38.5 38.5  "  27 27 38.5 ' 58.5 38.5 38.5 27 27 15 15 38.5, 38.5 .. .58.5 58.5 27 27'  \ -  Crystalline Riboflavin 0 65 130 260 0 0 160 160 0 60 0 0 215 215 0 170 0. 125 0 0 230 230 0 200  Total  113 178 243 373 218 218 378 378 525 383 145 145 360 360 176 346 271 396 132 132 362 -,. 562 152 552  TABLE T Summary of Weights of Chicks i n Vitamin G Experiment October, 1941.  SOEKLEIEHTS  1. 2. 3. 4. 5. 5a. 6. 6a. ' 7. 0. 9. 9a. 10. 10a. 11. 12. 13. 14. IB. 15a. 16. ;l6a. 17. 18.  Ho. of : HormalChicks --Showing Avit.-G Chicks Ho. Wt. Ho. , 7ft.  75.4 21 Soya 8.73$- •< - , . 22 Soya 8.73$ + 65 K Riboflavin : ,- -77.0 76.4 .22 Soya 8.73$+ 130r Riboflavin 22 Soya- 8.73$ +2S0ir Riboflavin.! 75.3 Milk 2$ . 75.7,: 21 .77.9 - 21 Eilk 2$ , 22 SUlk -2$ + 160r Bibof lavin- V , 78.9 82.8 20 Milk ,Z% * 1600iboflavin Hilk 4$ - 21 - 73.5 Milk 4$ + eo/Riboflavin 75.6 - '21 Pish 2$ . • • 74.1 21 Pish 2$ ,22 -. -78.0 22 Pish 2$-+ 215if-Riboflavin • .' 78.1 Pish 2$ +215/ Riboflavin -21 77.4 Pish 4$ ' ' . 22 74.8 Pish 4$ + 17orRiboflavin 76.4 21 Fiah 6$ 20 • -78.3 Fish'6$.+ 125»' Riboflavin .' 76.8 20 Heat -2$ - . -. 75.0 22 Kent 2$ - 78.6. 21 -Meat 2$ + 230><t Riboflavin .-: - - '75*0 ,22 Moat 2$ + 230yRibofiavin 82.2 22 Moat 4$ .' . 79.1-, "22 Meat 4$ + 200r"Ribof lavin 76.3 22  FOURTH WEEK  THIRD -'.VEESI  • TEH DAYS. Initial wt.  143.0 163.1 -160.9 .174.6 .166.2. .155.4 176.8 174.9 174.2 169.1 146.7 139*9 .189.3 177.0 -143;7 183.0 144*6 , 169.8 146.3 157.9 '174.3 174.0 146.9 175.5  Hormal Chicks Wt.  7 • 186*8. 143.0 13 151.0 232.2 184 - - . ' .0 22 241.3 , , .:•-. 249.0 21 0 249.2 3. 18" 175.3 ' 11 159. .3-: 3 - 254.8 0 259.6 21 , • . 0 . 258.6 19 259.1 0 21 254.6 0 21 9 202.7 137.3 11 •135.5 181.8 12 10 ' :• -. • o - 283.4 22 . 0 270.3 21 144.5 16 203.0 6 274.1 210 203.1 7 - 131.4 13 262.9 19 ' 174.0 1 9 200.3 12 140,3 239.1 17 • 147.0 -4 254.7 22 0 22 0 259.3 178.6 146.5 ' 6 -16 22 0 . 259.5  Ho. 12 19 20 22 21 17 21 20 21 20 3 4 23 20 5 17 6 20 12 15 22 22 20 22 .  Hever'Showing Avit.G Wt. 191.5 233.7 241.3 249.0 245.3 254.2 259.6 258.6 259.1 254.6 202.7 181.8 283.4 270.3 209.8 274.1 209.8 261.8 195,0 239.0 254.7 259.3 178.8 259.5  FIFTH --TCEEK-. Showing Avit  - Ho. : 11 17 20 22 16 15 21 20 21 20 3 4 23 • 20 4 17 4 19 9 14 ; 22 22 16 22  Wt.  Ho  196.2 201.3 -  6 3 0 0 0 0 0 0 . 0 0 16 17 0 0 15 0 14 .. 0 9 4 -0 0 0 0  -  -  - -  189.1 192.9 -  169.5 179.6 189.2 212*0 -  -  Normal Ghicks Kales ••"-Females .'.Average wt. 232.0 253.0 299.-9 302.4 285.6 -308.5 • 335.0 • 290.0' 351.9 324*0 :349.3 301,6 ,364.8 326.6 - 366.3 -335*1 378.9 314.8 •' "368.0 352.6 275.3 .259.4 254.0 280.0 402.7 •370.0 387.1 338.0 284.0. 285.8 390.6 . -349.1 282.0 . 276.0 377.6 •347*4 241.0 272.7 275.1 . •316.9363.7 : 327.9 325.5 378.9. 259.4 • 235.9 319*0. 373.4  Hever Showing Avit. G Males Females -Average Wt. Wt. Wt.  253.0 242.5 218.4 305.5 301.4 303.0 308*5 297.1 285.6 290.0 307.6 335.0 324.0 337.1 342.3 301.6 326.6 349.3 326.6 •364.8 346.6 350.7 366.3 ' -335.1 314.8 378.9 348.4 352.6 359,5 - 368.0 266.0 264.2 i 279.5 266.7 271.5 - 254.0 370.0 392.8 402.7 338.0 371.6 - 387.1 300.0 286.0 290.0 349.1 368.6 390.6 292.7 277.8 272.0 347.4 362.5 377.6 278.5 241.0 270.2 316.9 292.3 • 281.1 327.9 337.7 363*7 325.5 378.9 347.4 235.9 259.4 245.3 319.0 348.7 373.4  237.3 304.5 297.1 307.6 332.1 326.6 346.'6 350.7 348.4 359.5 272.5 263.6 392.8 .371.6 295.0 368.6 287.5 362.5 274.3 297.8 337.7 347.4 245.3 348.7  Showing Avit. G Males Females Average Wt. Wt. Wt. 231.7  230.0  231.3  262.4 248.7  234.0 207.3  251.7 240.4  255.9  262.0  257*9  190.3  254.4  225.9  248.6 283.0  245.6  247.1 283.0  Hever Afflicted Males Females -Ho. - .Ho. 5 17 20 9 11 1-111 •10 --11 9 2 2 16 13 3 13 3 •10 1 10 6  6 12 11 14 10 10 10 -10 -10 11 2 .'4 7 • 6/ 4 8 7 10 12 7 16 IS 12 10  - Afflicted Hales. Female Ko. Ho.- . 3 2 0 ' 0 0 0 0 0 -0 0 7 12 0 : 0 8 ' 0 4 '0 5 3 0 0 0 0  - Recovered Hales .Feaale Ho. - Ho* - .1 2 0 0 3 0 0 0 0 0 . 1 1 0 0 ' 2 , 0 2 0 '0 2 0 0 0 0  (d)  B j s cuss i o n From t h e t h r e e e x p e r i m e n t s c a r r i e d out i n S e r i e s 2 i t has  been shown t h a t p r o t e i n s u p p l e m e n t s are  fortified  with  e q u a l l y as good as s u p p l e m e n t s n a t u r a l l y r i c h  riboflavin in this  factor. The  first  riboflavin  e x p e r i m e n t was  c a r r i e d out by a d d i n g s y n t h e t i c  i n e x c e s s o f t h e known r e q u i r e m e n t s , s u p p l e m e n t i n g  one member o f each p a i r o f r a t i o n s a t a g i v e n l e v e l w i t h 2 2 5 u n i t s of r i b o f l a v i n .  H e r e t h e p r o t e i n v a l u e s were g i v e n i n  terms o f per cent supplement. on t h e u n f o r t i f i e d the  Regardless of the f i n a l  results  r a t i o n s , the chicks attained approximately  same w e i g h t s upon t h e a d d i t i o n o f an e x c e s s o f t h e v i t a m i n .  S k i m m i l k i n c l u d e d i n t h e t e s t was  no more e f f i c i e n t  ini t s  growth-stimulating properties than other r a t i o n s equal i n p r o t e i n and s u p p l e m e n t e d  with the synthetic  riboflavin,  c o n t r a r y t o r e s u l t s r e p o r t e d by some o t h e r investigators©  The the  s e c o n d e x p e r i m e n t was  c a r r i e d o u t by s u p p l e m e n t i n g  r a t i o n s w i t h s y n t h e t i c r i b o f l a v i n as i n t h e p r e v i o u s one,  b u t h e r e t h e p r o t e i n v a l u e s were c a l c u l a t e d i n t e r m s o f p e r c e n t p r o t e i n , f o r e a s e o f c o m p a r i s o n on a p r o t e i n b a s i s . was  There  some u n e x p l a i n e d d e s t r u c t i o n o f r i b o f l a v i n , h o w e v e r , w h i c h  r e n d e r e d t h e r e s u l t s o f t h i s e x p e r i m e n t beyond The  t h i r d e x p e r i m e n t was  interpretation.  d e v i s e d so t h a t one r a t i o n o f  each p a i r of r a t i o n s at a g i v e n l e v e l c o n t a i n e d a d e f i n i t e amount o f r i b o f l a v i n , words,  certain  368+14.1 m i c r o g r a m s .  i n s t e a d of a d d i n g 2 2 5 micrograms  t o each  In other  level,  - 41 i r r e s p e c t i v e o f i t s o r i g i n a l v i t a m i n c o n t e n t , an i n d e p e n d e n t l y calculated  q u a n t i t y was added t o each i n d i v i d u a l r a t i o n .  The  ;  p r o t e i n was a g a i n c a l c u l a t e d i n terms o f p e r c e n t p r o t e i n a given source.  As a r e s u l t o f t h i s  from  e x p e r i m e n t i t was shown  t h a t v i t a m i n G was e q u a l l y a v a i l a b l e w h e t h e r  o b t a i n e d from  n a t u r a l sources o r from t h e s y n t h e t i c p r o d u c t .  No improvement  t h r o u g h t h e f e e d i n g o f d r i e d s k i m m i l k was n o t e d , w h i c h c o n f i r m e d t h e c o n c l u s i o n s drawn f r o m t h e r e s u l t s o f E x p e r i m e n t 1, Moreover, adequate  3 7 0 micrograms  p e r lOO grams o f f e e d was more t h a n  f o r n o r m a l g r o w t h and m a i n t e n a n c e , and t h e minimum  requirements f o r s a t i s f a c t o r y  growth were 2 2 5 m i c r o g r a m s .  O b s e r v a t i o n s and comments on t h e i n d i v i d u a l  supplements  u s e d i n t h i s s e r i e s a r e i n c l u d e d w i t h e a c h e x p e r i m e n t , and the r e s u l t s  summarized i n t h e t a b l e s .  F i g u r e 1 r e p r e s e n t s t h e " s c a t t e r d i a g r a m " o b t a i n e d by p l o t t i n g , f r o m t h e d a t a o f E x p e r i m e n t s 1 and 3, t h e w e i g h t s o f c h i c k s i n grams a g a i n s t t h e number o f u n i t s o f r i b o f l a v i n f e d . The mean o f t h e c h i c k w e i g h t s was c a l c u l a t e d , f r o m 49 i n d e p e n d ent v a r i a b l e s , as b e i n g 357.14+56.42 ( c o e f f i c i e n t o f v a r i a b i l ity  =  1 5 . 8 0 % ) ; and t h e mean o f t h e c o r r e s p o n d i n g l e v e l s o f  r i b o f l a v i n a s 267 .45+.102.23 ( c o e f f i c i e n t  of v a r i a b i l i t y -  38.22%). The c o r r e l a t i o n c o e f f i c i e n t , r , was c a l c u l a t e d f r o m t h e equation:  2SO  ,  26 O  300  o  O  •too  56o  o  ft  O  '  o X \  X •  t  X  oo N  .  XXX  X  V> M  o  « ft  °\  X  X  o X  X  o  o  o  X  "I .1  X s  o In  o o  j'o I*  :  id  2 e  o  \  o o  o  X  X  o  X o *>  X  \  \  X  x  IS  XX  1  X  Figure 1  - 42 -  r  xy =  5  (  * " S).(y^- )A <r x . <r y y  =  +0,4843  This i n d i c a t e d a h i g h l y s i g n i f i c a n t p o s i t i v e  correlation  between g a i n i n w e i g h t and m i c r o g r a m s o f r i b o f l a v i n f e d . W i t h t h i s i n f o r m a t i o n , t h e r e g r e s s i o n c o e f f i c i e n t was d e t e r mined f r o m t h e f o r m u l a :  y X  b  '  xy  £(x - x K  =  S l l L . - y) (* - £). = G.S776 E(y - y ) Z  For  each i n c r e a s e i n 1 u n i t r i b o f l a v i n t h e r e  i s a correspond-  i n g i n c r e a s e o f .2673 grams i n w e i g h t ; and c o n v e r s e l y ,  each  gram g a i n i n w e i g h t w o u l d r e q u i r e t h e a d d i t i o n o f micrograms o f r i b o f l a v i n . b y , i srepresented X  The r e l a t i o n between t h e v a r i a b l e s ,  by t h e r e g r e s s i o n l i n e i n t h e f i g u r e .  SUMMARY Two s e r i e s o f f e e d i n g e x p e r i m e n t s were c o n d u c t e d i n order t o determine t h e n u t r i t i v e value o f B r i t i s h fishmeals  Columbia  i n chick feeding.  In the f i r s t  s e r i e s , which consisted o f three  experiments i n v o l v i n g t w e n t y - t h r e e l o t s - o f twenty-two  chicks  e a c h , t h e n u t r i t i v e v a l u e o f e l e v e n f i s h m e a l s and b y - p r o d u c t s o f t h e f i s h i n g i n d u s t r y was d e t e r m i n e d .  The h e r r i n g and  -  43  -  salmon m e a l s under i n v e s t i g a t i o n were not p a r t i c u l a r l y s o u r c e s o f r i b o f l a v i n , but L i v e r m e a l was  c o n t a i n e d p r o t e i n o f high, q u a l i t y .  p a r t i c u l a r l y r i c h i n the v i t a m i n G f a c t o r ,  the t h r e e meals prepared s l i g h t l y l e s s so. moderately  Two  f r o m salmon v i s c e r a b e i n g  good s o u r c e s o f r i b o f l a v i n .  I t was  suggested,  The  meal  prepared of the  because of t h e i r h i g h  c o n t e n t , t h a t s t i c k w a t e r meal and by-products  only  samples o f salmon r o e m e a l s were o n l y  f r o m s a l m o n heads had t h e l o w e s t v i t a m i n G p o t e n c y series.  good  c e r t a i n of the  riboflavin other  under i n v e s t i g a t i o n s h o u l d be u t i l i z e d  i n the  manufacture of f i s h m e a l s . S e r i e s 2,  a l s o c o n s i s t i n g of three experiments,  s e v e n t y - f o u r l o t s of twenty-two c h i c k s each. a result  of t h i s  as e f f e c t i v e natural  shown as  s e r i e s t h a t s y n t h e t i c r i b o f l a v i n was  i n promoting  g r o w t h as was  equally  the r i b o f l a v i n  from  sources.  Contrary to previous practice, t o add  I t was  involved  i t was  not  found  necessary  d r i e d s k i m m i l k when f i s h m e a l s were s u p p l e m e n t e d w i t h  sufficient  vitamin  I t was  G.  found t h a t 3 7 0 micrograms of r i b o f l a v i n per 1 0 0  grams o f f e e d were a d e q u a t e f o r o p t i m a l g r o w t h and p r e v e n t i o n of c u r l e d - t o e p a r a l y s i s . requirements  The  minimum  f o r the riboflavin  f o r s a t i s f a c t o r y g r o w t h were much l o w e r  2 2 5 m i c r o g r a m s per 1 0 0 grams.  - namely,  - 44 REFERENCES  (1)  A c k e r s o n , C.W. , M.J. B l i s h and F.E. M u s s e h l 1935 The u t i l i z a t i o n o f n i t r o g e n , c a l c i u m , and p h o s p h o r u s by t h e g r o w i n g c h i c k . Nebraska E x p t . S t a . Res. B u i , 80,  (2)  A l m q u i s t , H.J. 1941 Chemical estimation of q u a l i t y i n animal p r o t e i n concentrates. J . 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C, Rose 1937 The r e l a t i o n o f c y s t i n e and m e t h i o n i n e t o g r o w t h . J o u r . B i o l . Chem. 121: No. 2: 403-410.  -  48  -  A i m q u i s t , H.J., E.L.R. S t o k s t a d a n d E.L. H a l b r o o k 1935 Values of animal p r o t e i n concentrates i n chick r a t i o n s . J o u r . N u t r . 1 0 : 19 3. The c o m p o s i t i o n and n u t r i t i v e p r o p e r t i e s o f soybeans a n d s o y b e a n o i l m e a l . - A L i t e r a t u r e R e v i e w . 1939 Soybean N u t r i t i o n a l Research C o u n c i l , 3 8 1 8 Board o f Trade - B u i l d i n g , Chicago, 111. C s o n k a , F.A. and D.B. J o n e s 1934 The c y s t i n e , t r y p t o p h a n e and t y r o s i n e c o n t e n t o f t h e s o y b e a n . J o u r . A g r . R e s . 49 :  "279.  C s o n k a , F.A., and D.B. J o n e s 1 9 3 3 Differences i n the amino a c i d c o n t e n t o f t h e c h i e f p r o t e i n ( g l y c i n i n ) f r o m seeds o f s e v e r a l v a r i e t i e s o f s o y b e a n . Jour. Agr. Res. 4.6: 5 1 . C u l t o n , T.G., and H.R, B i r d 1940 The r i b o f l a v i n c o n t e n t o p o u l t r y f e e d s t u f f s . P o u l t r y S c i . 20: No. 1: 3-6. Hayward, J.¥., J . G. H a l p i n , C.E. Holmes, 0. B o h s t e d t and E.B. H a r t 19 3 7 Soybean o i l m e a l p r e p a r e d a t d i f f e r e n t temperatures as a f e e d f o r p o u l t r y . P o u l t r y S c i . 16:3-14 Hayward, J.W., H. S t e e n b o c k and G, B o h s t e d t 1 9 3 6 The e f f e c t o f c y s t i n e and c a s e i n s u p p l e m e n t s upon t h e n u t r i t i v e v a l u e o f t h e p r o t e i n o f raw and h e a t e d s o y b e a n s Jour. Nutr. 1 2 : 2 7 5 - 2 8 3 . H e n r y , W.A., and F.B. M o r r i s o n 2 0 t h e d . The M o r r i s o n P u b l i s h i n g  Feeds and F e e d i n g . Co,, I t h a c a , N.Y.  L l o y d , E.A., and J a c o b B i e l y 1941 P r a c t i c a l P o u l t r y F e e d i n g . P r o v . o f B.C. D e p t • o f A g r . B u i . 1 0 7 , V i c t o r i a , B.C. M i l l e r , David 1 9 3 8 The v i t a m i n s o f t h e B-G c o m p l e x . P o u l t r y S c i . 17, No 6 : 5 2 3 - 5 3 8 . e  Riboflavin. Annotated Rahway, N . J .  Bibliography.  1941 M e r c k &„Co. I n c .  R o b e r t s o n , E . I . , J . S . C a r v e r and J.W. Cook 1 9 3 8 Gross p r o t e i n v a l u e s o f some c o n c e n t r a t e s . P o u l t r y S c i . 1 7 : 433.  R o b e r t s o n , E . I . , J . S . C a r v e r , B.B. B o h r e n and J.W. Cook 1939 G r o s s v a l u e s o f c o m b i n a t i o n s o f some p r o t e i n concentrates. P o u l t r y S c i . 18: 410»  - 49 -  R o s e , • Wm. C., and E l d o n E. R i c e 1939 The u t i l i z a t i o n o f c e r t a i n s u l p h u r - c o n t a i n i n g compounds f o r g r o w t h p u r p o s e s . J o u r . B i o l . Chem. 1 3 0 : No. 1 : 3 0 3 - 3 2 3 . Sherwood, F.W., and J.O. H a l v e r s o n 1939 The v i t a m i n G c o n t e n t o f some o i l p r e s s - c a k e meals and r e l a t e d p r o d u c t s . . J o u r . A g r . R e s . 58, No. 10: 787-794. W i l g u s , H.S. , J r . , L.C. N o r r i s , and G.F. H e u s e r 1935 Haddock m e a l . E f f e c t o f m a n u f a c t u r i n g p r o c e s s upon • n u t r i t i v e v a l u e s . I n d . and E n g . Chem. 21'419-422.  

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