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Studies of the effects of different treatments on the metabolizable energy value of wheat El-Lakany, Safaa 1968

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STUDIES OF THE EFFECTS OF DIFFERENT TREATMENTS ON THE METABOLIZABLE ENERGY VALUE OF WHEAT by SAFAA EL-LAKANY A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE IN AGRICULTURE i n t h e Department o f P o u l t r y S c i e n c e We a c c e p t t h i s t h e s i s as c o n f o r m i n g t o t h e r e q u i r e d S t a n d a r d THE UNIVERSITY OF BRITISH COLUMBIA A p r i l 1968 In p r e s e n t i n g t h i s t h e s i s i n p a r t i a l f u l f i l m e n t o f t h e r e q u i r e m e n t s f o r an a d v a n c e d d e g r e e a t t h e 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 , I a g r e e t h a t t h e L i b r a r y s h a l l make i t f r e e l y a v a i l a b l e f o r r e f e r e n c e a nd S t u d y . I f u r t h e r a g r e e t h a t p e r m i s s i o n f o r e x t e n s i v e c o p y i n g o f t h i s t h e s i s f o r s c h o l a r l y p u r p o s e s may be g r a n t e d b y t h e Head o f my D e p a r t m e n t o r b y hits r e p r e s e n t a t i v e s . I t i s u n d e r s t o o d t h a t c o p y i n g o r p u b l i c a t i o n o f t h i s t h e s i s f o r f i n a n c i a l g a i n s h a l l n o t be a l l o w e d w i t h o u t my w r i t t e n p e r m i s s i o n . D e p a r t m e n t o f P „ u l ^ S c i , ^ f > 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 V a n c o u v e r 8, C a n a d a D a t e /| >a ( U Z J ABSTRACT F o u r s u c c e s s i v e e x p e r i m e n t s were c a r r i e d out t o s t u d y t h e e f f e c t s o f d i f f e r e n t t r e a t m e n t s on t h e n u t r i t i v e q u a l i t y o f 4 samples o f wheat. The t r e a t m e n t s used were a u t o c l a v i n g , f r e e z i n g , w a t e r - s o a k i n g , a c i d - s o a k i n g , b o i l i n g i n w a t e r , and c o m b i n a t i o n s o f t h e s e t r e a t m e n t s . M e t a b o l i z a b l e energy v a l u e s o f t h e v a r i o u s samples o f t r e a t e d and u n t r e a t e d wheat were d e t e r m i n e d u s i n g 2 - 3 weeks o l d c h i c k e n s . N i t r o g e n r e t e n t i o n f r o m d i e t s c o n t a i n i n g t h e d i f f e r e n t wheat samples was a l s o measured. A u t o c l a v i n g o f wheat samples f o r 60 m i n . a t 15 l b p r e s s u r e m a r k e d l y i n c r e a s e d t h e n u t r i t i v e q u a l i t y o f wheat as measured by m e t a b o l i z a b l e energy v a l u e s . F r e e z i n g a t -4°C f o r 5 days r e s u l t e d i n s i g n i f i c a n t improvement i n t h e m e t a b o l i z a b l e e nergy v a l u e o f t h e samples o f wheat t r e a t e d i n t h i s manner. Water- o r a c i d - s o a k i n g d i d n o t improve t h e m e t a b o l i z a b l e e n ergy v a l u e s o f t h e wheat samples. The d i f f e r e n t t r e a t m e n t s d i d n o t a f f e c t t h e p e r c e n t a g e n i t r o g e n r e t e n t i o n f r o m d i e t s c o n t a i n i n g t h e t r e a t e d wheat. P r o l o n g e d h e a t i n g ( a u t o c l a v i n g f o r 90 m i n . and 120 m i n . a t 15 l b p r e s s u r e ) r e d u c e d t h e energy v a l u e s o f wheat as w e l l as t h e p e r c e n t a g e n i t r o g e n r e t e n t i o n . TABLE OF CONTENTS Page I n t r o d u c t i o n and Review o f L i t e r a t u r e 1 M a t e r i a l s and Methods 14 R e s u l t s 20 E x p e r i m e n t 1 20 E x p e r i m e n t 2 20 E x p e r i m e n t 3 26 E x p e r i m e n t 4 31 D i s c u s s i o n 37 R e f e r e n c e s 45 ACKNOWLEDGEMENT The w r i t e r w i s h e s t o e x p r e s s h e r deep a p p r e c i a t i o n t o P r o f e s s o r Jacob B i e l y , Head o f t h e Department o f P o u l t r y S c i e n c e , and P r o f e s s o r B. E. March, o f t h e Department o f P o u l t r y S c i e n c e , f o r s u g g e s t i n g t h e pro b l e m , s u p e r v i s i n g t h e work, t h e i r encouragement and i n t e r e s t and o f f e r i n g a l l t h e f a c i l i t i e s needed f o r t h e c o m p l e t i o n o f t h i s s t u d y . G r a t e f u l acknowledgement i s a l s o e x t e n d e d t o P r o f e s s o r E. L. Watson o f t h e Department o f A g r i c u l t u r a l M e c h a n i c s , Dr. A. J . Renney P r o f e s s o r o f Agronomy and t o Dr. J . F. R i c h a r d s o f t h e Department o f P o u l t r y S c i e n c e f o r t h e i r v a l u a b l e a d v i c e and r e a d i n g t h e m a n u s c r i p t . INTRODUCTION AND REVIEW OF LITERATURE Wheat, t h e c u l t i v a t i o n o f w h i c h p r o b a b l y d a t e s back t o p r e - h i s t o r i c d a y s , i s one o f t h e most i m p o r t a n t o f the c e r e a l s . F o r t u n a t e l y , i t may be grown under a wide v a r i e t y o f s o i l and c l i m a t i c c o n d i t i o n s . Thus i t i s one o f t h e most w i d e l y grown c r o p s i n the w o r l d . Numerous v a r i e t i e s o f wheat a r e known and P e l s h e n k e (1930) has s t a t e d t h a t 12,000 v a r i e t i e s o f wheat have been i d e n t i f i e d t h r o u g h -o u t t h e w o r l d . Other a u t h o r i t i e s g i v e the t o t a l as something l i k e 30,000. A l l wheat, whether w i l d o r c u l t i v a t e d , b e l o n g s t o t h e genus T r i t i c u m o f t h e f a m i l y Gramineae, t h e " g r a s s f a m i l y " . T r i t i c u m i s o n l y one o f some 600 genera b e l o n g i n g t o t h i s g r e a t f a m i l y , w h i c h i t s e l f c o m p r i s e s w e l l o v e r 5,000 s p e c i e s . The wheat g r a i n i s used f o r human co n s u m p t i o n , f o r l i v e s t o c k f e e d , f o r seed and f o r i n d u s t r i a l p u r p o s e s . Malenbaum (1 9 5 3 ) , i n a s t u d y c o v e r i n g t h e f i v e y e a r s p r e c e d i n g The Second World War, made the f o l l o w i n g e s t i m a t e s o f t h e u t i l i z a t i o n o f wheat: 1. 74 p e r c e n t f o r human co n s u m p t i o n , 2. 11 p e r c e n t used f o r p r o p a g a t i o n , 3. 15 p e r -c e n t f e d t o l i v e s t o c k , u t i l i z e d i n i n d u s t r y , o r waste d . The wheat k e r n e l may be r e g a r d e d as c o n s i s t i n g o f t h r e e p a r t s : 1. The germ o r embryo i n c l u d i n g i t s s h e a t h , the s c u t e l l u m w h i c h produces t h e new p l a n t . 2. The s t a r c h y endosperm. 3. The v a r i o u s o u t e r c o v e r i n g s c o n s t i t u t i n g t h e husk o f t h e g r a i n . A c c o r d i n g t o K e n t - J o n e s and Amos (1950) t h e average c o m p o s i t i o n o f a wheat k e r n e l i s a p p r o x i m a t e l y 85% endosperm, 2% embryo and 137. husk. L a t e r (1957) t h e same a u t h o r s , f r o m t h e i r own e x t e n s i v e d a t a and f r o m d a t a o f o t h e r i n v e s t i g a t o r s , have p r e s e n t e d t a b l e s showing t h e a p p r o x i m a t e c h e m i c a l c o m p o s i t i o n o f wheats from d i f f e r e n t p a r t s o f t h e w o r l d and o f t h e v a r i o u s p r o d u c t s o f m i l l i n g d e r i v e d f r o m s u c h wheats. The c a r b o h y d r a t e s found i n wheat i n c l u d e s t a r c h , c e l l u l o s e and s u g a r s . S t a r c h i s t h e most abundant c o n s t i t u e n t o f t h e wheat k e r n e l and i s c o n f i n e d a l m o s t e n t i r e l y t o t h e endosperm. The s t a r c h i s i m p o r t a n t i n a n i m a l n u t r i t i o n c h i e f l y as an e f f e c t i v e and i n e x p e n s i v e s o u r c e of energy. Wheat i s a s o u r c e , n o t o n l y o f c a r b o h y d r a t e s b u t a l s o o f p r o t e i n , s u p p l y i n g adequate amounts o f some, b u t n o t a l l , o f t h e e s s e n t i a l amino a c i d s . Thus whole wheat i s a good s o u r c e o f i s o l e u c i n e , l e u c i n e , t h r e o n i n e , and v a l i n e , b u t o n l y a f a i r s o u r c e o f l y s i n e , m e t h i o n i n e , p h e n y l a l a n i n e , and t r y p t o p h a n . These e i g h t e s s e n t i a l amino a c i d s a r e found i n wheat f l o u r i n s i m i l a r p r o p o r t i o n s b u t i n lo w e r o v e r a l l c o n c e n t r a t i o n . The main d e f i c i e n c i e s a r e i n t h e amounts o f l y s i n e , t r y p t o p h a n , and m e t h i o n i n e . The f a t s o f t h e wheat g r a i n a r e o b v i o u s l y n o t p r e s e n t i n s u f f i c i e n t q u a n t i t i e s t o p l a y an i m p o r t a n t r o l e as s o u r c e s o f energy. However, whole wheat i s an e x c e l l e n t s o u r c e o f t h e f a t - s o l u b l e v i t a m i n E. The o t h e r f a t - s o l u b l e v i t a m i n s A, D and K a r e n o t f o u n d i n wheat. As r e g a r d s w a t e r - s o l u b l e v i t a m i n s , wheat c o n t a i n s s i g n i f i c a n t amounts o f 3 most o f t h e B - v i t a m i n s b u t e s p e c i a l l y t h i a m i n e . Whole wheat s u p p l i e s s i g n i f i c a n t amounts o f i r o n , phosphorus i n t h e form o f p h y t i n p h o s p h o r u s , magnesium, manganese, copper and z i n c . I t i s e v i d e n t from t h e f o r e g o i n g b r i e f s u r v e y t h a t wheat has much t o o f f e r as a b a s i c f o o d , b u t t h a t , l i k e a l l o t h e r f o o d s , i t can n o t f o r m t h e s o l e s o u r c e o f n u t r i e n t s f o r a n i m a l s o r humans. S e v e r a l i n v e s t i g a t o r s have r e p o r t e d t h a t t h e use o f h e a t i n t h e p r e p a r a t i o n o f c e r t a i n f o ods d e f i n i t e l y a l t e r s 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 . Heat damage t o p r o t e i n becomes a t t i m e s a p r o b l e m o f c o n s i d e r a b l e n u t r i t i o n a l i m p o r t a n c e . McCollum and D a v i s (1915) r e p o r t e d a. l o s s o f n u t r i t i v e v a l u e i n c a s e i n a u t o c l a v e d f o r one hour a t 15 l b p r e s s u r e and showed t h a t t h i s l o s s was due t o changes i n t h e p r o t e i n f r a c t i o n and not t o t o x i c i t y . G e i l i n g , i n 1917, c o r r o b o r a t e d t h e s e f i n d i n g s . I n 1925, Morgan and K i n g showed t h a t h e a t i n g o f c e r t a i n c e r e a l f o o d s caused a n u t r i t i v e i m p a i r m e n t , as e v i d e n c e d by a r e t a r d a t i o n o f growth i n r a t s f e d t o a s t e d c r a c k e d whole wheat, t o a s t e d w h i t e b r e a d , c r u s t o f w h i t e b r e a d , and p u f f e d wheat o r p u f f e d r i c e . S h o r t l y t h e r e a f t e r Morgan and h e r a s s o c i a t e s (1931) found t h a t d r y h e a t a t a p p r o x i m a t e l y 200°C f o r 45 min. caused a l o s s o f much o f t h e growth p r o m o t i n g v a l u e o f t h e c e r e a l f o ods t e s t e d , b u t s u p p l e m e n t a t i o n w i t h c a s e i n (57.) v e r y n e a r l y compensated f o r t h e l o s s i n n u t r i t i v e v a l u e . Because d i g e s t i b i l i t y o f th e t o a s t e d p r o t e i n s as compared w i t h t h e raw c e r e a l s was b u t l i t t l e a l t e r e d , t h o s e w o r k e r s c o n c l u d e d t h a t t h e e f f e c t s o f t h e h e a t t r e a t m e n t 4 were p r o b a b l y r e l a t e d i n some way t o a l t e r a t i o n o f t h e p a t t e r n . o f amino a c i d s a v a i l a b l e f o r a b s o r p t i o n . I n an at t e m p t t o l o c a t e c h e m i c a l l y t h e s i t e o f p r o t e i n i n j u r y a f f e c t e d by h e a t i n g , B l o c k ejt a l . (1934) c a l l e d a t t e n t i o n t o t h e p a r t i c u l a r v u l n e r a b i l i t y o f l y s i n e , f o r example, a f t e r h e a t i n g c a s e i n a t 150°C f o r 65 min., t h e y r e c o v e r e d p r a c t i c a l l y as much l y s i n e from t h e h e a t e d c a s e i n as f r o m unheated c a s e i n , t h u s p r o v i n g t h a t t h e l y s i n e had n o t been d e s t r o y e d by t h e h e a t i n g . They c o n c l u d e d , t h e r e f o r e t h a t t h e mechanism o f h e a t i n j u r y i s r e l a t e d i n some way t o the f a c t t h a t t h e l y s i n e i s made u n a v a i l a b l e f o r n u t r i t i v e p u r p o s e s . They p o s t u l a t e d a m o l e c u l a r r e-arrangement whereby a p a r t o f t h e l y s i n e p r e c u r s o r s became r e s i s t a n t t o e n z y m a t i c d e g r a d a t i o n , and a t t r i b u t e d t h e l o w e r e d n u t r i t i v e v a l u e o f the h e a t e d p r o t e i n t o i t s r e s i s t a n c e t o e n z y m a t i c h y d r o l y s i s . C h i c k e_t a l . i n 1935, by means o f n i t r o g e n b a l a n c e t e c h n i q u e s i n a d u l t r a t s , d e m o n s t r a t e d a r e d u c t i o n o f b i o l o g i c a l v a l u e i n c a s e i n s u b j e c t e d t o d r y h e a t a t 150°C f o r 66 h o u r s b u t n o t when h e a t e d b e l o w 125°C, and i n l a c t a l b u m i n h e a t e d a t 120°C f o r 72 h o u r s . T h e r e f o r e , i t may be s a i d t h a t i t i s e v i d e n t t h a t s e v e r a l t y p e s o f p r o t e i n s were s u s c e p t i b l e t o h e a t i n j u r y a t v a r y i n g d e grees o f t e m p e r a t u r e and t i m e o f e x p o s u r e . Moreover t h e r e were i n d i c a t i o n s t h a t t h e mechanism o f i n j u r y was r e l a t e d i n some way t o amino a c i d i n t e r a c t i o n s . L a t e r a t t e n t i o n has been f o c u s e d upon t h e n o n - e n z y m a t i c browning r e a c t i o n as a p o s s i b l e cause o f h e a t i n j u r y . T h i s r e a c t i o n , f i r s t d e s c r i b e d by M a i l l a r d i n 1912, i s t h e consequence o f an i n t e r a c t i o n between amino a c i d s and r e d u c i n g s u g a r s under t h e i n f l u e n c e o f h e a t . 5 H a l e n y and GuggenMum (1953) d e m o n s t r a t e d t h a t a u t o c l a v i n g o f g l u t e n i n t h e p r e s e n c e o f g l u c o s e r e s u l t s i n a c o n s i d e r a b l e impairment o f i t s n u t r i t i v e v a l u e . T h i s e f f e c t seems t o be due t o a d i m i n i s h e d e n z y m a t i c l i b e r a t i o n o f t h e c o n s t i t u e n t e s s e n t i a l amino a c i d s and thus t o a d e c r e a s e i n the a v a i l a b i l i t y o f t h e s e amino a c i d s . A r g i n i n e and l y s i n e appear t o be most a f f e c t e d and t h e r e f o r e l i m i t t h e b i o l o g i c a l v a l u e o f the p r o t e i n . They s u g g e s t t h a t on h e a t i n g g l u t e n w i t h g l u c o s e , an e n z y m e - r e s i s t a n t a c i d n o n - r e s i s t a n t l i n k a g e f o r m between a r g i n i n e and l y s i n e , r e s p e c t i v e l y , and g l u c o s e . These r e a c t i o n s were f o l l o w e d upon f u r t h e r h e a t i n g by t h e f o r m a t i o n o f an e n z y m a t i c -and a c i d - r e s i s t a n t l i n k a g e o r by t h e d i s i n t e g r a t i o n o f t h e s e amino a c i d m o l e c u l e s . Emphasis upon t h e d e l e t e r i o u s a s p e c t s o f s e v e r e h e a t t r e a t m e n t o f p r o t e i n s h o u l d n o t cause the f a c t t o be o v e r l o o k e d t h a t i n t h e moderate t e m p e r a t u r e ranges t h e r e a r e no s u c h a d v e r s e e f f e c t s . I n f a c t , as has been shown r e p e a t e d l y w i t h r e s p e c t t o soybean p r o t e i n , moderate h e a t i n g may a c t u a l l y improve n u t r i t i v e v a l u e . Thus Hayward e t a l . (1936) d e m o n s t r a t e d t h a t h e a t i n g o f soybean meal caused n u t r i t i v e improvement. F r i t z e t a l . (1947) found t h a t ground soybeans a u t o c l a v e d a t 15 l b f o r 20 - 30 m i n . r e s u l t e d i n a good b i o l o g i c a l v a l u e when f e d t o p o u l t s . P a r s o n s and h e r a s s o c i a t e s (1939) o b s e r v e d t h a t a u t o c l a v i n g o f soybeans improved the n u t r i t i v e v a l u e up t o a p o i n t , beyond w h i c h d e t e r i o r a t i o n o c c u r r e d a f t e r exposure a t h i g h e r t e m p e r a t u r e f o r l o n g e r p e r i o d s . I n t h e l a t t e r i n s t a n c e , l y s i n e s u p p l e m e n t a t i o n f a i l e d t o g i v e improvement. 6 Evans e_t al. (1947) o b t a i n e d b e t t e r c h i c k growth w i t h soybean meal a u t o c l a v e d between 100° and 120°C f o r 30 min. t h a n w i t h soybean meal a u t o c l a v e d a t 130°C. The meal a u t o c l a v e d a t the lower t e m p e r a t u r e was more c o m p l e t e l y d i g e s t e d by the c h i c k s and a l s o by t r y p s i n and e r e p s i n in v i t r o . Other i n v e s t i g a t i o n s have g i v e n c o n s i d e r a t i o n t o t h e r e m o v a l o f h y p o t h e t i c a l t o x i c f a c t o r s by h e a t d e s t r u c t i o n . S h r e w s b e r r y and c o w o r k e r s (1932) o b s e r v e d t h a t c o o k i n g i n c r e a s e d the d i g e s t i b i l i t y o f t h e soybeans o n l y s l i g h t l y . T h e i r t h e o r y was t h a t h e a t i n g caused t h e r e m o v a l o r d e s t r u c t i o n o f c e r t a i n m a t e r i a l s o f a t o x i c n a t u r e . A t r y p s i n i n h i b i t o r has been shown t o be p r e s e n t i n raw soybean o i l meal (Ham, S a n d s t e d t i n 1 9 4 4 ) . Bowman (1946) p r e s e n t e d e v i d e n c e t h a t more t h a n one t r y p s i n i n h i b i t o r i s p r e s e n t i n s o y b e a n s . R a c h i s and A n d e r s o n (1965) i s o l a t e d f o u r t r y p s i n i n h i b i t o r s f r o m soybean by d i e t h y l a m i n o e t h y l -c e l l u l o s e chromatography. I n wheat, as w e l l , t h e b e n e f i c i a l e f f e c t o f m o i s t h e a t on t h e n u t r i t i v e v a l u e has been p a r t i a l l y e s t a b l i s h e d . O b s e r v a t i o n s as t o the e f f e c t o f r a i s i n g wheat and f l o u r t o r e l a t i v e l y h i g h t e m p e r a t u r e , a p p r o x i m a t e l y 150°F, were made many y e a r s ago. Thus Weyl and B i s c h o f f (1882) o b s e r v e d t h a t h e a t i n g f l o u r t o 140"F f o r s e v e r a l h o urs p r e v e n t e d subsequent w a s h i n g out o f t h e g l u t e n and t h i s was c o n f i r m e d by Jago ( 1 9 1 1 ) , who p o i n t e d out t h a t t h e heat had d e s t r o y e d t h e n ormal n a t u r e o f t h e g l u t e n . B e a u d o i n ejt al. d e m o n s t r a t e d i n 1951 t h e s i g n i f i c a n t improvement i n t h e b i o l o g i c a l v a l u e o f t h e p r o t e i n o f the whole wheat 7 when s u b j e c t e d t o c o o k i n g i n b o i l i n g w a t e r as i n t h e p r e p a r a t i o n o f s h redded wheat. P a r b o i l e d wheat, o r b u r g h u l , i s p r e p a r e d by s u b m i t t i n g whole g r a i n wheat t o b o i l i n g f o r about t h r e e h o u r s f o l l o w e d by r a p i d d r y i n g i n h o t sun. A c c o r d i n g t o Shammas and A d o l p h (1954) t h i s k i n d o f h e a t t r e a t m e n t does n o t a l t e r 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 f r a c t i o n measured i n terms o f t h e p r o t e i n e f f i c i e n c y i n r a t s . A s i m i l a r s t u d y has been c o n d u c t e d by Yang and A l - N o u r i (1962) t o compare t h e n u t r i t i o n a l v a l u e o f t h e p r o t e i n s o f p a r b o i l e d wheat, p r e p a r e d f r o m B r e r o r s o f t w h i t e wheat, w i t h t h a t o f whole g r a i n o r w h i t e f l o u r f r o m t h e same wheat. The p r o t e i n e f f i c i e n c y r a t i o s and b i o l o g i c a l v a l u e s o b t a i n e d w i t h t h e d i e t s c o n t a i n i n g p a r b o i l e d wheat as t h e s o l e s o u r c e o f p r o t e i n o r supplemented w i t h c a s e i n were no t d i f f e r e n t f r o m t h o s e of wheat g r a i n b u t were g r e a t e r t h a n t h o s e o f w h i t e f l o u r , under comparable c o n d i t i o n s . K o h l e r (1964) s t u d i e d t h e e f f e c t o f p r o c e s s i n g on t h e b i o l o g i c a l v a l u e o f wheat p r o t e i n s . A p p l i c a t i o n o f c h e m i c a l and b i o -l o g i c a l t e c h n i q u e s t o b u l g u r p r o c e s s i n g has shown t h a t t h e more s e v e r e p r o c e s s e s can a d v e r s e l y a f f e c t p r o t e i n q u a l i t y , b u t t h a t t h e m i l d e r , c o n t r o l l e d p r o c e s s e s a r e n o t h a r m f u l ( s t e e p i n g a t 80°C f o r 70 min. and c o o k i n g a t 5 p s i f o r 30 m i n . ) . A s e r i e s o f e x p e r i m e n t s was c o n d u c t e d by H a s k e l l e_t a l . (1961) t o s t u d y t h e e f f e c t s o f v a r i o u s t r e a t m e n t s on t h e n u t r i t i o n a l v a l u e o f b a r l e y and wheat and wheat mixed f e e d . They f o u n d t h a t a u t o c l a v i n g and m i l d a c i d h y d r o l y s i s o f t h e c e r e a l g r a i n s g r e a t l y 8 i n c r e a s e d c h i c k g r o w t h . A u t o c l a v i n g o n l y o f wheat and wheat mixed f e e d gave some improvement i n n u t r i t i v e v a l u e . R e s u l t s p r e s e n t e d by Adams e t aJL. (1961) showed t h a t t h e gr o w t h p r o m o t i n g e f f e c t due t o w a t e r s o a k i n g o f g r a i n s can be d u p l i c a t e d by a c i d s o a k i n g . Steam e x p a n s i o n causes a breakdown o f c e l l u l a r s t r u c t u r e i n t he ground g r a i n w i t h o u t p r o d u c i n g t h e w a t e r t r e a t m e n t r e s p o n s e . They found t h a t f e e d i n g h a r d wheat and r e c o n s t i t u t e d wheat i n w h i c h components o f t h i s g r a i n a r e w a t e r t r e a t e d shows t h a t t h e w a t e r t r e a t -ment e f f e c t s r e s i d u e s p r i m a r i l y i n t h e f l o u r p o r t i o n o f t h e g r a i n . A l t h o u g h c h i c k s f e d w a t e r - o r a c i d - t r e a t e d g r a i n grow a t a f a s t e r r a t e t h a n c h i c k s f e d u n t r e a t e d g r a i n t h e r e was no d i f f e r e n c e i n n i t r o g e n r e t e n t i o n . However, t h e m e t a b o l i z a b l e energy v a l u e o f the t r e a t e d r a t i o n s i n d i c a t e d t h a t w a t e r t r e a t m e n t a f f e c t e d energy u t i l i z a t i o n w i t h o u t a f f e c t i n g p r o t e i n u t i l i z a t i o n . S i m i l a r improvement i n f e e d i n g v a l u e o f wheat upon w a t e r -t r e a t m e n t has been a c h i e v e d by Leong e t al. ( 1 9 6 0 ) . They found t h a t t h e f e e d i n g v a l u e o f wheat was improved s i g n i f i c a n t l y (13 - 19%) by w a t e r - t r e a t m e n t and t h a t wheat so t r e a t e d was e q u a l t o o r b e t t e r t h a n c o r n , i n n u t r i t i o n a l v a l u e f o r p o u l t r y . C h i c k e x p e r i m e n t s w i t h wheat m i l l i n g f r a c t i o n s showed t h a t w a t e r - t r e a t m e n t improved t h e u t i l i z a t i o n o f a l l m i l l i n g f r a c t i o n s e x c e p t f l o u r . I n 1934 B o a s - F i x s e n et. aJL. o b s e r v e d t h a t w e a n l i n g r a t s grew a t a somewhat g r e a t e r r a t e on a steamed wheat d i e t t h a n on a raw wheat d i e t . I n t h o s e e x p e r i m e n t s t h e wheat d i e t was steamed f o r 40 min. a t a m o i s t u r e c o n t e n t o f about 50%. Such t r e a t m e n t would have g e l a t i n i z e d 9 t h e s t a r c h , and t h i s c o u l d have been a f a c t o r i n p r o m o t i n g t h e improved r a t e o f g r owth w h i c h was o b s e r v e d . H u t c h i n s o n e£ a l . (1964) compared t h e v a l u e o f m i l d l y steamed wheat i n p r o m o t i n g t h e growth o f t h e w e a n l i n g r a t w i t h t h a t o f unheated wheat. D i e t s c o n t a i n i n g f l o u r o r whole meal from t h e steamed wheat promoted a h i g h e r r a t e o f g r o w t h and o f f o o d c o n s u m p t i o n t h a n t h o s e c o n t a i n i n g c o r r e s p o n d i n g m a t e r i a l from unheated wheat. S e v e r a l y e a r s ago, w h i l e t r y i n g t o a s s e s s the a v a i l a b i l i t y o f manganese i n wheat germ m e a l , Creek (1955) n o t e d t h a t t h e meal was h i g h l y d e t r i m e n t a l t o the growth o f c h i c k . Subsequent work c o n f i r m e d t h i s f i n d i n g , and i t was d e t e r m i n e d t h a t t h e g rowth i n h i b i t i o n c o u l d be r e v e r s e d by a u t o c l a v i n g f o r 30 min. a t 121°C and 15 l b p r e s s u r e . S u b s e q u e n t l y Creek et_ a l . (1961) showed t h a t raw wheat germ meal d e p r e s s e s g r o w t h and f e e d e f f i c i e n c y b u t a u t o c l a v i n g improves b o t h g r o w t h and f e e d e f f i c i e n c y . T h i s i s i n d i c a t i v e , as t h e y p r o p o s e d , o f t h e p r e s e n c e o f a t h e r m o l a b i l e f a c t o r w h i c h i m p a i r s d i g e s t i o n . E v i d e n c e as t o t h e p r e s e n c e o f a t h e r m o l a b i l e g r o w t h i n h i b i t o r w h i c h a f f e c t e d t h e g rowth o f young c h i c k s has been p r e s e n t e d by Creek once a g a i n i n 1962. The i n h i b i t i n g f a c t o r seems s p e c i f i c a l l y t o b l o c k u t i l i z a t i o n o f f a t . However, Cr e e k et a l . f o u n d a l s o , t h a t n i t r o g e n r e t e n t i o n i s a l s o improved somewhat by h e a t i n g i n d i c a t i n g t h a t p r o t e i n a v a i l a b i l i t y might be i n c r e a s e d . Learmonth and Wood (1963) found t h a t aqueous e x t r a c t s o f wheat f l o u r i n h i b i t e d t h e p r o t e o l y t i c a c t i o n o f t r y p s i n on a g e l a t i o n s u b s t r a t e by means o f t h e g e l a t i n t e s t . I t i s a l s o n o t e d t h a t t h e i n h i b i t o r w h i c h i s p r e s e n t i n wheat f l o u r s from 10 w i d e l y d i f f e r e n t s o u r c e s , i s n o t a t t r i b u t a b l e t o c h e m i c a l t r e a t m e n t o r a d d i t i v e and i s n o t a s s o c i a t e d w i t h t h e b r a n and germ f r a c t i o n . C o o k i n g i s shown t o reduc e t h e i n h i b i t i n g a c t i v i t y . Shyamala e_t al. (1961) s t a t e d t h a t wheat c o n t a i n s a t r y p s i n i n h i b i t o r w h i c h i s s i m i l a r t o , but n o t as p o t e n t a s , i t s c o u n t e r p a r t f o u n d i n soybean. However, H u t c h i n s o n e_t al. (1960) found t h a t h e a t p r o c e s s i n g o f raw wheat germ gave s l i g h t l y l o w e r r a t e s o f g r o w t h and food c o n s u m p t i o n , when f e d t o w e a n l i n g r a t s . R e c e n t l y (1967) O l s e n s t u d i e d t h e e f f e c t o f h e a t t r e a t m e n t o f wheat germ meal on t h e q u a l i t y and b i o l o g i c a l a v a i l a b i l i t y o f t h e p r o t e i n f r a c t i o n . She f o u n d t h a t a r g i n i n e and l y s i n e were d e c r e a s e d by a l l t r e a t m e n t s and t h a t t h e g r e a t e s t l o s s was o f l y s i n e i n samples w h i c h had been a u t o c l a v e d f o r 45 and 90 min. D i g e s t i b i l i t y o f p r o t e i n and a b s o r p t i o n o f amino a c i d s , as d e t e r m i n e d w i t h r a t s , were d e c r e a s e d by a u t o c l a v i n g . D e c r e a s e d amino a c i d s a v a i l a b i l i t y and p r o t e i n d i g e s t -i b i l i t y were r e f l e c t e d i n l o w e r w e i g h t g a i n s and a r e d u c t i o n i n t h e p r o t e i n e f f i c i e n c y r a t i o . O l s e n s t a t e d t h a t raw wheat germ meal e x h i b i t e d no t o x i c i t y f o r t h e r a t s and s u g g e s t e d t h a t h e a t t r e a t m e n t a p p l i e d d u r i n g p r o c e s s i n g s h o u l d be k e p t at a minimum. H i g h p r e s s u r e p e l l e t i n g w i t h steam t r e a t m e n t has f r e q u e n t l y been shown t o i n c r e a s e i n t a k e o f t h e f e e d , and so enhance r a t e o f g r o w t h (Haywang and Morgan, 1944). T h i s improvement i n w e i g h t g a i n i s u s u a l l y g r e a t e r f o r f e e d s o f h i g h e r f i b e r c o n t e n t ( S l i n g e r e t a l . , 11 1949; B e a r s e e t a l . , 1952; Dymsza e t a l . , 1955). S i n c e p e l l e t i n g has produced l i t t l e improvement o f r a t i o n s h i g h i n f a t c o n t e n t (Pepper e t a l . , 1960; A l l e r d e t a l . , 1957; Combs, 1959) i t has been s u g g e s t e d t h a t t h e b e n e f i c i a l e f f e c t i s a r e s u l t o f i n c r e a s e d d e n s i t y r a i s i n g t h e i n t a k e o f n u t r i e n t s . However, f u r t h e r e x p e r i m e n t a l work, i n w h i c h f e e d i n t a k e was c o n t r o l l e d , i n d i c a t e d t h a t g r e a t e r i n g e s t i o n o f p e l l e t e d f e e d i s not s o l e l y r e s p o n s i b l e ( B l a c k £t a l . , 1958), a l s o where r e g r o u n d p e l l e t s have been f e d t o c h i c k s t h e r e i s an i n d i c a t i o n t h a t p e l l e t i n g has more t h a n a p h y s i c a l e f f e c t i n e n h a n c i n g c h i c k g r o w t h ( A l l e r d e t a l . , 1957). A l l e r d ejt a l . p r o p o s e d t h a t t h e r e i s a c h e m i c a l e f f e c t , due t o p e l l e t i n g a t h i g h t e m p e r a t u r e and p r e s s u r e , w h i c h e i t h e r improves t h e a v a i l a b i l i t y o f some o f t h e n u t r i e n t s o r d e s t r o y s a t o x i c f a c t o r p r e s e n t i n f e e d . No d i f f e r e n c e s i n c h e m i c a l c o m p o s i t i o n between p e l l e t e d and n o n - p e l l e t e d f e e d were found by B o l t o n , ( 1 9 6 0 ) . However, t h e e f f e c t s o f g r i n d i n g , p e l l e t i n g and d i e t b a l a n c e on t h e m e t a b o l i z a b l e energy v a l u e s o f c e r e a l g r a i n s and on t h e performance o f c h i c k e n s have been r e v i e w e d by M c i n t o s h ( 1 9 6 2 ) . The r e s u l t s o b t a i n e d by t h e s e w o r k e r s i n d i c a t e d t h a t no c o n s i s t e n t i n c r e a s e i n m e t a b o l i z a b l e e nergy r e s u l t e d f r o m t h e g r i n d i n g o r p e l l e t i n g o f c e r e a l g r a i n . Whole wheat was. found t o y i e l d more m e t a b o l i z a b l e energy t h a n ground (two e x p e r i m e n t s ) o r p e l l e t e d (one e x p e r i m e n t ) f o r m s . However, i n a t h i r d e x p e r i m e n t no d i f f e r e n c e was e v i d e n t i n t h e m e t a b o l i z a b l e energy c o n t e n t o f t h e d i e t s o r i n t h e w e i g h t g a i n s o r i n t h e e f f i c i e n c y o f f e e d c o n v e r s i o n by c h i c k e n s f e d ground, p e l l e t e d o r whole wheat. 12 S t u d i e s by J e n s e n (1961) on t h e b a s i s o f t h e growth r e s p o n s e o f b i r d s f e d p e l l e t s have l e d t o the c o n c l u s i o n t h a t p e l l e t i n g i n c r e a s e s t h e a v a i l a b l e energy o f a r a t i o n . The m e t a b o l i z a b l e e nergy c o n t e n t o f p e l l e t e d r a t i o n s was found t o be s l i g h t l y h i g h e r t h a n t h a t o f u n p e l l e t e d r a t i o n s . Gave e_t al. (1965) found t h a t h i g h - p r e s s u r e s t e a m - p e l l e t i n g o f r a t i o n s c o n t a i n i n g 507» o f wheat b y - p r o d u c t s improved g r o w t h and f e e d e f f i c i e n c y m a r k e d l y . The l e v e l o f f e e d i n t a k e was i n c r e a s e d f o r a l l r a t i o n s and t h e m e t a b o l i z a b l e energy v a l u e o f b r a n , s h o r t s and wheat germ meal were i n c r e a s e d by 30, 14 and 157. r e s p e c t i v e l y . S i n c e most p r e v i o u s work has used r a t e o f g r o w t h and i t s e f f i c i e n c y i n r e l a t i o n t o p r o t e i n i n t a k e as a c r i t e r i a o f n u t r i t i o n a l v a l u e , i t i s i m p o r t a n t t o know whether the h e a t i n g c o n d i t i o n s w h i c h a r e o p t i m a l a c c o r d i n g t o t h e s e c r i t e r i a a r e a l s o o p t i m a l f o r g r e a t e s t u t i l i z -a t i o n as measured by m e t a b o l i z a b l e e n e r g y . Renner and H i l l (1960) a p p l i e d t h e m e t a b o l i z a b l e energy method t o t h e q u a n t i t a t i v e e v a l u a t i o n o f t h e e f f e c t s o f h e a t i n g on soybeans and e x t r a c t e d soybean f l a k e s as a measure o f o v e r a l l u t i l i z a t i o n . They s t a t e d t h a t t h e degree o f heat t r e a t m e n t w h i c h produced maximal m e t a b o l i z -a b l e energy v a l u e a l s o produced maximal r a t e o f g r o w t h . The c o n d i t i o n s t h e y used were 107°C f o r 10 - 60 m i n . f o r e x t r a c t e d f l a k e s , and 10 min. a t 107°C f o r ground soybeans. O t h e r s t u d i e s by t h e same a u t h o r s were u n d e r t a k e n t o d e t e r m i n e the r e l a t i o n s h i p s between h e a t t r e a t m e n t and u t i l i z a t i o n o f ground soybean and e x t r a c t e d d e h u l l e d soybean f l a k e s by measurement o f 13 m e t a b o l i z a b l e energy and f a t a b s o r b a b i l i t y . I t was o b s e r v e d t h a t m e t a b o l i z a b l e energy v a l u e s o f t h e unheated m a t e r i a l s were m a r k e d l y and s i g n i f i c a n t l y l o w e r t h a n t h o s e o f the o p t i m a l l y h e a t e d samples. They a l s o f ound t h a t a b s o r b a b i l i t y o f d i e t a r y f a t was i m p a i r e d by unheated samples. The m e t a b o l i z a b l e energy v a l u e s o f wheat and wheat b y - p r o d u c t s have been s t u d i e d by H i l l e t a l . ( 1 9 6 0 ) , P o t t e r ( 1 9 6 0 ) , S i b b a l d and S l i n g e r ( 1 9 6 2 ) , S i b b a l d and S l i n g e r (1963) and Schumaier and M c G i n n i s ( 1 9 6 7 ) . No i n f o r m a t i o n i s a v a i l a b l e i n t h e l i t e r a t u r e upon the e f f e c t o f a u t o c l a v i n g o f whole wheat upon t h e m e t a b o l i z a b l e energy v a l u e s . The f o l l o w i n g s t u d y u n d e r t a k e s t o examine t h i s a s p e c t o f n u t r i t i o n . 14 MATERIALS AND METHODS F o u r s u c c e s s i v e e x p e r i m e n t s were c o n d u c t e d t o d e t e r m i n e t h e e f f e c t o f d i f f e r e n t t r e a t m e n t s on t h e m e t a b o l i z a b l e energy v a l u e s o f whole wheat. I . G e n e r a l D a y - o l d White L e g h o r n c o c k e r e l c h i c k s were used as e x p e r i m e n t a l s u b j e c t s . The c h i c k s were r e a r e d i n e l e c t r i c a l l y h e a t e d , w i r e f l o o r e d pens. They were g i v e n w a t e r and f e e d ad l i b i t u m d u r i n g t h e p r e l i m i n a r y p e r i o d , w h i c h l a s t e d two weeks f o r e x p e r i m e n t s 1, 3, and 4 and 3 weeks f o r e x p e r i m e n t 2. C o m p o s i t i o n o f t h e b a s a l d i e t f e d d u r i n g t h e p r e l i m i n a r y p e r i o d i s shown i n T a b l e 1. The average n i t r o g e n c o n t e n t and m e t a b o l i z a b l e energy v a l u e f o r t h e b a s a l d i e t were 3.927» and 3378.4 c a l / g r a m d r y m a t t e r , r e s p e c t i v e l y . Three r e p l i c a t e pens were a s s i g n e d t o each o f t h e d i e t a r y t r e a t m e n t s . The number o f b i r d s p e r r e p l i c a t e was 8 i n the f i r s t e x p e r i -ment and 6 i n t h e subsequent e x p e r i m e n t s . E x p e r i m e n t a l d i e t s were f o r m u l a t e d by a d d i n g , on a d r y m a t t e r b a s i s , the t e s t i n g r e d i e n t s a t t h e expense o f a p o r t i o n o f t h e t o t a l b a s a l d i e t . The r a t e o f s u b s t i t u t i o n was 257. o f t h e d i e t . The c h i c k s were f e d t h e e x p e r i m e n t a l d i e t s f o r one day t o accustom them t o t h e d i e t s . Feed i n t a k e was measured and e x c r e t a was c o l l e c t e d o v e r t h e f o l l o w i n g 3-day p e r i o d . The e x c r e t a samples were f r e e z e - d r i e d and ground. The d i e t s and t h e d r i e d e x c r e t a were a s s a y e d f o r g r o s s energy u s i n g a P a r r Oxygen TABLE 1. C o m p o s i t i o n o f b a s a l d i e t Component Ground wheat Ground y e l l o w c o r n Soybean meal 487. p r o t e i n H e r r i n g meal 72% p r o t e i n D r i e d d i s t i l l e r s ' s o l u b l e s D e h y d r a t e d c e r e a l g r a s s Bone meal Lime s t o n e I o d i z e d s a l t Manganese s u l p h a t e V i t a m i n A (325,000 I.U./gm) V i t a m i n D3 (33,039.7 I.U./gm) R i b o f l a v i n B a c i f e r m - 25 Amprolium 257. m i x t u r e 7. 29.2 37.8 15 10 3 2 1.5 1 .5 mg/kg .242 .0138 .0165 .0036 .176 4.994 16 Bomb C a l o r i m e t e r . N i t r o g e n was d e t e r m i n e d i n t h e d i e t s and i n t h e e x c r e t a by t h e method o f K j e l d a h l . I n a d d i t i o n t h e f e e d samples were a n a l y z e d f o r t h e i r d r y m a t t e r c o n t e n t . From t h e s e r e s u l t s , the m e t a b o l i z a b l e energy v a l u e s o f t h e d i e t s on a d r y m a t t e r b a s i s and c o r r e c t e d f o r n i t r o g e n r e t e n t i o n , were c a l c u l a t e d as f o l l o w s : M. K c a l p e r gram _ Gross Kcal/gm |~ (Kcal/gm e x c r e t a ) ( g m e x c r e t a ) d r y d i e t d r y d i e t I gm d r y d i e t consumed 8.22 gm N2 r e t a i n e d p e r gm d r y d i e t consumed The n i t r o g e n r e t a i n e d p e r gram d i e t consumed was c a l c u l a t e d by t h e f o l l o w i n g f o r m u l a : Gm N 2 r e t a i n e d / g m d r y B g m N / g m d i e t - (gm ^/gm e x c r e t a ) ( g m e x c r e t a ) d i e t consumed ^ . g m d r y d i e t To d e t e r m i n e the m e t a b o l i z a b l e energy o f t h e t e s t i n g r e d i e n t s on a d r y m a t t e r b a s i s , t h e f o l l o w i n g f o r m u l a was u s e d : M Kcal/gm t e s t i n g r e d i e n t = M Kcal/gm b a s a l d i e t + M Kcal/gm t e s t d i e t - M Kcal/gm b a s a l d i e t grams t e s t i n g r e d i e n t / g m t e s t d i e t The p e r c e n t a g e n i t r o g e n r e t a i n e d was c a l c u l a t e d as f o l l o w s : 17 „ „ . . j gm No i n f e e d consumed - gm No i n e x c r e t a . n n 7o N 2 r e t a i n e d = s & 2 £ x 100 gm N£ i n f e e d consumed I I . Treatment o f samples o f wheat t e s t e d E x p e r i m e n t 1 Wheat No. 1, 13.87. p r o t e i n , u n t r e a t e d , and gro u n d . Wheat No. 1, a u t o c l a v e d f o r 15 m i n u t e s a t 15 l b p r e s s u r e , f a n - d r i e d a t room t e m p e r a t u r e and gro u n d . Wheat No. 1, a u t o c l a v e d f o r 30 m i n u t e s a t 15 l b p r e s s u r e , f a n - d r i e d a t room t e m p e r a t u r e and gro u n d . Wheat No. 1, a u t o c l a v e d f o r 60 m i n u t e s a t 15 l b p r e s s u r e , f a n - d r i e d a t room t e m p e r a t u r e and gro u n d . Wheat No. 2, 12.47. p r o t e i n , u n t r e a t e d , and gro u n d . Wheat No. 2, b o i l e d i n w a t e r f o r 10 m i n . , f a n - d r i e d a t room t e m p e r a t u r e and ground. Wheat No. 2, f r o z e n f o r 3-days a t a p p r o x i m a t e l y -15°C, thawed, and g r o u n d . Wheat No. 2, b o i l e d i n w a t e r f o r 10 m i n . , f r o z e n f o r 3 days a t a p p r o x i m a t e l y -15°C, f a n - d r i e d a t room t e m p e r a t u r e and gro u n d . E x p e r i m e n t 2 Wheat No. 3, 15.47. p r o t e i n , u n t r e a t e d , and ground. Wheat No. 3, a u t o c l a v e d f o r 60 m i n . a t 15 l b p r e s s u r e , f a n -d r i e d a t room t e m p e r a t u r e and gro u n d . Wheat No. 3, soaked i n w a t e r o v e r n i g h t , a u t o c l a v e d f o r 60 min. a t 15 l b p r e s s u r e , f a n - d r i e d a t room t e m p e r a t u r e and gro u n d . 18 Wheat No. 3, soaked i n 1 N HC1 f o r 24 h r , washed t h o r o u g h l y w i t h w a t e r , f a n - d r i e d a t room t e m p e r a t u r e and g'round. Wheat No. 3, soaked i n 1 N HC1 f o r 3 h r , a u t o c a l v e d f o r 60 min. a t 15 l b p r e s s u r e , f a n - d r i e d a t room t e m p e r a t u r e and ground. Wheat No. 3, f r o z e n f o r 5 days a t -4°C, thawed, and ground. E x p e r i m e n t 3 Wheat No. 3, 15.47» p r o t e i n , u n t r e a t e d , and grou n d . Wheat No. 3, a u t o c l a v e d f o r 60 min. a t 15 l b p r e s s u r e , f a n -d r i e d a t room t e m p e r a t u r e and grou n d . Wheat No. 3, soaked i n w a t e r o v e r n i g h t , a u t o c l a v e d f o r 60 min. a t 15 l b p r e s s u r e , f a n - d r i e d a t room t e m p e r a t u r e and grou n d . Wheat No. 3, f r o z e n f o r 5 days a t -4°C, thawed, and ground. Wheat No. 4, 12.17, p r o t e i n , u n t r e a t e d , and grou n d . Wheat No. 4, a u t o c l a v e d f o r 60 min. a t 15 l b p r e s s u r e , f a n -d r i e d a t room t e m p e r a t u r e and grou n d . Wheat No. 4, soaked i n w a t e r o v e r n i g h t , a u t o c l a v e d f o r 60 min. a t 15 l b p r e s s u r e , f a n - d r i e d a t room t e m p e r a t u r e and grou n d . Wheat No. 4, f r o z e n f o r 5 days a t -4°C, and g r o u n d . a f t e r t h a w i n g . E x p e r i m e n t 4 Wheat No. 3, 15.4% p r o t e i n , u n t r e a t e d , and ground. Wheat No. 3, a u t o c l a v e d f o r 60 m i n . a t 15 l b p r e s s u r e , f a n -d r i e d a t room t e m p e r a t u r e , and ground. Wheat No. 3, a u t o c l a v e d f o r 90 m i n . a t 15 l b p r e s s u r e , f a n -d r i e d a t room t e m p e r a t u r e , and ground. 19 Wheat No. 3, a u t o c l a v e d f o r 120 min. a t 15 l b p r e s s u r e , f a n -d r i e d a t room t e m p e r a t u r e , and ground. 20 RESULTS Ex p e r i m e n t 1 The m e t a b o l i z a b l e energy v a l u e s o b t a i n e d f o r t h e samples o f wheat s u b j e c t e d t o d i f f e r e n t t r e a t m e n t s a r e summarized i n T a b l e 2. None o f t h e t r e a t m e n t s imposed had a s i g n i f i c a n t a f f e c t on t h e m e t a b o l i z -a b l e energy v a l u e s o f the samples o f wheat a c c o r d i n g t o t h e a n a l y s i s o f v a r i a n c e ( T a b l e 3 ) . There was a s i g n i f i c a n t d i f f e r e n c e i n t h e p e r c e n t a g e n i t r o g e n r e t e n t i o n from d i e t s c o n t a i n i n g wheat t r e a t e d i n d i f f e r e n t ways ( T a b l e s 2 and 4 ) . A p p l i c a t i o n o f t h e Duncan's m u l t i p l e range t e s t d i d n o t , however, r e v e a l any m e a n i n g f u l d i f f e r e n c e s among th e mean v a l u e s ( T a b l e 5 ) . E x p e r i m e n t 2 The e x p e r i m e n t a l f i n d i n g s a r e p r e s e n t e d i n summarized f o r m i n T a b l e 6. The d a t a show an improvement i n t h e u t i l i z a t i o n o f wheat r e c e i v i n g t h e f o l l o w i n g t r e a t m e n t s : a u t o c l a v i n g f o r 60 min. a t 15 l b p r e s s u r e , w a t e r - s o a k i n g f o l l o w e d by a u t o c l a v i n g f o r 60 m i n . a t 15 l b p r e s s u r e , a c i d - s o a k i n g f o r 3 h r f o l l o w e d by a u t o c l a v i n g f o r 60 min. a t 15 l b p r e s s u r e , and f r e e z i n g f o r 5 days a t -4°C. N e i t h e r a c i d - s o a k i n g f o r 24 h r n o r a c i d - s o a k i n g f o r 24 h r f o l l o x r e d by a u t o c l a v i n g f o r 60 m i n . a t 15 l b p r e s s u r e had any s i g n i f i c a n t e f f e c t on t h e m e t a b o l i z a b l e energy c o n t e n t o f t h e wheat ( T a b l e s 7 and 8 ) . 21 TABLE 2. E f f e c t s o f v a r i o u s t r e a t m e n t s on m e t a b o l i z a b l e energy v a l u e s and % N 2 r e t e n t i o n o f two samples o f wheat i n e x p e r i m e n t 1. No. T r e a t m e n t s 1 Wheat No. 1 U n t r e a t e d ( 1 3 . 8 % p r o t e i n ) M.E. c a l / g d r y m a t t e r 7. N 2 r e t e n t i o n  R e p l i c a t e Average R e p l i c a t e Average 3368.6 3357.0 3365.4 3363.7 50.8 51.2 50.88 50.96 Wheat No. 1 A u t o c l a v e d 15 min. a t 15 l b p r e s s u r e 3372.2 3358.2 3347.8 3359.4 51.13 50.00 50.94 50.69 Wheat No. 1 A u t o c l a v e d 30 min. a t 15 l b p r e s s u r e 3373.0 3361.0 3368.6 3367.5 49.67 52.25 50.56 50.83 Wheat No. 1 A u t o c l a v e d 60 min. a t 15 l b p r e s s u r e 3279.4 3346.6 3420.6 3348.9 49.53 49.01 52.07 50.20 Wheat No. 2 U n t r e a t e d (12.47. p r o t e i n 3342.2 3339.0 3328.2 3336.5 53.08 52.98 51.39 52.48 Wheat No. 2 B o i l e d i n w a t e r f o r 10 m i n . 3339.8 3331.0 3325.4 3332.1 48.74 49.26 50.38 49.46 Wheat No. 2 F r o z e n f o r 3 days a t appro x . -15°C 3411.4 3253.8 3447.8 3371.0 52.64 50.94 52.44 52.01 Wheat No. 2 3309.8 B o i l e d i n w a t e r f o r 10 min. 3331.8 and f r o z e n f o r 3 days a t 3332.6 appro x . -15°C 3324.7 52.82 52.52 51.92 52.42 22 TABLE 3. A n a l y s i s o f v a r i a n c e f o r t h e m e t a b o l i z a b l e energy v a l u e s o f t h e f i r s t e x p e r i m e n t Source o f v a r i a n c e SS d . f . MS F R e p l i c a t i o n 4170.3 2 2085.15 1.04 Treatment 6495.4 7 927.9 .463 E r r o r 2.8048.9 14 2003.5 T o t a l 38714.6 23 TABLE 4. A n a l y s i s o f v a r i a n c e f o r t h e % n i t r o g e n r e t e n t i o n o f t h e f i r s t e x p e r i m e n t Source o f v a r i a n c e SS d . f . MS R e p l i c a t i o n .4428 2 .2214 .229 Treatment 24.6788 7 3.526 3.650* E r r o r 13.5253 14 0.966 T o t a l 39.6496 23 * S i g n i f i c a n t a t 5% l e v e l TABLE 5. R e s u l t o f New M u l t i p l e Range T e s t f o r t h e 7. n i t r o g e n r e t e n t i o n o f t h e f i r s t e x p e r i m e n t T r e a t m e n t s 5 8 7 1 3 2 4 6 Means 52.48 52.42 52.01 50.96 50.83 50.69 50.2 49.46 Mean.; v a l u e s s h a r i n g the same l i n e a r e n o t s i g n i f i c a n t l y d i f f e r e n t a t 57. l e v e l . 24. TABLE 6. E f f e c t o f v a r i o u s t r e a t m e n t s on t h e m e t a b o l i z a b l e energy v a l u e s and 7. ^  r e t e n t i o n o f one sample o f whole wheat i n e x p e r i m e n t 2 No. T r e a t m e n t s 1 Wheat No. 3 U n t r e a t e d (15.4% p r o t e i n ) M.E. c a l / g d r y m a t t e r  R e p l i c a t e Average 3347.3 3323.4 3313.5 3328.1 % N2 r e t e n t i o n  R e p l i c a t e A v erage 47.41 45.84 45.87 46.37 Wheat No. 3 A u t o c l a v e d 60 min. a t 15 l b p r e s s u r e 3399.4 3420.9 3365.6 3395.3* 48.65 45.91 44.92 46.49 Wheat No. 3 3430.0 Water-soaked o v e r n i g h t 3396.5 a u t o c l a v e d 60 min. a t 3456.7 15 l b p r e s s u r e 3427.7* 46.70 45.36 45.68 45.91 Wheat No. 3 A c i d - s o a k e d f o r 24 h r . 3349.6 3360.5 3319.8 3343.3 46.91 46.16 45.89 46.32 Wheat No. 3 3301.8 A c i d - s o a k e d (24 h r . ) and 3329.2 a u t o c l a v e d 60 min. a t 3351.7 15 l b p r e s s u r e 3327.6 44.18 45.65 45.78 45.20 Wheat No. 3 A c i d - s o a k e d f o r 3 h r . a u t o c l a v e d 60 min. 15 l b p r e s s u r e and a t 3365.7 3415.2 3428.6 3403.2* 43.22 46.39 46.75 45.45 Wheat No. .3 F r o z e n a t -4 C f o r 5 days 3368.5 3403.4 3397.4 3389.8* 47.45 49.58 49.04 48.69 * S i g n i f i c a n t l y h i g h e r t h a n t h e u n t r e a t e d sample a t the 57. l e v e l . 25 TABLE 7. A n a l y s i s o f v a r i a n c e f o r t h e m e t a b o l i z a b l e energy v a l u e s o f t h e second e x p e r i m e n t Source o f v a r i a n c e SS d . f . MS F R e p l i c a t i o n 610.6 2 305.3 .4374 Treatment 28941.9 6 4823.65 6.9189** E r r o r 8365.9 12 697.16 T o t a l 37918.4 20 ** S i g n i f i c a n t a t 17. l e v e l TABLE 8. R e s u l t o f New M u l t i p l e Range T e s t f o r t h e M.E. v a l u e s o f second e x p e r i m e n t Treatments 3 6 2 7 4 1 5 Means 3427.7 3403.2 3395.3 3389.8 3343.3 3328.1 3327.6 Mean v a l u e s s h a r i n g t h e same l i n e a r e n o t s i g n i f i c a n t l y d i f f e r e n t a t 57. l e v e l . 26 No d i f f e r e n c e s were a p p a r e n t among t h e v a r i o u s t r e a t m e n t s i n t h e p e r c e n t a g e n i t r o g e n r e t e n t i o n f r o m d i e t s c o n t a i n i n g t h e d i f f e r e n t wheats ( T a b l e 9 ) . Ex p e r i m e n t 3 T a b l e 10 shows t h e e f f e c t s o f v a r i o u s t r e a t m e n t s on t h e m e t a b o l i z a b l e energy v a l u e s o f two d i f f e r e n t samples o f wheat c o n t a i n i n g 15.3% and 12.17. o f p r o t e i n r e s p e c t i v e l y . The n i t r o g e n r e t e n t i o n f r o m t h e d i e t s f o r m u l a t e d w i t h t h e d i f f e r e n t wheats are l i k e w i s e g i v e n i n T a b l e 10. The d a t a from t h i s e x p e r i m e n t were a n a l y s e d u s i n g two-way a n a l y s i s o f v a r i a n c e . The d i f f e r e n c e s i n m e t a b o l i z a b l e energy v a l u e s among t r e a t m e n t s were s t a t i s t i c a l l y s i g n i f i c a n t , T a b l e 11. There was no s i g n i f i c a n t d i f f e r e n c e between t h e m e t a b o l i z a b l e energy v a l u e o f t h e two u n t r e a t e d samples o f wheat. A l s o t h e i n t e r a c t i o n , wheat samples x t r e a t m e n t , was n o t s i g n i f i c a n t . A p p l i c a t i o n o f t h e Duncan's M u l t i p l e Range T e s t i n d i c a t e d t h a t t h e m e t a b o l i z a b l e energy v a l u e s o f t h e two samples o f u n t r e a t e d wheat were n o t s t a t i s t i c a l l y d i f f e r e n t , b u t were s i g n i f i c a n t l y l o w e r t h a n v a l u e s f o r t h e t r e a t e d samples, T a b l e 12. W i t h r e g a r d t o t h e p e r c e n t a g e n i t r o g e n r e t e n t i o n i n t h i s e x p e r i m e n t , t h e a n a l y s i s o f v a r i a n c e ( T a b l e 13) showed s i g n i f i c a n t d i f f e r e n c e s among t h e r e p l i c a t e s , t h e r e f o r e , t h i s t e s t was c o n s i d e r e d to. be i n v a l i d . TABLE 9. A n a l y s i s o f v a r i a n c e f o r t h e p e r c e n t a g e n i t r o g e n r e t e n t i o n o f t h e second e x p e r i m e n t Source of v a r i a n c e SS d . f . MS F R e p l i c a t i o n .664 2 .332 .187 Treatment 23.3542 6 3.89 2.185 E r r o r 21.5980 12 1.78 T o t a l 45.6162 20 28 TABLE 10. E f f e c t o f v a r i o u s t r e a t m e n t s on t h e m e t a b o l i z a b l e energy v a l u e s and 7. N 2 r e t e n t i o n o f two samples o f whole wheat i n e x p e r i m e n t 3 No. T r e a t m e n t s 1 Wheat No. 3 U n t r e a t e d (15.47. p r o t e i n ) M.E. . c a l / g d r y m a t t e r  R e p l i c a t e Average 3342.18 3350.32 3319.80 3337.4 7. No r e t e n t i o n  R e p l i c a t e Average 54.33 55.92 54.63 54.96 Wheat No. 3 3424.24 A u t o c l a v e d f o r 60 min. a t 3391.48 15 l b p r e s s u r e 3408.64 3408.12* 53.61 53.73 55.18 54.17 Wheat No. 3 3419.48 Water-soaked o v e r n i g h t 3406.36 p l u s a u t o c l a v i n g 60 m i n . 3380.40 a t 15 l b p r e s s u r e 3402.08* 55.29 55.45 54.47 55.07 Wheat No. 3 F r o z e n f o r 5 days a t -4°C 3389.64 3419.32 3372.60 3393.85* 55.15 54.60 53.37 54.37 Wheat No. 4 U n t r e a t e d (12.17. p r o t e i n ) 3323.84 3336.64 3302.20 3320.89 54.25 53.43 51.24 52.97 Wheat No. 4 A u t o c l a v e d f o r 60 min. 15 l b p r e s s u r e a t 3402.44 3422.44 3373.40 3399.43* 53.07 55.41 50.09 52.86 Wheat No. 4 3420.80 Water-soaked o v e r n i g h t 3407.36 p l u s a u t o c l a v i n g 60 min. 3382.92 a t 15 l b p r e s s u r e 3403.69* 55.94 54.68 54.25 54.96 Wheat No. 4 F r o z e n f o r 5 days a t -4°C 3393.00 3365.48 3404.52 3387.67* 54.46 55.42 55.13 55.00 * S i g n i f i c a n t l y h i g h e r t h a n the u n t r e a t e d sample a t t h e 57. l e v e l 29 TABLE 11. A n a l y s i s o f v a r i a n c e f o r the m e t a b o l i z a b l e e nergy v a l u e s o f the t h i r d e x p e r i m e n t Source o f v a r i a n c e SS d . f . MS F R e p l i c a t i o n 2229.8 2 1114.9 7.11 Treatment 22680.2 3 7560.06 48.21** E r r o r a 941.2 6 156.8 Wheat 332.5 1 332.5 .847 Treatment x wheat 250.9 3 83.6 .213 E r r o r b 3138.3 8 392.3 T o t a l 29572.9 23 ** S i g n i f i c a n t a t 17. l e v e l TABLE 12. R e s u l t o f new m u l t i p l e range t e s t f o r t h e m e t a b o l i z a b l e energy v a l u e s o f t h e t h i r d e x p e r i m e n t Treatments 2 7 3 6 4 8 1 5 Means 3408.12 3403.69 3402.08 3399.43 3393.85 3387.67 3337.4 3320.9 Mean v a l u e s s h a r i n g t h e same l i n e a r e n o t s i g n i f i c a n t l y d i f f e r e n t a t 57. l e v e l . TABLE 13. A n a l y s i s o f v a r i a n c e f o r t h e 7. n i t r o g e n r e t e n t i o n o f t h e t h i r d e x p e r i m e n t Source o f v a r i a n c e SS d . f . MS F R e p l i e a t i o n 7.1450 2 3.5725 9.623 Treatment 8.3136 3 2.7712 7.4649* E r r o r a 2.2274 6 .37123 Wheat 2.9051 1 2.9051 1.3766 Treatment x wheat 6.2362 3 2.078 .9847 E r r o r b 16.8823 8 2.1102 T o t a l 43.7096 23 * S i g n i f i c a n t a t 5% l e v e l . 31 A p p l i c a t i o n o f Duncan's m u l t i p l e range t e s t d i d n o t r e v e a l any s i g n i f i c a n t d i f f e r e n c e s among t h e mean v a l u e s T a b l e 14. Ex p e r i m e n t 4 The e f f e c t o f a u t o c l a v i n g f o r v a r i o u s p e r i o d s o f t i m e on the m e t a b o l i z a b l e energy c o n t e n t o f wheat and p e r c e n t a g e n i t r o g e n r e t e n t i o n f r o m d i e t s c o n t a i n i n g t h e t r e a t e d wheat i s shown by the d a t a summarized i n T a b l e 15. The sample a u t o c l a v e d f o r 60 min. was s i g n i f i c a n t l y h i g h e r i n m e t a b o l i z a b l e energy c o n t e n t t h a n t h e raw wheat. The d a t a a l s o showed t h a t a u t o c l a v i n g wheat f o r 90 min. and 120 m i n . a t 15 l b p r e s s u r e produced s i g n i f i c a n t l y l o w e r m e t a b o l i z a b l e energy v a l u e s t h a n d i d t h e m i l d e r h e a t t r e a t m e n t (60 m i n . a t 15 l b p r e s s u r e ) and even t h e u n t r e a t e d wheat. The r e s u l t s o f a n a l y s i s o f v a r i a n c e and a p p l i c a t i o n o f Duncan's m u l t i p l e r ange t e s t , a r e p r e s e n t e d i n T a b l e 16 and 17 r e s p e c t i v e l y . V a l u e s f o r t h e p e r c e n t a g e n i t r o g e n r e t e n t i o n f r o m d i e t s c o n t a i n -i n g t h e u n t r e a t e d and t r e a t e d wheat showed t h a t t h e raw and a u t o c l a v e d ( f o r 120 min. a t 15 l b p r e s s u r e ) samples promoted l o w e r n i t r o g e n r e t e n t i o n t h a n t h e samples o f wheat a u t o c l a v e d f o r 60 and 90 m i n . a t 15 l b p r e s s u r e ( T a b l e s 18 and 1 9 ) . F i g u r e 1 r e p r e s e n t s t h e s e f i n d i n g s g r a p h i c a l l y . 32 TABLE 14. R e s u l t o f new m u l t i p l e range t e s t f o r t h e p e r c e n t a g e n i t r o g e n r e t e n t i o n o f t h e t h i r d e x p e r i m e n t T r e a t m e n t s 3 8 7 1 4 2 5 6 Means 5507 55.0 54.96 54.96 54.37 54.17 52.97 52.86 Mean v a l u e s s h a r i n g t h e same l i n e a r e n o t s i g n i f i c a n t l y d i f f e r e n t a t 57. l e v e l . 33 TABLE 15. E f f e c t o f h e a t t r e a t m e n t s on t h e m e t a b o l i z a b l e energy v a l u e s and 7. N 2 r e t e n t i o n o f one sample o f wheat i n e x p e r i m e n t 4 No. T r e a t m e n t s M.E. . c a l / g d r y m a t t e r "LV^ r e t e n t i o n ~~ R e p l i c a t e A v erage R e p l i c a t e Average 1 Wheat No. 4 3371.09 50.57 U n t r e a t e d (15.47. p r o t e i n ) 3379.57 50.64 3359.37 3370.01 51.68 50.96 Wheat No. 4- 3457.17 53.25 A u t o c l a v e d f o r 60 min. a t 3479.21 52.65 15 l b p r e s s u r e 3489.09 3475.16 53.39 5 3 . 0 9 1 Wheat No. 4 3327.37 52.39 A u t o c l a v e d f o r 90 m i n . a t 3296.81 52.82 15 l b p r e s s u r e 3334.01 3319.39 2 52.04 5 2 . 4 2 1 Wheat No. 4 3314.41 48.84 A u t o c l a v e d f o r 120 min. 3322.53 47.58 a t 15 l b p r e s s u r e 3334.25 3323.73 2 48.95 4 8 . 4 6 2 1 S i g n i f i c a n t l y h i g h e r t h a n u n t r e a t e d wheat a t t h e 57. l e v e l . o S i g n i f i c a n t l y l o w e r t h a n u n t r e a t e d wheat a t t h e 57. l e v e l . TABLE 16. A n a l y s i s o f v a r i a n c e f o r t h e m e t a b o l i z a b l e energy v a l u e s o f t h e f o u r t h e x p e r i m e n t 34 Source o f v a r i a n c e SS d . f . MS F R e p l i c a t i o n 311.2 2 155.6 .6598 Treatment 47227.1 3 15742.366 66.75** E r r o r 14.14.9 6 235.82 T o t a l 48953.2 11 * * S i g n i f i c a n t a t 1% l e v e l , TABLE 17. R e s u l t o f new m u l t i p l e range t e s t f o r t h e m e t a b o l i z a b l e energy v a l u e s o f t h e f o u r t h e x p e r i m e n t T r e a t m e n t s Means 3475.16 3370.01 3323.73 3319.39 Mean v a l u e s s h a r i n g t h e same l i n e a r e n o t s i g n i f i c a n t l y d i f f e r e n t a t 57. l e v e l . TABLE 18. A n a l y s i s o f v a r i a n c e f o r t h e 7. n i t r o g e n r e t e n t i o n o f t h e f o u r t h e x p e r i m e n t Source o f v a r i a n c e SS d . f . MS F R e p l i c a t i o n .7072 2 .3536 1.1537 Treatment 37.9652 3 12.6551 41.289** E r r o r 1.8389 6 .3065 T o t a l 40.5113 11 ** S i g n i f i c a n t a t 1% l e v e l . TABLE 19. R e s u l t o f new m u l t i p l e range t e s t f o r t h e 7. n i t r o g e n r e t e n t i o n o f t h e f o u r t h e x p e r i m e n t T r e a t m e n t s Means 53.09 52.42 50.96 48.46 Mean v a l u e s s h a r i n g the same l i n e a r e n o t s i g n i f i c a n t l y d i f f e r e n t a t 57. l e v e l . 3465 r 3435 U 0) "5 £ >_ U) 2 3405 2) _2 3395 o > 0 C ,1* 3365 L -g 3335 N ~0 < 3305 3275 120 T i m e i n m i n u t e s F i g . 1. E f f e c t o f d i f f e r e n t t i m e s o f a u t o c l a v i n g wheat on t h e M. E. v a l u e s and % N£ r e t e n t i o n ( E x p e r i m e n t 4) 37 DISCUSSION I n s i m i l a r s t u d i e s by o t h e r i n v e s t i g a t o r s t h e major c r i t e r i a w h i c h have been used t o e v a l u a t e t h e e f f e c t s o f h e a t t r e a t m e n t on t h e n u t r i t i o n a l v a l u e o f wheat, have been growth r a t e o f t h e e x p e r i m e n t a l a n i m a l s and t h e r a t e and e x t e n t o f p r o t e i n d i g e s t i o n i n v i v o and i n v i t r o . I t i s , however, i m p o r t a n t t o know whether t h e he a t t r e a t m e n t s w h i c h a r e b e n e f i c i a l a c c o r d i n g t o t h e s e c r i t e r i a a l s o a f f e c t m e t a b o l i z -a b l e e n e r g y . A s e r i e s o f e x p e r i m e n t s was co n d u c t e d t o s t u d y t h e e f f e c t o f v a r i o u s t r e a t m e n t s ( a u t o c l a v i n g , f r e e z i n g , w a t e r - s o a k i n g , a c i d - s o a k i n g , b o i l i n g i n w a t e r and c o m b i n a t i o n o f t h e s e t r e a t m e n t s ) on t h e n u t r i t i o n a l v a l u e o f wheat as measured by t h e m e t a b o l i z a b l e energy v a l u e s and by n i t r o g e n r e t e n t i o n f r o m d i e t s c o n t a i n i n g t h e t r e a t e d wheat. I n t h e f i r s t e x p e r i m e n t none o f t h e t r e a t m e n t s w h i c h was used s i g n i f i c a n t l y (P\^.05) improved the energy u t i l i z a t i o n o f t h e two samples o f wheat b u t t h e r e was a s u g g e s t i o n t h a t samples w h i c h had been a u t o c l a v e d f o r 60 m i n . a t 15 l b p r e s s u r e and t h o s e w h i c h had been f r o z e n were s l i g h t l y improved i n m e t a b o l i z a b l e energy v a l u e . The second e x p e r i m e n t was c o n d u c t e d u s i n g t h e same p r o c e d u r e s and r e p e a t i n g t h e s e two t r e a t m e n t s p l u s a c i d and w a t e r s o a k i n g ( T a b l e 6 ) . The e f f e c t o f w a t e r and a c i d - s o a k i n g have been r e p o r t e d by H a s k e l l e t a_l. (1961) and Adams e t al. ( 1 9 6 1 ) , t o improve t h e n u t r i t i o n a l v a l u e o f g r a i n s and g r a i n components. The r e s u l t s o f t h i s e x p e r i m e n t showed t h a t 38 a u t o c l a v i n g f o r 60 min. a t 15 l b p r e s s u r e e i t h e r a l o n e o r a f t e r a c i d -s o a k i n g (3 h r . ) o r w a t e r - s o a k i n g m a r k e d l y i n c r e a s e d t h e m e t a b o l i z a b l e energy v a l u e s o v e r t h e raw wheat. A c i d - s o a k i n g f o r 24 h r . o r a c i d -s o a d i n g (24 h r . ) f o l l o w e d by a u t o c l a v i n g f o r 60 min. a t 15 l b p r e s s u r e d i d n o t a f f e c t m e t a b o l i z a b l e energy as compared t o t h e u n t r e a t e d sample. These l a t t e r f i n d i n g s a r e i n d i s a g r e e m e n t w i t h t h o s e o f Adams ejt a l . (1961) who found t h a t a c i d - s o a k i n g improved t h e n u t r i t i o n a l v a l u e as measured by g r o w t h p r o m o t i n g e f f e c t . T h i s d i s c r e p a n c y may be due t o d i f f e r e n c e s i n t h e c o n c e n t r a t i o n and ti m e o f a c i d - s o a k i n g . The c o n c e n t r a t i o n o f t h e a c i d w h i c h was used by Adams et_ a l . (1961) was n o t s p e c i f i e d . When n i t r o g e n r e t e n t i o n o f c h i c k s f e d d i e t s c o n t a i n i n g the u n t r e a t e d wheat was compared w i t h t h a t o f d i e t s c o n t a i n i n g t r e a t e d wheat samples no d i f f e r e n c e s due t o t r e a t m e n t were o b s e r v e d . From t h e m e t a b o l i z a b l e energy v a l u e s and p e r c e n t a g e n i t r o g e n r e t e n t i o n ( T a b l e 6 ) , i t i s c l e a r t h a t a u t o c l a v i n g , w a t e r - s o a k i n g p l u s a u t o c l a v i n g , and a c i d - s o a k i n g (3 h r . ) f o l l o w e d by a u t o c l a v i n g a f f e c t e d e n ergy u t i l i z a t i o n w i t h o u t a f f e c t i n g n i t r o g e n r e t e n t i o n . T h i s a g rees w i t h Adams e_t a l . (1961) who found t h a t w a t e r t r e a t m e n t a f f e c t e d energy u t i l i z a t i o n , b u t t h e r e was no e f f e c t on p r o t e i n u t i l i z a t i o n as measured by n i t r o g e n r e t e n t i o n . The t h i r d e x p e r i m e n t was c a r r i e d o ut t o compare d i f f e r e n t samples o f wheat g i v e n 3 d i f f e r e n t t r e a t m e n t s ( T a b l e 1 0 ) . The r e s u l t s o f t h i s e x p e r i m e n t c l e a r l y showed t h a t a u t o c l a v i n g f o r 60 min. a t 15 l b p r e s s u r e o r f r e e z i n g f o r 5 days a t -4°C i n c r e a s e d t h e m e t a b o l i z a b l e 39 energy c o n t e n t o f wheat. B o t h samples o f wheat (15.47c. and 12.17o p r o t e i n ) were s i m i l a r l y a f f e c t e d . No s i g n i f i c a n t improvement was found i n t h e p e r c e n t a g e n i t r o g e n r e t e n t i o n f r o m d i e t s c o n t a i n i n g wheat t r e a t e d i n d i f f e r e n t ways ( T a b l e 1 4 ) . T h i s may be due t o t h e r e l a t i v e l y l ow r a t e of- s u b s t i t u t i o n , 257. t e s t i n g r e d i e n t , u s e d . A f o u r t h e x p e r i m e n t was c o n d u c t e d u s i n g t h e same t e c h n i q u e t o d e m o n s t r a t e m a i n l y t h e e f f e c t o f p r o l o n g e d h e a t i n g on t h e m e t a b o l i z a b l e energy v a l u e s o f wheat. The d a t a o f t h i s e x p e r i m e n t , showed marked d e p r e s s i o n i n t h e energy u t i l i z a t i o n when wheat was a u t o c l a v e d f o r 90 m i n . and 120 min. a t 15 l b p r e s s u r e as compared t o u n t r e a t e d wheat. There was a l s o a marked d e c r e a s e i n p e r c e n t a g e n i t r o g e n r e t e n t i o n w i t h t h e d i e t c o n t a i n i n g wheat sample a u t o c l a v e d f o r 120 min. a t 15 l b p r e s s u r e when compared w i t h t h e o t h e r d i e t s c o n t a i n i n g t h e t e s t e d wheat. The a d v e r s e e f f e c t s o f h e a t t r e a t m e n t o f f o o d s t u f f c o n t a i n i n g p r o t e i n and c a r b o h y d r a t e i s w e l l e s t a b l i s h e d and r e v i e w e d (N.R.C. 1950). The r e s u l t s o f t h e f o u r t h e x p e r i m e n t , a l s o , c o n f i r m e d t h e e n h a n c i n g e f f e c t o f moderate h e a t t r e a t m e n t ( a u t o c l a v i n g f o r 60 min. a t 15 l b p r e s s u r e ) on t h e m e t a b o l i z a b l e energy v a l u e s o f wheat ( F i g u r e 1 ) . I t i s w e l l known t h a t i n a d d i t i o n t o soybeans many o t h e r legumes improve i n n u t r i t i v e v a l u e a f t e r t h e a p p l i c a t i o n o f m i l d m o i s t h e a t . I n t h e p r e s e n t work, when wheat was s u b j e c t e d t o t h e c o m p a r a t i v e l y m i l d a u t o c l a v i n g , an a p p a r e n t improvement i n the m e t a b o l i z a b l e energy v a l u e s o v e r t h e u n t r e a t e d wheat, was d e m o n s t r a t e d . T h i s marked i n c r e a s e i n m e t a b o l i z a b l e energy v a l u e s d i d n o t change t o 40 any s i g n i f i c a n t degree when wheat was soaked i n w a t e r o r a c i d b e f o r e a u t o c l a v i n g . The e f f e c t o f a u t o c l a v i n g , p a r t i c u l a r l y on t h e m e t a b o l i z a b l e energy c o n t e n t o f whole wheat, has n o t been r e p o r t e d b e f o r e . However, B e a u d o i n e_t a l . (1951) i n d i c a t e d t h a t c o o k i n g o f whole wheat r e s u l t e d i n a s i g n i f i c a n t improvement i n t h e growth p r o m o t i n g v a l u e f o r t h e r a t . H u t c h i n s o n et^ a l . (1964) u s i n g r a t s a l s o d e m o n s t r a t e d t h a t d i e t s c o n t a i n -i n g f l o u r o r whole meal f r o m t h e steamed wheat promoted a f a s t e r r a t e o f g r o w t h and g r e a t e r f o o d c o n s u m p t i o n t h a n t h o s e c o n t a i n i n g c o r r e s p o n d i n g m a t e r i a l f r o m u n t r e a t e d wheat. The a c t u a l r e a s o n f o r t h i s i n c r e a s e i n n u t r i t i v e v a l u e o f wheat upon h e a t t r e a t m e n t i s n o t known. I n t h e c a s e o f legumes where s i m i l a r phenomena were o b s e r v e d v a r i o u s h y p o t h e s e s have been advanced: r e m o v a l o f a t r y p s i n i n h i b i t o r , e l i m i n a t i o n o f t o x i c s u b s t a n c e s o r d i r e c t a c t i o n on t h e p r o t e i n r e s u l t i n g i n i n c r e a s e d a v a i l a b i l i t y o f c e r t a i n amino a c i d s . H u t c h i n s o n ejt a l . (1964) gave some p o s s i b l e e x p l a n a t i o n s f o r t h i s improvement i n 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 t h e wheat upon h e a t t r e a t m e n t as measured by g r o w t h r a t e and f o o d i n t a k e . They s u g g e s t e d t h a t an improvement i n g r o w t h r a t e may be due t o a r e a l improvement i n t h e n u t r i t i o n a l v a l u e o f t h e p r o t e i n , w h i c h i n d u c e s a f a s t e r r a t e o f g r o w t h and c o n s e q u e n t l y an i n c r e a s e r a t e o f i n t a k e o f f o o d . They a l s o gave c o n s i d e r a t i o n t o t h e p o s s i b i l i t y t h a t a p a r t i c u l a r t r e a t m e n t may have improved t h e p a l a t a b i l i t y o f t h e f o o d so t h a t more i s e a t e n and, 41 when i n t a k e o f p r o t e i n i s t h e f a c t o r l i m i t i n g g r o w t h , t h i s i n c r e a s e d a p p e t i t e promotes a f a s t e r r a t e o f g r o w t h . H u t c h i n s o n e_t a l . d e m o n s t r a t e d a l s o t h a t t h e steam - t r e a t m e n t may have d e s t r o y e d t h e a n t i - t r y p s i n f a c t o r w h i c h i s p r e s e n t i n wheat ( C r e e k e_t a l . , 1961, 1962 and Learmonth and Wood, 1963). The l a s t p o s s i b i l i t y s u g g e s t e d by t h e s e a u t h o r s , and w h i c h t h e y f a v o u r i s t h a t s i n c e t h e t r e a t m e n t d e s t r o y s t h e c a p a c i t y o f t h e wheat p r o t e i n t o f o r m g l u t e n , i t may r e n d e r t h e meal more e a s i l y and e f f i c i e n t l y m a s t i c a t e d and d i g e s t e d . The p o s s i b i l i t y t h a t h e a t i n g may i n c r e a s e t h e p a l a t a b i l i t y and hence i n c r e a s e t h e r a t e o f i n t a k e o f f o o d i s n o t v a l i d i n our e x p e r i m e n t s , s i n c e t h e r e was no n o t i c a b l e d i f f e r e n c e i n f e e d i n t a k e among t h e d i f f e r e n t t r e a t m e n t s . D e s t r o y i n g t h e a n t i - t r y p s i n f a c t o r and/or d e s t r o y i n g t h e c a p a c i t y o f wheat p r o t e i n t o f o r m g l u t e n a r e two p o s s i b l e e x p l a n a t i o n s f o r t h e p r e s e n t f i n d i n g s . A n o t h e r e x p l a n a t i o n i s t h a t h e a t may cause some e s s e n t i a l p r o t e i n f r a c t i o n w h i c h was u n a v a i l a b l e i n t h e raw wheat, t o become a v a i l a b l e f o r a b s o r p t i o n and m e t a b o l i c u s e . T h i s was a l s o s u g g e s t e d by Hayward e t a l . (1936) i n soybean m e a l . The m o i s t h e a t a p p l i e d d u r i n g a u t o c l a v i n g m i ght cause d e n a t u r a t i o n o f some p r o t e i n r e n d e r i n g i t more r e a d i l y d i g e s t i b l e . Thus t h i s may be a n o t h e r p o s s i b l e e x p l a n a t i o n f o r t h e improvement i n n u t r i t i v e v a l u e a c h i e v e d i n t h i s work. 42 The a d v e r s e e f f e c t o f h e a t - t r e a t m e n t on energy u t i l i z a t i o n i s a l s o c o n f i r m e d i n t h e s e s t u d i e s . There was a marked d e c r e a s e i n m e t a b o l i z a b l e energy v a l u e s o f wheat samples a f t e r a u t o c l a v i n g f o r p e r i o d s o f 90 and 120 min. a t 15 l b p r e s s u r e . The impairment o f t h e n u t r i t i v e v a l u e a f t e r t h e s e v e r e h e a t t r e a t m e n t i s a l r e a d y w e l l e s t a b l i s h e d . There a r e many p o s s i b l e t y p e s o f r e a c t i o n s w h i c h may lo w e r t h e b i o l o g i c a l v a l u e o f p r o t e i n . The most i m p o r t a n t ones a r e t h e M a i l l a r d r e a c t i o n s , c r o s s - l i n k a g e r e a c t i o n s , and d i s i n t e g r a t i n g r e a c t i o n s (Donoso e t a l . , 1962). I t was a l s o n o t e d t h a t t h e d e c r e a s e i n t h e m e t a b o l i z a b l e energy v a l u e s upon s e v e r e h e a t t r e a t m e n t was accompanied by a d e c r e a s e i n t h e p e r c e n t a g e n i t r o g e n r e t e n t i o n . I t may be c o n c l u d e d t h a t s e v e r e h e a t t r e a t m e n t s a f f e c t b o t h m e t a b o l i z a b l e energy v a l u e s i n g e n e r a l and p r o t e i n u t i l i z a t i o n i n p a r t i c u l a r . An improvement i n t h e m e t a b o l i z a b l e energy c o n t e n t o f t h e whole wheat has been a c h i e v e d by f r e e z i n g . The mechanism by w h i c h t h e f r e e z i n g p r o c e s s b r i n g s about t h i s improvement i n t h e m e t a b o l i z a b l e energy v a l u e s o f wheat i s unknown. I t appears t h a t t h e change i s i n t h e p h y s i c a l c h a r a c t e r o f some components. Perhaps i c e f o r m i n g i n t h e c o u r s e o f f r e e z i n g caused m e c h a n i c a l s t r e s s , so t h a t a l t e r a t i o n may o c c u r i n some u n a v a i l a b l e f r a c t i o n o f p r o t e i n and as a r e s u l t make the p r o t e i n more r e a d i l y o r c o m p l e t e l y h y d r o l y z e d by d i g e s t i v e enzymes. On t h e o t h e r hand, the change may be a s s o c i a t e d w i t h t h e c a r b o h y d r a t e components. F r e e z i n g c o u l d a f f e c t t h e p h y s i c a l s t r u c t u r e 43 o f t h e c a r b o h y d r a t e t o r e n d e r i t more e a s i l y d i g e s t e d . F u r t h e r e x p e r i m e n t s a r e needed t o e l u c i d a t e t h e s e p o i n t s . 44 SUMMARY A s e r i e s o f e x p e r i m e n t s was c a r r i e d out t o s t u d y t h e e f f e c t s on n u t r i t i v e q u a l i t y o f t r e a t i n g whole wheat i n d i f f e r e n t ways. M e t a b o l i z a b l e energy v a l u e s o f t h e v a r i o u s samples o f wheat were d e t e r m i n e d u s i n g 2 - 3 week o l d c h i c k e n s . N i t r o g e n r e t e n t i o n o f d i e t s c o n t a i n i n g t h e d i f f e r e n t wheat samples, t r e a t e d and u n t r e a t e d were a l s o measured. A u t o c l a v i n g wheat f o r 60 min. a t 15 l b p r e s s u r e , i n c r e a s e d m e t a b o l i z a b l e energy v a l u e s o v e r t h e u n t r e a t e d wheat. F r e e z i n g r e s u l t e d i n s i g n i f i c a n t improvement a l s o . Water- o r a c i d - s o a k i n g d i d n o t improve the m e t a b o l i z a b l e energy v a l u e s o f t h e wheat samples. The v a r i o u s t r e a t m e n t s d i d n o t a f f e c t p r o t e i n u t i l i z a t i o n as measured by p e r c e n t a g e n i t r o g e n r e t e n t i o n f r o m d i e t s c o n t a i n i n g t h e t r e a t e d wheat. P r o l o n g e d h e a t i n g (90 min. and 120 min. a t 15 l b p r e s s u r e ) r e d u c e d t h e energy v a l u e o f whole wheat as w e l l as t h e p e r c e n t a g e n i t r o g e n r e t e n t i o n . V a r i o u s e x p l a n a t i o n s f o r t h e e f f e c t s n o t e d were d i s c u s s e d . 45 REFERENCES Adams, 0. L. and E. C. Naber. 1961. " S t u d i e s on the mechanism o f the c h i c k g r o w t h p r o m o t i n g e f f e c t a c h i e v e d by wa t e r t r e a t m e n t o f g r a i n s and t h e i r components." P o u l t . S c i . 40: 1369-1370. A l l e r d , J . B., L. S. J e n s e n and J . M c G i n n i s . 1957. " F a c t o r s a f f e c t i n g t h e r e s p o n s e o f c h i c k s and p o u l t s t o f e e d p e l l e t i n g . V P o u l t . S c i . 36: 517-523. B e a r s e , G. E., L. R. B e r g , C. F. M c C l a r y and V. L. M i l l e r . 1952. 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