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A survey of the carotene and ascorbic acid content of moose browse Roberts, Leslie Wilson 1948

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A SURVEY OF THE CAROTENE AND ASCORBIC ACID CONTENT OF MOOSE BROWSE  L e s l i e Wilson Roberts  A T h e s i s submitted i n P a r t i a l F u l f i l m e n t of t h e Requirements  f o r t h e Degree o f  Master o f A r t s i n the Department o f Zoology  The U n i v e r s i t y o f B r i t i s h September 1948  Columbia  ABSTRACT The p r o - V i t a m i n A and V i t a m i n C c o n t e n t s o f t h e n i n e main s p e c i e s o f moose browse have been s u r v e y e d . s p e c i e s o f c o n i f e r s have a l s o been surveyed.  Three  Four feeding  a r e a s o f t h e moose, chosen t o r e p r e s e n t a s e r i e s o f s t a g e s i n t h e development o f a f o r e s t f r o m i t s y o u n g e s t t o i t s c l i m a x s t a g e , have been s t u d i e d and compared a s a s o u r c e o f c a r o t e n e and a s c o r b i c a c i d . f o r e s t approximates est content.  A burned over a r e a whose p r e s e n t  twenty y e a r s was found t o have t h e h i g h -  A s e a s o n a l v a r i a t i o n i n v i t a m i n c o n t e n t has  been e s t a b l i s h e d . The d e c i d u o u s  s p e c i e s have a summer max-  imum and a w i n t e r minimum.  The c o n i f e r s have a w i n t e r max-  imum and a summer minimum.  A comparison  of p a l a t a b i l i t y  r a t i n g and v i t a m i n c o n t e n t o f i n d i v i d u a l s p e c i e s i s d i s cussed.  D u r i n g f r e e z i n g and c o l d s t o r a g e t h e l o s s o f c a r o -  tene and V i t a m i n C was found t o be v e r y  slight.  ACMOWLEDGMMTS The author wishes  t o thank Dr. I . MoT. Cowan and D r .  W.S. Hoar f o r I n i t i a t i n g t h i s problem, and t o acknowledge the c o n s t a n t i n t e r e s t , a d v i c e and h e l p so f r e e l y g i v e n t o c a r r y the work t o completion. Thanks a r e a l s o extended  t o Mr. J . H a t t e r who s e l e c t e d  the a r e a s and made the monthly c o l l e c t i o n s o f browse f o r the experiments.  The a s s i s t a n c e o f Mr. J . H a t t e r and Mr.  G. P o t t e r i n the r e p r o d u c t i o n o f the f i g u r e s i s g r e a t l y appreciated. Thanks are owing t o t h e N a t i o n a l Research  Council,  Ottawa, and the P r o v i n c i a l Game Commission, V i c t o r i a , f o r funds t o enable t h i s survey t o be o a r r i e d t o completion.  TABLES AND FIGURES Tables Table I  L i s t o f s p e c i e s by areas  Table II  Carotene v a l u e s f o r unknowns  T a b l e III  A s c o r b i c a c i d v a l u e s f o r unknowns  T a b l e IV  Carotene v a l u e s - Check Area  Table V  Carotene v a l u e s - Area I  T a b l e VI  Carotene v a l u e s - Area II  T a b l e VII  Carotene v a l u e s - Area III  T a b l e VIII A s c o r b i c a c i d v a l u e s - Check Area T a b l e IS  A s c o r b i c a c i d v a l u e s - Area I  Table z  A s c o r b i c a c i d v a l u e s - Area II  T a b l e XL  A s c o r b i c a c i d v a l u e s - Area I I I  T a b l e XII  Carotene l o s s i n f r e e z i n g  T a b l e XIII A s c o r b i c a c i d l o s s i n f r e e z i n g Table XIV  P a l a t a b i l i t y r a t i n g and t o t a l v i t a m i n c o n t e n t  Figures Figure 1  Map o f study a r e a  Figure 2  Check Area  Figure 3  Check Area  Figure 4  Relative positions  Figure 5  Area I  Eigure 6  Area I I  Figure 7  Area I I I  Figure 8  E f f e c t o f storage, carotene  Figure 9  E f f e c t o f storage, a s c o r b i c a c i d  o f Areas I , I I and I I I  TABLES AND FIGURES  (CONT'D)  F i g u r e 10  Carotene c a l i b r a t i o n  curve  F i g u r e 11  Ascorbic  F i g u r e 12  Comparison o f a r e a s ,  carotene  June 5  F i g u r e 13  Comparison o f areas,  carotene  J u n e 29  F i g u r e 14  Comparison o f a r e a s ,  carotene  J u l y 19  F i g u r e 15  Comparison o f areas,  ascorbic acid  June 5  F i g u r e 16  Comparison o f areas,  ascorbic acid  J u n e 29  F i g u r e 17  Comparison o f areas,  ascorbic acid  July  F i g u r e 18  Seasonal v a r i a t i o n s o f deciduous c a r o t e n e , Check A r e a  species  F i g u r e 19  Seasonal v a r i a t i o n s o f deciduous carotene, Area I  species  Figure  20  Seasonal v a r i a t i o n s carotene, Area I I  of deciduous  species  Figure  21  Seasonal v a r i a t i o n s of deciduous oarotene, Area I I I  species  Figure  22  Seasonal v a r i a t i o n s of deciduous a s c o r b i c a c i d , Check A r e a  species  Figure  23  Seasonal v a r i a t i o n s of deciduous ascorbic a c i d , Area I  species  Figure  24  Seasonal v a r i a t i o n s o f deciduous ascorbic a c i d , Area I I  species  Figure  25  Seasonal v a r i a t i o n s o f deciduous ascorbic acid, Area I I I  species  Figure  26  Seasonal v a r i a t i o n s o f c o n i f e r s c a r o t e n e , Check A r e a  Figure  27  Seasonal v a r i a t i o n s of c o n i f e r s a s c o r b i c a c i d , Check A r e a  acid calibration  curve  19  TABLE OF CONTENTS  Introduction  1  Materials and Methods  E  Materials  2  Study Area I  3  Study Area I I  4  Study Area I I I  4  Storage of samples and e f f e c t s  6  Methods  8  Sampling  8  Moisture determination  8  Carotene assay  8  Ascorbic acid determination Discussion  IS 16  Carotene v a r i a t i o n  16  Seasonal v a r i a t i o n  18  Ascorbic acid v a r i a t i o n  EO  Comparison of study areas  23  Vitamin content compared with p a l a t a b i l i t y  25  Summary  26  References  28  Tables  30  Figures  59  INTRODUCTION Numerous studies have been done to determine the v i t amin content of human foods and those of domestic animals. The r e s u l t s of these studies have proved most b e n e f i c i a l to mankind f o r both h i s personal and h i s economic w e l l being. The findings may be applicable to game animals i n cert a i n cases, but the food of the game animals d i f f e r s rather greatly and must be studied to apply any s p e c i f i c f i n d i n g s . This' aspect of game management has been l i m i t e d due to lack of funds and the pressing need f o r other research.  I t i s to  be hoped that the n u t r i t i o n of game animals w i l l be given extensive study i n the near future. The food of the moose, p a r t i c u l a r l y the twigs, consists of various members of the Angiosperms (deciduous species). Three species of Gymnosperms ( c o n i f e r s ) , r a r e l y i f ever eaten by the moose, occurring i n the Check Area have been assayed also,  A survey of the nine main food plants of the moose  (Table I) has been attempted.  This was commenced on the  plants i n v t h e i r winter resting stage (March) and c a r r i e d through u n t i l the summer l e a f i n g i n J u l y . The purpose of t h i s study was to determine: (a) the provitamin A content of the moose browse, (b) the ascorbic aoid browse, (o) the seasonal v a r i a t i o n i n the quantity of the vitamin, (d) a comparison of vitamin content i n d i f f e r e n t feeding areas and (e) a comparison of v i t amin content with p a l a t a b i l i t y .  MATERIALS AND  METHODS  Materials The B.C.  p l a n t s t e s t e d came f r o m  the v i c i n i t y  These were from f o u r d i f f e r e n t  samples were f r o m  the  unpublished Check  and  Areas  Quesnel,  the monthly ones.  The  I , I I and I I I .  o f the a r e a s f o l l o w s (summarized  from  of J . H a t t e r ) .  Area The  Hill" is  data  a r e a s and  same t r e e s o r n e i g h b o u r i n g  a r e a s a r e d e s i g n a t e d as Check Area, A detailed description  of  cheek area i s s i t u a t e d  south o f the Quesnel  an a r e a t y p i c a l  ing area.  on  t h e summit o f "Red  River.  I t was  of t h a t normally used  C o l l e c t i o n s w e r e made a l o n g  Bluff  chosen because i t by moose a s a f e e d -  either  s i d e of the o l d  r o a d between t h e r e d g r a v e l p i t and  the j u n c t i o n w i t h  C a r i b o o Highway.  i n t h i s area i s Douglas  fir  (Pseudotsuga  The  climax f o r e s t  taxifolia).  Due  to the road  m a i n t e n a n c e , many o f t h e t r e e s and and  the ages o f those All  1 and  the s p e c i e s were c o l l e c t e d  from  birch  (Betula glandulosa)  (Salix  sp.) w e r e c o l l e c t e d  (Populus  to  and  collected  on 5).  from  Blackwater Black the  b e a c h o f t h e F r a s e r R i v e r o p p o s i t e t h e mouth o f t h e River.  In  varieties.  Swamp w i l l o w  ( F i g u r e s 1 and  t r i c h o c a r p a ) was  determine.  o r swamp  f r o m a l a r g e bog  Road, 8 m i l e s w e s t o f Q u e s n e l  and  t h i s area (Figures  S) w i t h t h e e x c e p t i o n o f t h e l o w l a n d o r Bog  clearing  have been c l e a r e d  sampled i s d i f f i c u l t  Arctic  poplar  shrubs  the  gravelly Quesnel  t h e c h e c k a r e a t h e n i n e main t y p e s o f browse  and  3. the t h r e e c o n i f e r s s a m p l e d and  (Spruce,  Jaekpine  and  Douglas f i r ) were  assayed.  S t u d y A r e a s I , I I and  I I I are situated  two  miles  north  o f Q u e s n e l a t t h e j u n c t i o n o f t h e C a r i b o o Highway and B a r k e r v i l l e Road. consequently  the  T h e s e t h r e e a r e a s v a r y g r e a t l y i n age  t h e browse i n each a r e a d i f f e r s  and  i n s i z e and  in  t h e amount p r e s e n t . Area  I T h i s area can  ing  operations i n the v i c i n i t y  b e g a n i n 1942, practically the 4  and  the  leveled  The  .2 m i l e s w i d e and  extent  The  sp.)  i n Area  I are l e s s  and  than  30  ( S a l i x sp.)  east  l o i d e s ) a r e a v a i l a b l e , making A r e a  end.  glandu-  trichooarpa)  I I I and midway A l l the  and U p l a n d  be-  trees  the  ex-  willow  T h i s means t h a t  the  (30 i n c h e s ) ,  Aspen (Populus  I a good f e e d i n g  an  exception  birch (Betula  snow l e v e l and  cycle i n  confined to  inches i n height, with tremuloides)  clearing  ( F i g u r e s 1,  south  (Populus  sp.) w h i c h s e l d o m e x c e e d 8 f e e t .  the Upland willow  was  of Area I.  l e s s p a l a t a b l e f o o d s a r e below the but  Bog  to Area  south l i m i t s  c e p t i o n of Aspen (Populus (Salix  sampling  Black'poplar  t a k e n f r o m an a r e a a d j a c e n t  tween t h e n o r t h and  a new  of the c l e a r i n g  the  clear-  slashings  o f browse ( T a b l e I ) w i t h t h e  (Salix  l o s a ) were assayed.  beginning  from  The  of the  .2 m i l e s l o n g a t t h e  seven types  o f t h e Swamp w i l l o w  was  subsequent burning  5) i s a b o u t 2 m i l e s , b u t  I n Area I ,  of the a i r p o r t .  a l l plant l i f e ,  serai formation.  and  area  be d e s c r i b e d a s open, r e s u l t i n g  tremuarea.  4. • Area  II This area  ( F i g u r e s 1,  cession intermediate s o u t h o f , and  ifer  III.  extends south  end  i s a b o u t 20 y e a r s  trees per  acre.  In  Service berry  (Betula p a p y r i f e r a ) , Upland willow  Hazel  (Corylus).  As  s p e c i e s a r e b e l o w 30  inches.  The  Aspen  Paper b i r c h ( B e t u l a p a p y r i f e r a ) are  However, due  old.  order  (Salix  Paper  tremuloides)  tall  sp.)  t o be  as food  b e y o n d optimum f o r  and  deciduous  (Populus too  con-  (Populus  ( S a l i x sp.)  to the c o n i f e r d e n s i t y the  Therefore Area I I i s r e a l l y  The  Aspen  avail-  for  the  snow l e v e l  l o w e r e d m a k i n g more o f t h e l e s s p a l a t a b l e s p e c i e s  is  available.  availability  c o n s i d e r t h e more p a l a t a b l e s p e c i e s , b u t n o t  consider  .25  of density,  i n A r e a I t h e most o f t h e  able, l e a v i n g only Upland willow  we  high-  It is  (Amelanohier f l o r i d a ) ,  birch  when we  ex-  clearing  o f the B a r k e r v i l l e Road.  i n t h i s area  d e n s i t y i s 2163  moose.  It  Cariboo  of the a i r p o r t  the f o l l o w i n g deciduous species are present:  and  suc-  .1 m i l e s w i d e .  forest  tremuloides),  stage i n  I t i s located  j u n c t i o n o f t h e B a r k e r v i l l e and  m i l e s l o n g and The  represents a  upon, t h e B a r k e r v i l l e R o a d .  ways t o t h e e x t r e m e s o u t h e r n where t h i s  6)  b e t w e e n A r e a s I and  bordering  tends from the  4 and  so i f  the l e s s p a l a t a b l e s p e c i e s .  Area I I I This area  ( F i g u r e s 1,  succession nearing m a n n i ) and  4 and  the climax  7)  forest  represents  a  stage  of Spruce ( P i c e a  Balsam f i r (Abies l a s i o c a r p a ) , but  in engel^  containing  5. n o t i c e a b l e amounts of Douglas f i r (Pseudotsuga J a e k p i n e (Pinus o o n t o r t a v a r . l a t i f o l i a ) .  taxifolia)  I t i s bordered  and on  the south by the B a r k e r v i l l e Road, on the e a s t by Area I and on the n o r t h by a road b i s e c t i n g Area I n o r t h o f the l a n d i n g strip;  the west boundary i s formed by the Cariboo Highway.  I t i s approximately 1.7 The o l d e r elements  m i l e s l o n g and  .2 m i l e s wide.  of the f o r e s t of t h i s a r e a a r e over  70 y e a r s of age and t h e r e i s evidence t h a t i t was f o r e s t f i r e about 60 y e a r s ago.  c l e a r e d by  The c o n i f e r d e n s i t y of t h i s  f o r e s t i s approximately 43,620 p e r a c r e .  The c o n i f e r d e n s i t y  being so much l a r g e r than i n the other areas has r e s u l t e d i n the shrubs r e a c h i n g a g r e a t e r h e i g h t .  Consequently most o f  the Aspen (Populus t r e m u l o i d e s) and 35$ of the Upland w i l l o w ( S a l i x sp.) a r e beyond r e a c h o f the moose.  S e r v i c e berry  (Amelanchier f l o r i d a ) i s h i g h e r and thus more i s a v a i l a b l e f o r food.  -Area I I I forms good oover but i s I n f e r i o r t o the  other areas i n q u a n t i t y of a v a i l a b l e f o o d . A comparison  o f Areas I , I I and I I I w i t h r e g a r d t o a v a i l -  a b i l i t y o f Upland w i l l o w ( S a l i x s p . ) , Aspen (Populus tremul o i d e s ) , Paper b i r c h ( B e t u l a p a p y r i f e r a ) and S e r v i c e b e r r y (Amelanchier f l o r i d a ) shows Area I having 4029 i n c h e s , Area 2317  i n c h e s and Area I I I 1178  inches.  U  Thus i n a v a i l a b i l i t y  of food, Area I i s almost twice as g r e a t as Area I I , which i n t u r n i s t w i c e as g r e a t as Area I I I . Sampling  i n the F i e l d  Sampling  c o n s i s t e d of c u t t i n g w i t h a pocket k n i f e the  6.  ends o f branohes,  I n c l u s i v e o f two and t h r e e y e a r o l d wood.  Only one branch was taken from each t r e e o r shrub, and between 12 and 50 i n d i v i d u a l s were sampled collection.  f o r each s p e c i e s  The c l i p p e d ends were t i e d t o g e t h e r i n a bundle  and l a b e l l e d w i t h a number.  Upon completion o f t h e sampling  the numerous bundles f o r eaoh a r e a were p l a c e d t o g e t h e r , wrapped i n damp b u r l a p , l a b e l l e d and shipped by a i r express on t h e day c o l l e c t e d .  I t was n o t p o s s i b l e t o make a complete  c o l l e c t i o n d u r i n g any one day.  Hence, i n most i n s t a n c e s ,  t h e r e i s a two-day spread i n t h e o o i l e d t i n g o f t h e monthly shipments.  D u r i n g h o t weather t h e m a t e r i a l a w a i t i n g t r a n s -  p o r t was kept c o o l and prevented from w i l t i n g by  submerging  the bases o f t h e bundles i n 4 M i l e Creek near Area I . The specimens l e f t Quesnel a t about 9 P.M. and were del i v e r e d t o the U n i v e r s i t y about noon the f o l l o w i n g day.  Only  unbrowsed t r e e s and shrubs were used i n making up t h e monthly collections. Storage On a r r i v a l they were s t o r e d a t 0°F. u n t i l assayed.  This  s t o r a g e temperature was used t o d e l a y o x i d a t i o n o f t h e a s c o r bic aoid or carotene.  A number o f workers r e p o r t t h a t f r e e z -  i n g r e s u l t s i n no l o s s o f c a r o t e n e o r a s c o r b i c a o i d .  Dunker,  F e l l e r s and F i t z g e r a l d (1) found t h a t t h e r e i s no l o s s o f a s c o r b i c a c i d i f c o r n i s f r o z e n and kept a t -10°F.  (-23.3°C).  Fenton and T r e s s l e r (2) found t h a t peas kept a t 0°F. (-17.8°C) and then assayed, showed no a c t u a l l o s s of a s c o r b i c a c i d due  7. to f r e e z i n g .  Any  T r e s s l e r and  King  l o s s was  due  l o s s o f V i t a m i n C,  kept i n a frozen f o u n d no if  t h a t no  condition.  o r no  6 months. small  l o s s up  storage  F i t z g e r a l d and  t_.7.8°C.) and were n o t  As say  i t was  contorta  able  frozen,  not  var.  possible  (7)  Stamberg  a c i d up  (6)  to  a l s o found The  only  commercial  samples a t  temperature.  to obtain  storage, and  The  as r e c e i v e d  0°F. samples into  the  one  small  samples f o r  as-  a two-quart s e a l e r of  Pinus  o f B e t u l a p a p y r i f e r a were  f r e e z i n g compartment o f  the l a b o r a t o r y  refrig-  From t h e s e s e a l e r s samples were taken a t i n t e r v a l s e f f e c t of  f r e e z i n g compartment was degrees higher The  and  storage.  The  temperature i n  the  u s u a l l y 1 4 ° F . ( - 1 0 " C . ) w h i c h was  14  than samples were k e p t a t i n commercial  e f f e c t of  Tables XII f e c t was  ascorbic  the  but were p l a c e d  latifolia  to determine the  age.  that  vegetables  (-20.6"C.) s u f f e r e d  Kraybill  to f r e e z e  them a t  commercial  i n the  erator.  a t -5*F.  (5)  0°F.  from the  placed  store  quick  locker at  frozen condition.  B e a d l e and  (4)  causes  Fellers  (carotene) of  t o 6 months i n most c a s e s .  ( 8 ) u s e d was  Loeffler  that freezing  l o s s o f b o t h c a r o t e n e and  Zsoheile,  Fenton,  further l o s s occurs i f  l o s s of Vitamin A a c t i v i t y  t h a t Rose h i p s f r o z e n  little  a  and  found a l s o  they are kept i n a s o l i d l y  found  (38$).  ( 3 ) w o r k i n g w i t h p e a s , and  working w i t h strawberries, no  to blanching  XIII.  storage The  d e t e r m i n e d was  i s shown I n F i g u r e s  period  of  9 weeks. I n  storage  8 and  stor9  and  f o r w h i c h the  ef-  t h i s p e r i o d l o s s of c a r o t e n e 3n  8. P i n u s o o n t o r t a v a r . l a t i f o l i a was 10.09 fo and. i n B e t u l a p a p y r i f e r a was 10.24$.  The l o s s of a s c o r b i c a c i d i n P i n u s  o o n t o r t a v a r . l a t i f o l i a was 9.22$ and i n B e t u l a p a p y r i f e r a Was 4.71$. Sampling i n the L a b o r a t o r y I n p r e p a r a t i o n f o r a s s a y i n g , t h e samples were taken from commercial storage and immediately out i n t o small s e c t i o n s . I n t h e oase of twigs the o u t t i n g s were 1/2 t o 1 i n c h l o n g . When both l e a v e s and twigs were a v a i l a b l e , t h e l e a f and p e t i o l e and t h e twig t o which i t was j o i n e d were used.  A large  sample was out up o r t o r n by hand i n t o s m a l l p i e c e s , mixed by hand, and then p l a c e d i n quart s e a l e r s and kept i n t h e r e frigerator. for  From these s e a l e r s s u i t a b l e samples were taken  assay.  Moisture  Determinations  S i n c e t h e samples were s t o r e d f o r v a r i a b l e p e r i o d s , and under d i f f e r e n t c o n d i t i o n s , p r i o r t o assay, t h e water was v a r i a b l e and a l l v a l u e s were expressed dry weight o f the m a t e r i a l .  content  i n respect to the  T h i s g i v e s a b a s i s f o r compar-  i s o n whioh l s c o n s t a n t . . To determine the p e r c e n t  moisture,  a 4 gram sample was c a r e f u l l y weighed, p l a c e d i n a s m a l l beaker and d r i e d f o r 48 hours o r more i n a d r y i n g oven a t 95°0.- 100°C. u n t i l a constant weight was reached. Carotene Assay The method used f o r the e x t r a c t i o n of oarotene was t h a t of Moore and E l y ( 9 ) .  I n t h i s method samples a r e c u t i n t o  9. s m a l l p a r t s and p l a c e d I n a Waring B l e n d e r . approximately  150 ml. o f a foaming mixture  h o l by volume and 43$ petroleum  ether).  To t h i s i s added (57$ o f 95$ a l c o -  The blendor i s r u n  f o r 5 minutes, t h e mixture poured i n t o a beaker and t h e supernatant  l i q u i d i s then f i l t e r e d  a separatory f u n n e l .  through g l a s s wool i n t o  To t h e f u n n e l i s added enough water t o  make t h e a l c o h o l c o n c e n t r a t i o n 80$, thus s e p a r a t i n g an a l c o h o l and an ether l a y e r . and  The lower a l c o h o l l a y e r i s drawn o f f  t h e e t h e r l a y e r i s c o l l e c t e d by i t s e l f .  The r e s i d u e and  a l c o h o l l a y e r a r e e x t r a c t e d f o u r times w i t h 25 m l . p o r t i o n s o f ether, and then d i s c a r d e d . tained.  Only t h e ether l a y e r i s r e -  T h i s c o n t a i n s t h e o a r o t e n o i d s and other pigments.  I t i s washed e i g h t o r more times w i t h 100 m l . p o r t i o n s o f water t o remove a l l t r a c e s o f a l c o h o l .  The ether l a y e r i s  now r u n through a d i o a l o i u m phosphate column, whioh adsorbs from t h e a l c o h o l f r e e e t h e r s o l u t i o n a l l pigments except the oarotenoids.  The oarotene  s o l u t i o n i s then made up ( o r r e -  duced, depending on t h e carotene oontent) t o a known volume and read i n a photelometer u s i n g a f i l t e r 420 mu.  Moore suggests  t r a c t be c o n c e n t r a t e d the a d s o r p t i o n column.  transmitting a t  t h a t the ether s o l u t i o n o f t h e ex-  t o 25 - 50 m l . b e f o r e being r u n through T h i s was found unnecessary and con-  s i d e r a b l e ether was r e c o v e r e d .  I n a s s a y i n g twigs t h e s m a l l  amount o f carotene p r e s e n t made c o n c e n t r a t i n g the f i n a l s o lution  necessary.  The oarotene  s o l u t i o n s were compared uC>H a standard s o -  10.  l u t i o n of carotene (Eastman Kodak preparation oonsisting of 90$ B - carotene and 10$ A - carotene).  A standard  curve  was prepared from fresh solutions and used i n a l l comparisons. The Eastman Kodak preparation of oarotenoids was oompared with a potassium dichromate standard.  A petroleum  ether  (Benzin Merok) solution oontaining 5.0 micrograms of oarotene per ml. gives the same photelometer reading as a 0.05$ aqueous solution of potassium dichromate (Analar);  also 0*25  micrograms of carotene gives the same reading as a 0.0025$ solution of potassium dichromate.  This i s i n accordance with  the r e s u l t s found by Barberie, Hoar and Murray ( 1 0 ) . As a oheok on the method, recovery experiments were performed adding known amounts of carotene to one of two ident i c a l samples before blending.  Values of 94$ up to 97$ av-  eraging 96$ recovery i n 6 determinations were obtained. The tswett tubes used f o r the adsorption column were prepared from soft glass test tubes 19 om. long and 2.2 i n diameter,  cm.-  the bottom being blown out and a small piece of  glass tubing attached.  These tubes were h a l f f i l l e d with d i -  calcium phosphate prepared as suggested by Moore and E l y ( 9 ) and a small amount of anhydrous sodium sulfate placed on top of the column to adsorb any water. The actual procedure followed i n this work i s given below: A 4 to 10 gram sample was placed i n a Waring Blendor j a r and 150 ml. of foaming mixture added.  The blendor  was  11. then run f o r f i v e minutes, i f necessary adding alcohol to keep the mixture foaming.  This mixture was poured into a  beaker and the blendor j a r rinsed with a few ml. of foaming mixture.  The mixture was allowed to s e t t l e and the super-  natant l i q u i d poured into a funnel stoppered by a small wad of glass wool.  To the l i q u i d i n the separatory funnel when  thus f i l t e r e d , was added enough water to make the alcohol concentration 80$. The alcohol layer was separated o f f and the residue and alcohol layer extracted four times with 25 ml. portions of petroleum ether.- The combined ether ext r a c t s were then washed with water a t l e a s t eight times using 100 ml. portions of water.  The washed ether solution  was passed through the f i l t e r column being preceded by 30 ml. of petroleum ether and followed by 2 - 30 ml. portions, or u n t i l the ether f i l t r a t e was c o l o r l e s s . This ether solution was read i n a photelometer and the concentration i n >-/ml. read from the c a l i b r a t i o n curve.  In  t h i s survey a Cenoo Sheard Sanford Photelometer was used with a f i l t e r transmitting 420 millimicrons. C a l i b r a t i o n Curve In preparing the c a l i b r a t i o n curve, 5 mgms. of the Eastman Kodak Carotene Preparation were dissolved i n 50 ml. petroleum ether and readings taken of d i l u t i o n s ranging from 8.0  to 0.1 .  This curve i s shown i n Fig.1'0, and Table I I  shows the values from the graph.  IE. Ascorbic Acid In preliminary experiments the method described by B o l i n and Book (11) using 2, 6 - dichlorobenzinoneindophenol f o r a determination was  of both ascorbic and dehydroasoorbic acids  found unsatisfactory due to pigments i n the samples.  Di-  l u t i o n of the extracts to compensate f o r the pigments present gave too low a concentration of ascorbic acid to be measured. The method described by the Association of Agriculture Chemi s t s (12) was  also used but was  found unsatisfactory due  the woody material and the pigments i n the samples. method used i n the determination  to  The  of ascorbic a c i d i s mod-  i f i e d from that of Roe and Oesterling (13).  I t depends upon  the oxidation of ascorbio acid to dehydroasoorbic aoid f o l lowed by ooupling with 2, 4 - dinitrophenylhydrazine  to form  a red ozazone, and with s u l f u r i c acid to produce a red p i g ment.  A comparison of the color produced, with that of  standard ascorbic acid solutions treated i n the same manner i s used as a means of determining the ascorbic acid content. The red color i s measured with a photelometer at a wave length of 530  mu.  I t was proposed to measure ascorbic aoid content by measuring the t o t a l and " f r e e " dehydroascorbio acids, the difference i n these amounts giving the quantity of ascorbio aoid.  This adaption of the Roe and Oesterling. method i s de-  scribed by the Association of Vitamin Chemists (14).  However  due to the pigments present i n the twigs of the samples t h i s  13* method was found unsatisfactory and no satisfactory method was found f o r the determination of "free" dehydroasoorbio acid.  The method was thus used to measure only the t o t a l  (ascorbic plus dehydroasoorbio acids) present i n the sample. A 10 gram sample was used i n a l l cases f o r both twig and twig - l e a f samples.  This was plaoed i n a Waring Blendor j a r  with 100 ml. of 10$ metaphosphorie  acid - 80$ acetic acid so-  l u t i o n , and the blendor run f o r f i v e minutes. mixture was then f i l t e r e d nel  The blended  by suction through a Buchner fun-  and the f i l t r a t e made up to 100 ml. This f i l t r a t e con-  tained the t o t a l (ascorbic and dehydroascorbio) acid i n the sample.  To remove the pigments and change the ascorbic acid  to dehydroascorbio acid the f i l t r a t e was treated with a c i d washed norite, (1 gm. to 50 ml. of f i l t r a t e ) , shaken vigorously, l e f t f o r f i v e minutes, and then f i l t e r e d . trate was then ready f o r assaying.  This f i l -  "Apparently the acetic  aoid i s p r e f e r e n t i a l l y adsorbed on the n o r i t e and active oxygen i s eluted i n quantities s u f f i c i e n t f o r rapid o x i dation (of the ascorbic a c i d ) " (15).  Adsorption of the v i t -  amin i s counteracted by the presence of acetic acid when n o r i t e i s used. 5$ metaphosphorie  A series of d i l u t i o n s was prepared using acid - 10$ acetic aoid with 1$ thiourea  (to s t a b i l i z e the vitamin during subsequent treatment). The d i l u t i o n s ranged from 1/4 to 1/25, depending upon the concentration of the vitamin. For twigs, three d i l u t i o n s of 1/4, 1/7 and 1/10 were used, and f o r leaves and twigs, three  14. d i l u t i o n s o f 1/10,  1/15  and 1/25 were used.  S p e c i a l l y c a l i b r a t e d t e s t tubes ( K l e t t , No.  801) f o r  use i n the Genoo Sheard S a n f o r d Photelometer a r e used.  Into  each of t h r e e t e s t tubes were p i p e t t e d 4 ml. o f the above d i l u t i o n s , two tubes being t e s t e d , one t o s e r v e as a b l a n k . A blank compensates f o r any pigment n o t d e t e c t e d by the eye but d e t e c t a b l e i n the photelometer. t e s t e d was  I n t o each tube to be  p i p e t t e d 1 ml. o f 2$ 2, 4 - d l n i t r o p h e n y l h y d r a z i n e  reagent and the tubes p l a c e d i n a water b a t h a t 37°G. f o r t h r e e hours.  The blank was  set aside.  A t the end o f t h r e e  hours, the tubes were removed from the water and w i t h the blank were p l a o e d i n an i c e b a t h . ml. o f 85$ s u l f u r i c a c i d was  W h i l e i n the i c e bath, 5  added t o eaoh tube, drop by  drop i n n o t l e s s than one minute. was  Finally,  to the b l a n k  added 1 ml. of 2$ 2, 4 - d l n i t r o p h e n y l h y d r a z i n e .  The  tubes were then i n v e r t e d i n the i o e b a t h u s i n g a s m a l l square of wax  paper t o p r o t e c t the f i n g e r s t o p p e r i n g the  tube, shaken g e n t l y , and then p l a o e d i n a r a c k f o r t h i r t y minutes.  The tubes a r e wiped dry and read i n the p h o t e l -  ometer s e t t i n g the instrument t o read 100$ t r a n s m i s s i o n w i t h the blank i n p o s i t i o n .  T h i s w i l l g i v e the c o n c e n t r a -  t i o n p e r ml. and the content of the sample can be oomputed. A l l a s c o r b i c a c i d c o n t e n t v a l u e s were read from a c a l i b r a t i o n curve prepared as f o l l o w s .  The s t o c k standard  was  prepared by d i s s o l v i n g 100 mgms. of 1 - a s c o r b i c a o i d i n 5$ metaphospheric  - 10$ a c e t i o a o i d s o l u t i o n .  T h i s standard  15. contained 1 mgm.  (1000 y/ml.) of ascorbic acid.  The standard  was treated with n o r i t e (1 gm. to 50 ml.), f i l t e r e d , and d i l u t i o n s prepared ranging from .1 >/ml. to 14 >/ml. l u t i o n s were tested i n the same way as the unknowns.  These d i Figure  11 shows the graph of the recorded values and Table I I I the values from the graph. The curve was prepared using: and (c) both n o r i t e and bromine. s u l t s were almost i d e n t i c a l .  (a) n o r i t e , (b) bromine  In a l l three cases the r e -  The stock standard was pre-  pared using 5$ metaphosphorie - 10$ acetic acid solution, but the d i l u t i o n s were made using 5$ metaphosphorie - 10$ acetic acid solution with 1$ thiourea. Addition of thiourea to the stock standard would have prevented the oxidation of ascorbic to dehydroascorbio a c i d .  The presence of thiourea i n the  d i l u t i o n s does not a l t e r the readings from what they would be i f i t were not included.  However since thiourea was used i n  testing the unknowns, i t was used i n preparing the standard curve.  The readings were obtained using a Cenco Sheard San-  ford Photelometer with a green f i l t e r transmitting a t 530 mu.  • 16. DISCUSSION Carotene The in  Variation c a r o t e n e c o n t e n t o f moose b r o w s e i s shown b y a r e a s  Tables I t o IV.  The v a l u e s h a v e b e e n e x p r e s s e d  grams p e r 100 grams f o r b o t h w e t a n d d r y w e i g h t . for  the twig  The v a l u e s  samples a r e u n i f o r m l y low, t h e m a j o r i t y  b e l o w 2 mgms./lOO Check A r e a  as m i l l i -  gms.  However A m e l a n o h i e r f l o r i d a  and Amelanchier  florida,  being i nthe  Cornus s t o l a n i f e r a and  C o r y l u s i n A r e a I I I r e a c h 4.92, 3.66, 3.41 a n d 3.14 mgms./lOO gms. d r y w e i g h t  respectively.  A l o w c a r o t e n e c o n t e n t i n w i n t e r browse i s t o b e exp e c t e d when i t i s c o n s i d e r e d t h a t t h e t w i g s c o n t a i n or no c h l o r o p h y l l , Z s c h e i l e , Beadle  although  t h e y do c o n t a i n o t h e r  and K r a y b i l l  (7) found  chlorophyll t o carotene i s f a i r l y  be d e m o n s t r a t e d .  constant.  I t i s interesting  pigments.  that the r a t i o of  see a r i s e i n c a r o t e n e v a l u e as t h e season will  little  We  should  progresses  t o note  then and this  that the  c o n i f e r s have a v a l u e f o r c a r o t e n e i n t h e w i n t e r  resting  s t a g e a b o u t 1/2 - 1/3 t h a t o f t h e summer v a l u e .  T h i s was  found and  t o be t h e case  f o r Douglas f i r (Pseudotsuga  Spruce ( P i o e a engelmanni).  latifolia) resting  Jackpine  taxifolia)  (Pinus contorta v a r .  h a d , however, 2/3 t h e summer v a l u e i n t h e w i n t e r  stage.  vary g r e a t l y ,  The c h l o r o p h y l l c o n t e n t giving  o f c o n i f e r s does n o t  them t h e f a m i l i a r c o n n o t a t i o n o f " e v e r -  greens". By  J u n e 5 when t h e n e x t  sampling  took p l a c e , t h e s p r i n g  17. s e a s o n was p l e s had  f a i r l y w e l l advanced.  shown a d e f i n i t e i n c r e a s e .  deciduous  s p e c i e s i n a l l a r e a s was  v a r y i n g f r o m a low chier  The m a j o r i t y o f t h e  florida  t o a h i g h o f E7.61  i n Area I I I ) . tene content, the carotene  v a l u e o f 8.68  A l l of the 73$  having  content  the remaining  15.45  ing less  stage.  The  than h a l f  moisture of March The  Of  f l o r i d a did not i n -  times.  In Area  II i t rest-  small l e a v e s w h i c h were  t h e s i z e o f t h e m a t u r e l e a v e s and  their  about twice t h a t of the twig  s a m p l e s o f J u n e 29 almost  represented  samples  the  same a s  of Cornus s t o l a n i f e r a t h e l o w e s t was  t h e browse a t  the  a t the peak o f the growing season. the moisture  the June 5 samples.  c o n t e n t was Of  the  t o be h i g h e r i n v a l u e t h a n  t h e J u n e 5 o r J u l y 19.  The  ap-  samples  those  h i g h e s t c o n t e n t was  ( A r e a I I I ) w i t h 40.18  The  mgms./lOO  of that gms.;  t h a t of B e t u l a g l a n d u l o s a (Check Area) w i t h  mgms./lOO gms.  The  times  the value f o r the winter  June 5 samples had  64$ w e r e f o u n d  p l e was  ten  28.  proximately  10.08  caro-  of the w i n t e r r e s t i n g c o n d i t i o n .  l e a v e s w e r e n e a r m a t u r i t y and  either  gms.  f o r Amelan-  i n c r e a s e o f more t h a n  times over  c o n t e n t was  summer l e v e l ,  assayed  mgms./lOO  increased their  species, only Amelanchier  EI/2  value f o r  f o r Betula p a p y r i f e r a (both  c r e a s e i t s c o n t e n t hy more t h a n f i v e i n c r e a s e d hy  average  mgms./lOO gms.  s a m p l e s had an  The  sam-  The  20.39 mgms./lOO J u l y 19  average  v a l u e o f t h e J u n e 29  sam-  gms.  samples r e p r e s e n t browse a f t e r i t has  passed  18. the peak o f m a t u r i t y . same s i z e  as those  o o n t e n t was a l s o is  beginning  sayed The  The l e a v e s w e r e a p p r o x i m a t e l y t h e  o f t h e J u n e 29 s a m p l e s ,  similar.  The summer s e a s o n  t o wane a b o u t t h i s  time.  o n l y 36$ w e r e h i g h e r t h a n t h o s e  and t h e m o i s t u r e i n the Cariboo  Of t h e samples a s of previous  samplings.  h i g h e s t r e c o r d e d v a l u e f o r J u l y 19 was t h a t o f C o r n u s  stolanifera  ( C h e c k A r e a ) r e a c h i n g 31.66 mgms./lOO gms. a s  compared w i t h a h i g h v a l u e o f 40.18 mgms./lOO gms. by  t h e same s p e c i e s ( A r e a I I I ) i n t h e p r e v i o u s  The  l o w e s t v a l u e was t h a t o f A m e l a n c h i e r  The  average  reached  collection.  florida  (Area I ) .  f o r t h e J u l y 19 s a m p l e s was 20.31 mgms./lOO gms.  T h i s I s almost  i d e n t i c a l w i t h t h e J u n e 29 a v e r a g e  carotene  content. Seasonal The  Variation fact  t h a t 66$ o f / t h e s a m p l e s showed h i g h e r  on J u n e 29 t h a n  on t h e o t h e r d a t e s  sampled, w o u l d l e a d t o  the conclusion that there i s a seasonal v a r i a t i o n . t h i s out, Of  s i x s p e c i e s w e r e compared b y a r e a  To b e a r  ( F i g s . 18 - 2 1 ) .  t h e 24 s p e c i e s , 14 ( 6 6 $ ) show a g e n e r a l t r e n d r i s i n g  M a r c h 28 t o J u n e 29 a n d t h e n d r o p p i n g . 11  values  show a l a r g e d e c r e a s e .  O f t h e s e 14 s p e c i e s ,  F u r t h e r sampling  bear  out this  that  t h e two a r e a s w h i c h a r e m o s t s i m i l a r  (Check Area  seasonal v a r i a t i o n .  would  tend t o  I t i s interesting  to note  ecologically  a n d A r e a I I ) do n o t f o l l o w t h e p a t t e r n a s much  a s do t h e y o u n g a r e a These areas  from  (Area I ) and t h e o l d e r a r e a  ( A r e a s I and I I I ) a r e v e r y d i s s i m i l a r  (Area I I I ) . ecologic-  19. a l l y y e t show s e a s o n a l v a r i a t i o n s v e r y w e l l m a r k e d . d i t i o n s occurring i n Area sunshine  and m o i s t u r e ,  u t i l i z e less Area  I,  may  such as  t h e same m o i s t u r e c o n d i t i o n s .  The  that Area  I , w h i c h i s v e r y open and  years  o l d , i s so  w h i c h the mature f o r e s t i s over suggest  different ror.  content.  Wyns and  content In the  and Area  found  seem t o  I I I needed l e s s  H o g g l e (19)  s o i l was  be due  found  found  an i m p o r t a n t  to er-  sunlight increased  the in  that a higher Nitrogen factor i n producing  Pepkowit?:, L a r s o n ,  G a r d n e r and  a  Owens  t h a t the carotene content i n c r e a s e d w i t h maturity.  Area  I I are very  more m a t u r e .  recently partially w i t h more a v a i l a b l e  s i m i l a r but  The  more s u n l i g h t . a later  Check  have a s l i g h t  difference being  However s i n c e t h e C h e c k A r e a  c l e a r e d f o r a roadway, i t l e f t sunlight.  amount o f s u n s h i n e due  ing  sunshine  c o n d i t i o n s or sampling  i n ages o f t h e v e g e t a t i o n , t h e Check A r e a p r o b a b l y o l d e r and  veg-  60 y e a r s o l d , w o u l d  increased sharply with r i p e n i n g of f r u i t s . and  of  t h a t the younger p l a n t s were r i c h e r  high carotene content. (20)  to  III, in  climatic  A l l a r d y c e and M i l s e n ( 1 8 )  c a r o t e n e c o n t e n t and  similar  has  However t h i s v a r i a b i l i t y may  types of s o i l ,  of  to Area  that the v e g e t a t i o n of Area  t h a t of Area I .  con-  i n t h e more m a t u r e f o r e s t  III with  e t a t i o n o n l y a few  than  the a v a i l a b i l i t y  be c o m p a r a b l e to. t h e a b i l i t y  s u n l i g h t as found  fact  The  T h i s a r e a w h i c h had  to the f o r e s t cover,  T h i s may  result  summer maximum.  the  i s now  i n the v a r i a t i o n  Area  I I appears  has  the been  verges  a lessened exposed  found,  to  giv-  to f o l l o w the  20. s e a s o n a l v a r i a t i o n p a t t e r n q u i t e c l o s e l y , w h i c h may h e d u e to t h e maturing attributed  to differing  Although a  of the vegetation. soil,  the evidence  D i f f e r e n c e s may a l s o h e  c l i m a t e o r sampling  f o r seasonal v a r i a t i o n s  errors. reaching  summer maximum on J u n e 29 i s n o t a b s o l u t e , i t i s s t r o n g l y  suggestive.  I n t h e Check A r e a  o n l y 33 1/3$ o f t h e p l a n t s  show t h e v a r i a t i o n b u t i n A r e a s variation.  I n the four areas  i a t i o n very c l o s e l y . this variation found  62.5$ f o l l o w t h e s e a s o n a l  F u r t h e r sampling would  over a longer p e r i o d .  Other w o r k e r s have  B a r b e r i e and D a v i d s o n  record  thereafter.  similar  Ascorbic Acid The  variation  c o n d i t i o n s i n 1941.  (22) working w i t h w i l d greens  t h a t t h e maximum c o n t e n t steadily  .Hathaway,  a c o n s i s t e n t seasonal  t h i s was c h a n g e d by c l i m a t i c  i s reached  Other  i n July  recent a r t i c l e s  Hoar,  found  and d e c l i n e s ( 2 0 , 21 a n d 23)  findings. Variation  a s c o r b i c a c i d v a l u e s a r e shown b y a r e a s  X X I I - XXV.  var-  demonstrate  seasonal v a r i a t i o n s f o r carotene content.  D a v i s and Keim (21) found but  I , I I a n d I I I 72.2$ show t h e  i n Tables  The J u n e 5 samples a r e a l l q u i t e low, t h e a v -  erage being  54.52 mgms./lOO gms.  stolanifera  ( A r e a I I ) w i t h a v a l u e o f 88.18 mgms./lOO gms.  and  the lowest  i s B e t u l a g l a n d u l o s a (Check Area) w i t h a  v a l u e o f 21.89 mgms./I00 gms. a o i d h a s been found the s p e c i e s .  The h i g h e s t i s C o r n u s  As w i t h carotene, a s c o r b i c  t o vary w i t h the c h l o r o p h y l l content o f  G i r o u d , Ratsimamanga and L e b l o n d  (16) found  El. t h a t t h e a s c o r b i c a c i d was p a r a l l e l present.  T h i s w o u l d mean t h a t a s t h e s p e c i e s bud a n d l e a f ,  the a s c o r b i c a c i d be  true.  The  content r i s e s ,  and t h i s  However n o c o n s t a n t r i s e  ascorbic acid carotene  to the chlorophyll  has been found,  five  and t h e  c o n t e n t w o u l d a p p e a r more v a r i a b l e t h a n t h e  content. v a l u e s f o r t h e s a m p l e s o f J u n e 5 d o show a c o n s i d -  erable increase i n Vitamin C content, ing  has been found t o  t o t e n times  over  the majority increas-  t h e M a r c h E8 s a m p l e s .  The a v e r a g e  v a l u e f o r J u n e 5 i s 317.33 mgms./lOO gms. w i t h a h i g h f o r Populus  tremuloides  value  ( A r e a I ) o f 5S6.53 mgms./lOO gms* a n d  a l o w v a l u e o f 65.05 mgms./lOO gms. f o r A m e l a n c h i e r  florida  (Check A r e a ) . The  a s c o r b i c a c i d v a l u e s f o r b o t h J u n e E9 a n d J u l y 19  show c o n s i d e r a b l e v a r i a t i o n . seems t o h a v e b e e n r e a c h e d  I n some c a s e s  value  i n t h e J u n e E9 sample, a n d i n  others i t c o n t i n u e s t o r i s e u n t i l J u l y 19. i t was f o u n d  t h e maximum  In still  t o h a v e i n c r e a s e d w i t h J u n e 5 sample,  others  decreased  w i t h t h e J u n e E9 sample a n d t h e n i n c r e a s e d a g a i n w i t h t h e J u l y 19 s a m p l e .  I n t h e J u n e E9 sample two s p e c i e s r e a c h e d  h i g h v a l u e o f o v e r 730 mgms./lOO gms. w h i l e t h e l o w e s t  a  value  r e c o r d e d was f o r C o r y l u s ( C h e c k A r e a ) w i t h a v a l u e o f E14.38 mgms./lOO gms.  The h i g h e s t v a l u e s o b t a i n e d f o r t h e J u l y 19  sample w e r e a b o v e t h o s e o f J u n e 29. corded 100  from Area  I I , being Populus  gms.) a n d P o p u l u s  These were b o t h r e -  tremuloides  (8S4.98 mgms./  t r i c h o c a r p a (855.38 mgms./100. gms.).  The  average  v a l u e f o r J u n e E9 i s 41E.61 mgms./lOO gms. a n d  f o r J u l y 19 i s 471.77 mgms./lOO gms. The  seasonal v a r i a t i o n found  f o rthe carotene  does n o t appear f o r t h e a s c o r b i c a c i d . common t o a l l f o u r a r e a s ,  content  Of t h e £4 s p e c i e s  o n l y 37 1/E $ r e a c h t h e summer  maximum o n J u n e E9, b u t 54.16$ r e a c h t h e summer maximum o n J u l y 19.  Sampling  understanding  beyond t h i s p o i n t would p e r m i t  of the seasonal v a r i a t i o n s of ascorbic a c i d .  F i g u r e s EE - S5 show t h e v a r i a t i o n s a s r e c o r d e d . seen  a clearer  t h a t t h e Check Area,  Areas  I twill  be  I a n d I I h a v e t h e summer  maximum o n J u l y 19 i n m o s t c a s e s . s p e c i e s agree w i t h t h i s f i n d i n g .  I n Area I , f i v e  of s i x  A r e a I I I however f o l l o w s  more t h e p a t t e r n o f t h e c a r o t e n e v a r i a t i o n ,  having  four of  t h e s i x s p e c i e s w i t h t h e summer maximum on J u n e E 9 . The  c o n i f e r s were assayed  the f o u r sampling  dates,  conditions of light, been n o t e d  o n l y i n t h e Check A r e a f o r  and were thus  soil  and c l i m a t i c  subject to similar conditions.  before, the i n c r e a s e o f Carotene  As has  values i s very  o  small i n the conifers. ure  T h i s i s shown i n F i g u r e E6.  E7 we s e e t h a t t h e s e a s o n a l v a r i a t i o n f o r t h e c o n i f e r s  s e t s a p a t t e r n o f i t s own. before the onset  a t i t s lowest,  T h e M a r c h E8 s a m p l i n g  occurred  o f s p r i n g weather and a t t h a t time  c o n d i t i o n s were i n f o r c e . is  In Fig-  We f i n d  that the carotene  but the ascorbic acid  content  content i s a t i t s  h i g h e s t and d e c r e a s i n g r a p i d l y a s t h e t e m p e r a t u r e the spring c o n d i t i o n the a s c o r b i c a c i d  winter  rises.  In  content i s s t i l l de-  23. creasing, reaching a minimum on June 29 which i s close to the peak of the summer season ( f o r deciduous species). Then the value begins to increase again as the temperature approaches the winter conditions.  This v a r i a t i o n was found  also by Griaznov and Alexleeva (17) using b i o l o g i c a l assaymethods. One of the purposes of the study was to compare the carotene and ascorbio acid  v  irt. the four feeding areas. The  comparison i s set out i n Figures 12, 13 and 14 f o r carotene. The same species i s compared i n the four areas f o r each c o l l e c t i o n date. In the c o l l e c t i o n of. March 28 the values were too low to be comparably set out, so t h i s date was not used.  On the  c o l l e c t i o n date of June 5, the Check Area and Area I I I are well above the average, with Areas I and I I either average or below the average i n the majority of species.  Of the 7  species assayed the order of "high" values ( i . e . maximum f o r the p a r t i c u l a r species on the sampling date) i s : Area I I I - 4, Check Area - 2, Area I I - 1, and Area I - 0.  On the June 29  c o l l e c t i o n date the order of "high" values i s : Area I I I - 4, Area I I - 2, Area I - 1, and Check Area - 0.  On the J u l y 19  c o l l e c t i o n date the order i s : Cheek Area - 4, Area. I I - 2, Area I I I - 1 and Area I - 0.  The order f o r the three dates  would then be Area I I I - 9, Check Area - 6, Area I I - 5, and Area I - 1.  This would mean that Area I I I , which i s a mature  forest, has a higher pro-vitamin^content than the other areas,  24. with  the  two  next  highest i n content.  mature a r e a tent.  Using  s i m i l a r areas  ecologically, only the  Area  I - 0.  Area has  is  three areas.  II)  being  the lowest p r o - v i t a m i n A  Area  concluded  of carotene  oon-  that Area  III i s a  t o t h e moose a s i n  P a r t i c u l a r l y when we  the  consider that I than  b e t t e r f e e d i n g area as regards p r o - v i t a m i n A  The  comparison of the a s c o r b i c a c i d  f o r the c o l l e c t i o n  dates  are  food  i n Area  content  of the  shown i n F i g u r e s  vary g r e a t l y from the order f o r carotene  15-17. species  I I - 8 1/2,  1-8,  This order  pletely  reverses Area  f o r carotene tent. is  Area  Area  I I I a s a good f e e d i n g a r e a a s  v a l u e s , and I I , w h i o h was  places i t l a s t the  the best f o r a s c o r b i c a c i d ,  by A r e a  I I I - 0.  com-  found  being followed very  con-  carotene closely  I.  A compilation of "high values" f o r both oarotene ascorbic  Area  i n ascorbic aoid  second b e s t a r e a f o r  On  dates,  the values are i n the f o l l o w i n g order: Area and  four  content.  t h e computed " h i g h " v a l u e s f o r t h e t h r e e c o l l e c t i o n  C h e c k A r e a - 1 1/2,  a  content.  areas ranged i n order of " h i g h " v a l u e s f o r the  assayed  - 4  the food i n t h a t  I I I d o e s i t a p p e a r t h a t t h e more m a t u r e s t a n d w o u l d b e  areas  the  Check A r e a  a v a i l a b l e i n f a r greater quantity i n Area  The  im-  I I - 5,  although  area i s not as r e a d i l y a v a i l a b l e other  Area  I, which i s a r e l a t i v e l y  I I I - 9,  I t must be  much b e t t e r s o u r c e  and  6 s p e c i e s common t o t h e f o u r a r e a s  t r u e o r d e r becomes: A r e a and  (Cheek A r e a  a c i d g i v e s the order:  Area  I I - 14,  Area  and  III -  9,  25. Area 1 - 7 inferred  1/S a n d Check A r e a - 5 1/2.  that a d i s t r i c t  approximately  20 y e a r s  From t h i s i t c a n be  such a s A r e a I I where t h e f o r e s t i s  old, high  enough t o r e d u c e t h e snow  l e v e l , w o u l d b e most a d v a n t a g e o u s a s a f e e d i n g a r e a . o l d e r mature f o r e s t  (60 y e a r s  valuable feeding area, cleared area.  o l d ) w o u l d be t h e s e c o n d  The C h e c k A r e a ( a r e a r e c e n t l y b r o w s e d b y t h e valuable feeding area  i n respect  content.  A comparison o f t h e t e n t a t i v e p a l a t a b i l i t y tablished  by J . H a t t e r ) w i t h  corbic  o f a s p e c i e s f r o m one a r e a  t h i s comparison a o i d found  d a t e s was t o t a l l e d study  areas  o fthe  due t o t h e v a r i a b i l i t y be-  tween s p e c i e s o f t h e same p a l a t a b i l i t y r a t i n g , variability  rating (es-  the vitamin content  moose b r o w s e i s v e r y d i f f i c u l t ,  termine  most  f o l l o w e d c l o s e l y b y t h e open r e c e n t l y  moose) w o u l d be t h e l e a s t to vitamin  The  the content  and a l s o t h e  t o another.  o f carotene  To d e -  and o f a s -  f o r e a c h s p e c i e s on t h e f o u r  sampling  f o r ( a ) t h e Check A r e a and (b) t h e f o u r  (Table XIV).  The c a r o t e n e  content  o f t h e No. 1  palatable  s p e c i e s i s h i g h e s t i n t h e Cheek A r e a , w i t h t h e  exception  of Salix  rating.  sp. (upland willow)  The N o . 4 p a l a t a b l e s p e c i e s i s s e c o n d h i g h e s t i n  t h i s area,  f o l l o w e d b y t h e two N o . 3 ' s .  s p e c i e s has t h e lowest  t o t a l content.  t h e s i x s p e c i e s common t o a l l a r e a s found  whioh has a very low  The N o . 2 p a l a t a b l e  When t h e t o t a l s o f  are considered  i t i s  t h a t t h e N o . 1 p a l a t a b l e s p e c i e s have t h e h i g h e s t  carotene  content,  f o l l o w e d b y t h e No. 4 a n d t h e n  t h e No. 3  26. rated  speoies. Ascorbic  aoid  t o t a l s do n o t show a . t r e n d  that of the oarotene values.  I n t h e C h e c k A r e a one o f t h e  most p a l a t a b l e s p e c i e s h a s t h e h i g h e s t other  value  but the three  s p e c i e s r a t e d most p a l a t a b l e h a v e t h e t h r e e  ascorbic ever,  acid values.  the order  atability  i s highest,  Apart  of total  rating.  four areas  from these  content  show no d e f i n i t e followed  by a No.  palatability  this  rating  lowest  s p e c i e s , how-  c l o s e l y follows the p a l -  trend.  i n d i c a t e s that there i s l i t t l e present  three  The a s c o r b i c a c i d c o n t e n t A Ho.  1, w i t h  A consideration of both carotene  in  similar to  t o t a l s f o r the  3 palatable  t h e No.  speoies  4 species  and a s c o r b i c a c i d  c o r r e l a t i o n between  next.  totals the  and t h e v i t a m i n c o n t e n t  as found  survey.  SUMMARY The n i n e m a i n  s p e c i e s o f moose b r o w s e h a v e b e e n  veyed t o determine t h e i r carotene  and a s c o r b i c a c i d  Three s p e c i e s o f c o n i f e r s found i n a s s o c i a t i o n w i t h moose browse h a v e a l s o b e e n Four areas  a source  a r e a was  representing different  stages  source was  o f o a r o t e n e and a s c o r b i c a c i d .  found  t o be a good s o u r c e  of ascorbic acid.  a poor  source  content. the  surveyed. i n the f o r e s t  s u c c e s s i o n were s t u d i e d t o determine t h e i r r e l a t i v e as  sur-  The most  of carotene  merits mature  but a poor  The a r e a most r e c e n t l y c l e a r e d  of carotene  b u t a good s o u r c e  of ascorbic  27. aoid.  The area midway i n suecession between these two areas  was found to be a valuable source f o r both carotene and ascorbic acid.  The fourth area studied was similar to the  general moose area but had been used as a feeding area only recently.  This area was found to be a poor source of both  por-vitamin A and Vitamin C. The browse studied was found to have a seasonal vari a t i o n increasing up to a summer maximum and then declining. The conifers were found to reverse t h i s v a r i a t i o n , having the maximum content under winter conditions and decreasing to a summer minimum. The vitamin content of moose browse has been compared* with the suggested p a l a t a b i l i t y rating but l i t t l e c o r r e l a t i o n has been found.  28. REFERENCES f  1.  D u n k e r , C.F., F e l l e r s ,  C.R. a n d F i t z g e r a l d ,  CA.,  F o o d R e s . , 4:238-246, 1 9 3 9 . 2.  F e n t o n , F . , a n d T r e s s l e r , D.K., F o o d R e s . , 3:409-416, 1938.  3.  F e n t o n , F . , T r e s s l e r , D.K., a n d K i n g ,  C.G.,  J • N u t r . , 12:285, 1 9 3 6 . 4.  L o e f f l e r , H . J . , J . R e s . , 11:69-83, 1946.  5.  Fitzgerald,  G.A.,  and F e l l e r s ,  C.R.,  F o o d R e s . , 3:109-120, 1 9 3 8 . 6.  Stamberg,  O l o f E . , F o o d Res., 10:392-395, 1 9 4 5 .  7.  _ s e h e i l e , F.P., B e a d l e , B.W.,  and K r a y b i l l ,  H.R.,  F o o d R e s . , 8:299-313, 1 9 4 3 . 8.  Vanoouver  I c e and C o l d S t o r a g e , V a n c o u v e r , B.C.  9.  Mfoore, L.A., a n d Roy E l y , I n d . a n d E n g . Chem. A n a l . E d . , 13:600-601, 1 9 4 1 .  10.  B a r b e r i e , M.A.,  H o a r , W.S.,  a n d M u r r a y , M.J.,  A c a d i a n N a t u r a l i s t , v o l . 2, n o . 5, p . 28-33, 1 9 4 7 . 11.  B o l i n , D.W.,  and L u c i l l e  Book,  S c i e n c e , K o v . 7, v o l . 106, p . 451, 1 9 4 7 . 12.  J o u r n a l o f A s s o c i a t i o n o f O f f i c i a l A g r . Chemists, 1945.  13.  R o e , J . H . , and O e s t e r l i n g , M.J., J. Biol.  14.  Chem., 152:511-517, 1 9 4 4 .  Methods o f V i t a m i n Assay, A s s o c i a t i o n  o f Vitamin Chemists,  I n t e r s c i e n c e P u b l i s h e r s I n c . , New Y o r k , 1 9 4 7 . 15.  Roe,. J . H . , a n d K u e t h e r , C.A., J. Biol.  Chem., 147:399, 1 9 4 3 .  29. REFERENCES (CONT'D) 16.  Giroud, A., Ratsimamanga, R., and Leblond, CP., C.R. Soc. B i o l . 117:612-614,  Nutrition  Abstracts and Review, 4:776, 1934/35. 17.  Grlaznov, N.I., and Alexieeva, E.N., Problems of N u t r i t i o n Moscow, 3, no.5, 57-61, 1934. N u t r i t i o n Abstracts and Reviews, 4:778, 1934/35.  18.  Allardyce, J . , and Milson, D., Canad. J . Res., (C), 20:85-88, 1942.  19.  Wynd, F.L., and Noggle, G.R., Food Res., 11:148-158,.1946.  20.  Pepkowitz, L.P., Larson, R.E., Gardner, J . , and Owens, G., Proc. Amer. Soc. Hart. S c i . , 44:468, 1944.  21.  Hathaway, I.L., Davis, H.P., and F.D. Keim, Univ. Nebraska C o l l . A g r i c , Agric. Exp. Stat. Res. B u l l . , No. 140, Oct. 1945, p. 15.  22.  Hoar, W.S.,  Marjorie Barberie, and Davidson,  W.S.,  Journal of Canadian D i e t e t i c Assn., June, 1948. 23.  Stillman, J.T., Watts, B.M., and Morgan, A.F., J . Home Econ., 35:28-34, 1944.  30. TABLE I SPEGIBS ASSAYED BY AREAS Check Area  Area I  Betula papyrifera ("Paper birch)  Betula papyrifera (Paper hiroh)  Amelanchier f l o r i d a IService berry)  Amelanchier f l o r i d a I Service berry)  Cornus s t o l a n i f e r a HfRed Osier dogwood)  Cornus s t o l a n i f e r a CRed Osier dogwo od)  S a l i x sp. (Upland willow)  S a l i x sp. (Upland willow)  Betula glandulosa ("Bog hiroh)  Populus tremuloides (Aspen)  S a l i x sp. (Swamp willow)  Populus trichooarpa ("Slack poplar)  Populus tremuloides (Aspen)  Corylus (Hazel)  Populus trichooarpa CSlaek poplar) Corylus ^Hazel) Pinus contorta var. l a t i f o l i a (Jackpine) Pseudotsuga t a x i f o l i a IDouglas f i r ) Picea engelmanni (Spruce)  31 TABLE I (CONT'D) SPECIES ASSAYED BY AREAS Area I I I  Area I I Betula papyrifera ("Paper birch)  Betula papyrifera [Paper birch)  Amelanohier f l o r i d a (Servioe berry)  Amelanchier f l o r i d a (Service berry)  Cornus s t o l a n i f e r a CRed Osier dogwood)  Cornus s t o l a n i f e r a (Red Osier dogwood)  S a l i x sp. (Upland willow)  S a l i x sp. (Upland willow)  Populus trenruloides ( Aspen)  Populus tremuloides (Aspen)  Corylus (Hazel)  Corylus  (Hazel)  TABLE I I CAROTENE VALUES FOR UNKNOWNS Reading  r/ml.  Reading  B5 26 27 28 29  2.48 2,41 2.34 2.28 2.21  30 31 32 33 34 35 36 37 38 39  2.15 2.09 2.03 1.97 1.92 1.86 1.81 1.76 1.72 1.67  60 61 62 63 64 65 66 67 68 69  40 41 42 43 . 44 45 46 47 48 49  1.62 1.58 1.53 1.49 1.45 1.41 1.37 1.33 1.29 1.25  50 51 52 53 54 55 56 57 58 59  1.21 1.18 1.15 1.11 1.08 1.04 1.01 0.98 0.95 0.92  ,  y/mi.  0.89 0.86 0.83 0.80 0.78 0.74 0.71 0.69 0.65 0.63  70 71 72 73 74 75 76 77 78 79  0.60 0.58 0.55 0.52 0.50 0.48 0.45 0.42 0.40 0.38  80 81 82 83 84 85 86 87 88 89  0.36 0.34 0.31 0.29 0.27 0.25 0.23 0.20 0.18 0.16  90 91 92 93 94 95  0.14 0.12 0.10 0.09 0.07 0.05  TABLE I I I ASCORBIC ACID VALUES FOR UNKNOWNS Reading  y/mi.  Reading  jr/ml.  39  13.60  69  5; 22  40 41 42 43 44 45 46 47 48 49  13. SO 12.80 12.50 12.12 11.80 11.50 11.15 10.87 10.51 10.25  70 71 72 73 74 75 76 77 78 79  5.03 4.83 4.62 4.40 4.20 4.00 3.80 3.61 3.41 3.22  50 51 52 53 54 55 56 57 58 59  9.98 9.70 9.40 9.13 8.85 8.60 8.32 8.08 7.80 7.55  80 81 82 83 84 85 86 87 88 89  3.05 2.88 2.68 2; 50 2.32 2.15 1.99 1.80 1.60 1.43  60 61 68 63 64 65 66 67 68  7.30 7.05 6.80 6.60 6.38 6.13 5.90 5.65 5.50  90 91 92 93 94 95 96 97 98  1.30 1.18 0.99 0;80 0.63 0.50 0.33. 0.19 0.00  TABLE IV CHECK AREA CAROTENE IN SAMPLES Species Betula papyrifera (Paper birch)  Amelanohier f l o r i d a (Service berry)  Cornus s t o l a n i f e r a (Red Osier dogwood)  Palatabili t y Rating Date 1 -  Weeks i n Sample Percent Storage No. Moisture  Mgms. per100 gms Dry Wet  Mar.28  9  9  36.84  0.73  1.16  June 5  4  9  74.72  4.44  17.56  June 29  3  9  73.51  7.81  29.48  J u l y 19  2  9  73.45  4.69  17.66  Mar.28  8  3  27.85  3.55  4.92  June 5  2  3  74.90  5.04  20.08  June 29  3  3  59,56  8.75  21.64  J u l y 19  2  3  66.54  9.70  28.99  Mar. 28  8  1  26.22  0.26  0.35  June 5  2  1 .  72.87  4.93  18.17  June 29  3  1  70.62  8.75  29.78  July 19  2  1  68.26  10.05  31.66  •  1  1  TABLE I V (CONT'D) CHECK AREA CAROTENE IN SAMPLES Speoies S a l i x sp. (Upland w i l l o w )  Betula glandulosa (Bog b i r c h )  S a l i x sp. (Swamp w i l l o w )  Paiatabili t y Rating IA  2  2  Date  Weeks i n Sample Peroent Storage No. M o i s t u r e  Mar. 28  9  June 5  2  June 29  7  Mgms. p e r 100 gms Wet Dry.  42.87  0.77  1.34  7  72.88  3.45  12.72  3  7  42.77  6.58  11.50  J u l y 19  2  7  67.24  8.97  27.38  Mar. 28  9  16  20.02  0.83  1.04  June 5  3  16  74.14  2.93  11.33  June 29  3  16  66.87  3.34  10.08  J u l y 19  2  16  68.33  3.25  10.26  Mar. 28  9  12  41.82  0.41  0.701  June 5  2  12  78.48  3.09  14.36  June 29  3  12  66.86  4.46  13.46  J u l y 19  2  12  66.07  7.03  20.72  TABLE I V (CONT'D) CHECK AREA CAROTENE IN SAMPLES Species Populus tremuloides (Aspen)  Populus t r i e h o o a r p a (Black poplar)  Corylus (Hazel)  Palatabili t y Rating 3  3  4  .  Date  Weeks i n Sample P e r c e n t Storage No. M o i s t u r e  Mgms. p e r 100 gms Wet Dry  Mar. 28  8  4  41.94  0.54  0.93  June 5  2  4  72.31  5.90  21.31  June 29  3  4  52.09  10.13  17.49  J u l y 19  2  4  55.64  10.13  22.84  Mar. 28  9  15  0.89  1.49  June 5  4  15  40.43 71.43  4.99  17.46  June 29  3  15  61.60  6.88  17.91  J u l y 19  2  15  72.77  5.97  21.92  Mar. 28  9  5  34.54  0.40  0.61  June 5  2  5  71.19  4.13  14.34  June 29  3  5  64.49  8.70  24.50  J u l y 19  2  5  65.28  8.35  24.05  TABLE 17 (CONT'D) CHECK AREA CAROTENE IN SAMPLES Speoies  Palatabili t y Rating  P i n u s o o n t o r t a var. l a t i f o l i a [Tackplne)  Pseudotsuga t a x l f o l i a (Douglas f i r )  P i c e a engelmanni  5  5  6  Date  Weeks i n Sample P e r c e n t No. M o i s t u r e Storage  Mar. 28  9  June 5  2  June 29  3  J u l y 19  2  Mar. 28  8  June 5  2  June 29  5  J u l y 19  2  Mar. 28  9  June 5  1  June 29  3  J u l y 19  2  TSpruceJ  Mgms. p e r 100 gms. Bry Wet-  18  49.32  3.44  6.78  18  59.99  3.03  7.57  18  64.73  3.49  9.90  18  63.29  2.69  7.33  2  48.53  1.33  2.58  2  59.88  1.95  4.86  2  41.90  4.48  7.71  a  59.64  2.50  6.19  17  17.74  2.53  3.08  17  60.81  2.60  6.63  *  17  64.58  2.72  7.68  17  63.29  2.69  7.33  TABLE V AREA I CAROTENE IN SAMPLES Speoies Betula papyrifera (PaperbirchJ  Amelanchier f l o r i d a (Service berry)  Cornus s t o l a n i f e r a fR~ed Osier dogwood)  Weeks i n Sample Percent Storage No. Moisture  Palatabil-  1  s. per 100 gms. Wet Dry  Mar. 28  9  32  27.28  0.55  0.76  June 5  3  32  74.73  4.48  17.73  June 29  4  32  59.82  5.86  14.58  J u l y 19  2  32  74.64  4.82  19.01  Mar, 28  9  34  38.57  0.25  0.41  June 5  2  34  62.54  3.65  9.74  June 29  4  34  55.48  7.18  16.13  J u l y 19  8  34  61.84  3.43  8.850  Mar. 28  9  53  25.34  0.098  June 5  2  53  70.22  3.90  13.09  June 29  4  53  59.01  7.50  18.30  0.131  w  CQ  July 19  2  53  66.58  4.93  14.74 *  TABLE V (CONT'D) AREA I CAROTENE IN SAMPLES Species S a l i x sp. [Upland  Palatabili t y Rating IA  willow)  Populus tremuloides (Aspen]  Populus t r i e h o o a r p a (Black poplar)  3  3  Date  Weeks i n Sample P e r c e n t Storage No. M o i s t u r e  Mgms. p e r 100 gms Dry Wet  Mar. E8  9  56  41.92  0.71.  1.22  June 5  2  56  72.84  2.70  9.94  June 29  4  56  61.52  3.90  10.14  J u l y 19  1  56  61.93  4.00  10.51  Mar. 28  9  31  43.19  0.42  ' 0.74  June 5  3  31  75.20  3.63  14.64  June 29  4  31  55.95  8.54  19,39  J u l y 19  2  31  59.09  4.69  11.46  Mar. 28  9  33  36.00  0.38  0.59  June 5  3  33  75.07  2.95  11.83  June 29  4  33  68.71  7.10  22.69  J u l y 19  2  33  76.40  3.85  16.31  TABLE V (CONT'D) AREA I CAROTENE IN SAMPLES Species C o>ry, rylus Hazel)  Palatabili t y Rating 4  Date  Weeks i n Sample P e r c e n t Storage No. M o i s t u r e  Mgms. p e r 100 gms Dry Wet  Mar. 28  9  36  30.52  0.30  0.43  June 5  3  38  71.77  2.53  ' 8.96  June 29  4  38  53.32  7.10  15.21  J u l y 19  1  38  61.98  3.68  9.68  6  TABLE VI AREA I I CAROTENE IN SAMPLES Species Betula papyrifera (Paper birch)  Amelanchier f l o r i d a (Service berry)  Cornus s t o l a n i f e r a (Red Osier dogwood)  Falatablli t y Rating 1  1  1  Date  Weeks i n Sample Percent No. Moisture Storage  Mgms. per 100 gms. Dry Wet  Mar. 28  8  40  32.28  0.49  0.72  June 5  4 .  40  78.63  5.90  27.61  June 29  4  40  71.64  9.45  33.32  July 19  1  40  76.34  6.56  27.74  Mar. 28  8  41  27.61  2.65  3.66  June 5  4  41  77.56  1.95  8.68  June 29  4  41  60.92  7.99  20.44  J u l y 19  1  41  63.29  7.05  19.20  Mar. 28  8  54  32.26  2.31  3.41  June 5  4  54  76.39  4.29  18.17  June 29  4  54  63.44  10.77  29.46  July 19  3  54  68.42  9.40  29.77  TABLE 71 (CONT'D) AREA I I CAROTENE IN SAMPLES Species S a l i x sp. (Upland  IA willow)  PopuJ.us t r e m u l o i d e s (Aspen)  Corylus (Hazel)  Palatabili t y Rating  Date  Weeks i n Sample P e r c e n t Storage No. M o i s t u r e  Mgms-. p e r 100 gms. Wet Dry  Mar. 28  8  57  37.25  0.70  1.12  June 5  4  57  75-. 56  3.34  13.67  June 29  4  57  63.23  8.33  22.65  J u l y 19  3  57  68i40  6.57  20-.79  Mar; 28  8  39  33.12  1.08  1.61  June 5  3  39  76*49  4.03  17.14  June 29  4  39  61; 06  8.31  21.34  J u l y 19  1  39  63.05  9.30  25.17  Mar. 28  9  • 45  29.63  2.21  3.14  June 5  3  45  76.44  7.30  20.98  June 29  4  45  59.72  7.48  18.. 57  J u l y 19  2  45  65.67  7.48  21.79  TABLE V I I AREA I I I CAROTENE IN SAMPLES  Species Betula papyrifera (Paper birch]  Amelanchier f l o r i d a (Service berryJ  Cornus s t o l a n i f e r a (Red Osier dogwood)  Palatabili t y Rating 1  1  1  Date  Weeks i n Sample Percent Storage No. Moisture  Mgms. per100 gms. Dry Wet  Mar. 28  7  47  33.24  0.24  0.36  June 5  1  47  79.98  5.38  26.87  June 29  5  47  67.99  8.06  25.18  J u l y 19  3  47  80.85  5.21  27.19  Mar. 28  8  48  34.30  0.304  June 5 '  3  48  78.36  4.84  22.37  June 29  5  48  67.27  12.90  39.41  J u l y 19  3  48  68.24  8.50  26.76  Mar. 28  9  55  31.03  0.45  0.65  June 5  2  55  72.42  6.58  23.86  0.463  ft  June 29  5  55  63.79  14.55  40.18  July 19  1  55  72.91  7.23  26.69  TABLE VII (CONT'D) AREA I I I CAROTENE IN SAMPLES Species S a l i x sp. [Upland willow)  Populus tremuloides (Aspen")  Corylus (Hazel)  Palatabili t y Rating  Date  Weeks i n Storage  Sample Percent No* Moisture  Mgms. per 100 gms Wet Dry  IA  Mar* 28  8  58  34*30  0.54  0.82  June 5  2  58  79.94  7.95  29.63  June 29  4  58  66.03  9.69  28.53  July 19  2  58  67.64  5.95  18.39  Mar: 28  8  46  37:37  0:315  0:503  June 5  3  46  80.18  1:78  8.98  June 29  4  46  81.17  3.63  19.28  July 19  3  46  72.21  3.375  12.14  Mar: 28  9  52  35.97  0.385  June 5  2  52  76.73  5.85  25.14  June 29  5  52  58.78  16.69  40.48  July 19  • 3  52  67.59  8.175  0.601  25.22  TABLE V I I I CHECK AREA ASCORBIC ACID IN SAMPLES Palatabili t y Rating  Speoies Betula papyrifera (Paper b i r c h )  1 •  Amelanchier f l o r i d a (Service berry)  Cornus s t o l a n i f e r a (Red O s i e r dogwood)  1  1  Date  Weeks i n Sample P e r c e n t Storage No. M o i s t u r e  Mgms. p e r 100 gms. Dry Bet  Mar. 28  9  9  36*84  35.84  56.74  June 5  4  9  74.72  99.73  394.50  June 29  3  9  73.51  171.75  648.35  J u l y 19  2  9  73.45  119.80  451.22  Mar. 28  8  3  27.85  32.80  45.46  June 5  2  3  74.90  16.33  65.05  June 29  3  3  59.56  89.40  221.06  J u l y 19  2  3  66.54  131.93  394.29  Mar. 28  8  1  26.22  42.44  57.52  2  1  72.87  69.15  254.88  June 29  3  1  70.62  111.01  377*84  J u l y 19  2  1  68.26  70.73  222.84  June 5  TABLE T i l l  (CONT'D)  CHECK AREA ASCORBIC ACID IN SAMPLES Speoies S a l i x sp. Jpland w i l l o w )  Betula glandulosa CBog b i r o n )  S a l i x sp.  Tswamp willow)  Palatabili t y Rating IA  2  2  Date  Weeks i n Sample Peroent Storage No. M o i s t u r e  Mgms. p e r 100 gms Dry Wet  Mar. 28  9  7  42,87  29.68  51.95  June 5  2  7  72.88  88.93  327.91  June 29  5  7  42.77  162.98  284,78  J u l y 19  2  7  67.24  62.67  191.30  Mar. 28  9  16  20.02  17.51  21.89  June 5  3  16  74.14  58.18  224.98  June 29  3  16  66.87  98.67  297.82  J u l y 19  2  16  68.33  143.87  454.27  Mar. 28  9  12  41.82  49.72  85.45  June 5  2  12  78.48  93.72  435.50  June 29  3  12  66.86  104.42  315.08  J u l y 19  2  12  66.07  197.57  582.29  TABLE V I I I (CONT'D) CHECK AREA ASCORBIC ACID BT SAMPLES Species Populus t r e m u l o i d e s (Aspen)  Populus triehooarpa, (Black poplar)  Corylus Hazel)  PalatabilWeeks i n i t y R a t i n g ' Date Storage 3  3  4  Sample P e r c e n t No. Moisture  Mgms. p e r 100 gms Wet Dry  Mar. 28  8  4  41.94  32.50  55.97  June 5  2  4  72.31  106.25  383.74  June 29  3  4  52.09  144.13  248.89  J u l y 19  .2  4  55.64  214.53  483.61  Mar. 28  9  15  40.43  38.90  65.30  June 5  4  15  71.43  95.48  334.19  June 29  3  15  61.60  198.88  517.91  J u l y 19  2  15  72,77  128.80  473.00  Mar. 28  9  5  34.54  20.60  31.46  June 5  2  5  71.19  89.10  309.26  June 29  3  5  64.49  76.15  214.38  J u l y 19  2  5  65.28  150.10  432.31  TABLE T i l l  (CONT'D)  CHECK AREA ASCORBIC ACID IN SAMPLES Palatabili t y Rating  Speoies LUS oontorta TTTackpine)  latifolia  Pseudotsuga t a x i f o l l a (Douglas f i r )  P i c e a engelmanni ("Spruce)  5  5  6  Date  Weeks i n Sample P e r c e n t Storage No, M o i s t u r e  Mgms. p e r 100 gms. Wet Dry  Mar-. 28  9  18  49.32  236.75  467.14  June 5  2  18  59*99  34.88  87.15  June 29  3  18  64.73  43.83  124,26  J u l y 19  2  18  63.29  96.13  321.61  Mar. 28  8  2  48.53  282.20  548.28  June 5  2  2  59.88  93.65  233.42  June 29  3  2  41.90  82.83  142.57  J u l y 19  2  2  59.64  123.67  306.42  Mar, 28  9  17  17.74  271.50  330.05  June 5 ~  1  17  ' 60.81  86.23  220.03  June 29  3  17  64.58  68.03  192.06  J u l y 19  2  17  63 29  79.07  215.39  •  r  00  TABLE IX AREA : ASCORBIC ACID : I T Speeies Betula papyrifera IPaper birchJ  Palatabili t y Rating  Date  1  Mar* 28 June 5 June 29 J u l y 19  Amelanchier f l o r i d a ~ (Service berry)  1  Mar; 28 June 5 June 29 J u l y 19  Cornus s t o l a n i f e r a (Red Osier dogwood)  1  Mar. 28 June 5 June 29 July 19  S A M P L E S  Weeks i n Sample Percent No. Moisture Storage  Mgms. per 100 gms. Dry Wet  9  52  27.28  45.58  62.40  5  32  74.75  77.88  508.19  4  52  59.62  209.65  521.75  2  52  74.64  187.25  758.29  9  54  58.57  45.65  74.28  2  54  62.54  88.92  257.57  4  54  55.48  129.75  291*44  2  54  61.24  196;55  506-. 55  9  55  25.54  38.66  51.78  2  55  70.22  90.25  502.99  4  55  59.01  158.60  268.72  2  53  66.58  178.45  555.90  TABLE IX (CONT'D) AREA I ASCORBIC ACID IN SAMPLES Species S a l i x sp. [Upland willow)  Populus tremuloides ( Aspen]  Populus triehooarpa (Black poplar)  Palatabili t y Rating IA  3  3  Date  Weeks i n Sample Percent Storage No. Moisture  Mgms. per 100 gms. Wet Dry  Mar. 28  9  56  41.92  49.63  85.45  June 5  2  56  72.84  59,02  217,30  June 29  4  56  61.52  158.60  412.16  July 19  1  56  61.93  103.13  270.90  Mar. 28  9  31  43.19  39.48  69.49  June 5  3  31  75.20  130.58  526.53  June 29  4  31  55.95  208,13  472.48  July 19  2.  31  59.09  337.50  824.98  Mar. 28  9  33  36.00  39.25  61.33  June 5  3  33  75.07  102.00  409.15  June 29  4  33  68,71  210.75  673.53  J u l y 19  2  33  76.40  201.87  855.38  o  TABLE I X (CONT'D) AREA I ASCORBIC ACID IN SAMPLES PalatabilSpecies Corylus  Hazel)  r  4  Date  Weeks i n Sample P e r c e n t Storage No. M o i s t u r e  Mgms. p e r 100 gms Wet Dry  Mar. 28  9  38  30.52  20.13  28.97  June 5  3  38  71.77  72.15  255.58  June 29  4  38  53.32  267.50  573.05  J u l y 19  1  38  61.98  217.17  571.20  TABLE X AREA I I ASCORBIC ACID I N SAMPLES Species Betula papyrifera (Paper b i r c h )  Amelanchier f l o r i d a i Service berry)  Cornus s t o l a n i f e r a (Red O s i e r dogwood)  Palatabili t y R a t i n g • Date 1  1  1  Weeks i n Sample P e r o e n t Storage No. Moisture  Mgms. p e r 100 gms Wet Dry  M a r . 28  8  40  32.28  24.17  35.69  June 5  4  40  78.63  104.18  487.50  J u n e 29  4  40  71.64  209.63  739.17  J u l y 19  1  40  76.34  135.50  572.69  M a r . 28  8  41  27.61  42.77  59.08  June 5  4  41  77.56  43.40  193.40  J u n e 29  4  41  60.92  141.58  361.77  J u l y 19  1  41  63.29  113.40  308.82  M a r . 28  8  54  32.26  59.73  88.18  June 5  4  54  76.39  58.67  248.50  June 29  4  54  63.44  171.43  270.22  J u l y 19  3  54  68.42  136.17  431.19  -  TABLE X (CONT'D) AREA I I ASCORBIC ACID IN SAMPLES Species S a l i x sp. (Upland w i l l o w )  Populus tremuloides (Aspen)  Corylus (Hazel)  Palatability Date Rating IA  3  4  Weeks i n Sample P e r c e n t No. M o i s t u r e Storage  Mgms. p e r 100 gms. Wet Dry  Mar. 28  8  57  37.25  41.35  65.90  June 5  4  57  75.56  90.83  371.64  June 29  4  57  63.23  113.92  309.82  J u l y 19  3  57  68.40  88.13  278.89  Mar. 28  8  39  33.12  28.49  42.59  June 5  3  39  76.49  113.23  481.42  June 29  4  39  61.06  295.00  757.58  J u l y 19  1  39  63.05  306.50  829.50  Mar. 28  9  45  29.63  34.17  48.55  June 5  3  45  76.44  73.55  312.18  June 29  4  45  59.72  189.45  470.33  J u l y 19  2  45  65.67  196.33  571.89  TABLE XI AREA I I I ASCORBIC ACID IN SAMPLES Species Betula papyrifera (Paper b i r c h )  ATTIRI a n c h l e r  (Service  florida Derry;  Cornus s t o l a n i f e r a (Red O s i e r dogwood)  Palatabili t y Rating 1  1  1  Weeks i n Sample P e r c e n t Storage No* M o i s t u r e  Date  Mgms. p e r 100 gms Wet Dry  Mar: 88  7  47  33.24  22.38  33i52  June 5  1  47  79.98  90.38  451.44  June 29  5  47  67.99  83.33  260.32  J u l y 19  3  47  80.85  92.80  484*60  Mar:  8  48  34.30  36:14  55.01  June 5  3  48  78.36  24.10  111.37  June 29  5  48  67.27  103.08  314.94  J u l y 19  3  48  68.24  76.83  241.91  Mar. 28  9  55  31.03  32.84  47.61  June 5  2  55  72.42  54.22  196.59  June 29  5  55  63.79  127.55  352.17  J u l y 19  1  55  72.91  107.30  396.09  28  -  •  •  TABLE I T . (CONT'D) AREA I I I ASCORBIC ACID IN SAMPLES Species S a l i x sp.  itabilRating IA  Copland willow)  Populus t r e m u l o i d e s (Aspen)  Corylus (Hazel)  3  4  •  Date  Weeks i n Sample P e r c e n t Storage No. Moisture  Mgms. p e r 100 gms Wet Dry  Mar. £8  8  58  34.30  41.12  62.59  June 5  2  58  79.94  57.25  285.39  June £9  4  58  66.03  99.90  294.08  J u l y 19  2  58  67.64  82.47  254.85  Mar. 28  8  46  37.37  27.03  43.15  June 5  3  46  80.18  91.67  462.51  June 29  4  46  81.17  138.83  734.62  J u l y 19  3  46  72.21  162.43  584.49  Mar. 28  9  52  35.97  25.10  39.20  June 5  2  52  76.73  54.72  235.15  June 29  5  52  58.78  143.83  348.93  J u l y 19  3  52  67.59  83.83  258.58  •  56.  TABLE X I I EFFECT OF FREEZING ON CAROTENE Pinus oontorta yar. l a t i f o l i a  Date  Peroent  (Jackpine)  Moisture  Mgms. p e r 100 gms. Wet Dry  June 3  65.47  3.15  9.12  June 17  65.32  3.00  8.65  July 1  60.59  3.34  8.47  July 8  60.32  3.33  8.40  J u l y 22  62.35  3.14  8.33  Aug. 5  60.08  3.26  8.17  atula p a p y r i f e r a (Paper b i r c h )  Date  Percent Moisture  Mgms. per 100 gms. Wet Dry  June 3  47; 11  18.18  34.37  June 17  51.82  15.73  32.65  July 1  35.57  20.72  32.16  July 8  40.27  18.98  31.78  J u l y 22  69.68  9.82  32.39  Aug. 5  71.96  8.65  30.85  TABLE X I I I EFFECT OF FREEZING ON ASCORBIC ACID  Pinus oontorta var. l a t i f o l i a Date  (Jaokpine)  Peroent M o i s t u r e  Mgms. p e r 100 gms Dry Wet  June 3  65.47  47.07  165.28  June 17  65.32  53.63  154,64  July 1  60.59  63.87  162.08  July 8  60.32  63.12  159.07  J u l y 22  62.35 .  56.87  151.04  Aug. 5  60.08  63.60  151.32  J e t u l a p a p y r i f e r a (Paper b i r c h ) Date  Peroent Moisture  Mgms. p e r 100 gms. Wet Dry  June 5  47.11  121.72  230.13  June 17  51.82  103.57  214.96  July 1  35.57  139.33  216.25  July 8  40.27  130.02  217.68  J u l y 22  69.68  65.65  216.52  August 5  71.96  64.30  219.31  TABLE X I V PALATABILITY RATING AND TOTAL VITAMIN CONTENT Palatabili t y Rating  Species  Carotene Check All Area Areas  B e t u l a p a p y r i f e r a (Paper birch.)  1  65.86 286.93  Amelanchier  1  Ascorbic Aoid Check All Area Areas 1550.81  6246,35  75.63 251.743  725.86  3481.78  1  79.96 298.411  913.08  4101.02  S a l i x sp. (Upland w i l l o w )  IA  52.94 220.35  855.94  3764.91  B e t u l a g l a n d u l o s a (Bog b i r c h )  2  32.71  998.96  S a l i x sp. (Swamp w i l l o w )  Z  49.29  1418.32  Populus  3  62.57 214.963 1172.21  Populus triehooarpa (Black poplar)  3  59.28  Corylus  4  63.50 256.67  f l o r i d a (Service berry)  Cornus s t o l a n i f e r a  (Red O s i e r dogwood)  t r e m u l o i d e s (Aspen)  (Hazel)  7001.55  1390.40 987.41  4701.02  59.  FIGfD'HE 4  AREA  I  Area I I I (Area I In background)  63.  FIGURE 8 EFFECT OF STORAGE,  Carotene  Birch  Pine  3  June  17 June  1 July  8 July  22 July  5  Aug,  FIGURE 9 EFFECT OF STORAGE,  3 June  17 June  1 July  Ascorbic Acid  8 July  22 July  5 Aug.  FIGURE 10 CAROTENE CALIBRATION CURVE  66 FIGURE  11  ASCORBIC ACID CALIBRATION 'S'  -  AND  "SFECTflOPKOTELPMETERS"  Transmittaiicy  CURVE  67.  FIGURE. 12 COMPARISON OF AREAS - CAROTENE - JUNE 5  Betula papyrifera  ii in  Amelanchier florida  il !tl CA  Cornus Stolanifera  II III C  S a l i x sp. [Upland w i l l o w )  A  li  in CA  Populus remuloides Populus riohooarpa  Corylus  II  in  ca  [IE3 * a  II  \ IL 1 AvcoU  CP)  II  | J X | Areom  i  la 10  20  30  mgms./ 100 gms.  68.  FIGURE 13 COMPARISON OF AREAS - CAROTENE - JUNE 29  Betula  C  A.  papyrifera Ill  Amelanchier florida  CA  Cornus stolanifera  S a l i x sp. (Upland willow CA  Populus remuloides  Populus trichooarpa  CA  CA  Corylus III  10  20  30  mgms./ 100 gms.  69 FIGURE 14 COMPARISON OF AREAS - CAROTENE - JULY 19  CA  Betula papyrifera  Amelanchier florida  Cornus stolanifera  S a l i x sp. [Upland willow)  Populus tremuloides Populus trichooarpa  CA  C-A  Corylus  i l  1 lI  0  10  20  30  mgms*/ 100 gms.  FIGURE 15 COMPARISON OF AREAS - ASCORBIC ACID - JUNE 5  C  A  Betula papyrifera III  Amelanchier florida  CA  Cornus stolanifera  M in  i  1  M S a l i x sp. (Upland willow)  i  I  3  III  Populus remuloides  -Populus trichooarpa  CA  CA  Corylus 15  0  200  40-0  mgms./ 100 gms.  71. FIGURE 16 COMPARISON OF AREAS - ASCORBIC ACID - June 29  Betula papyrifera  Amelanchier florida  Cornus stolanifera  S a l i x sp. (Upland willow)  Populus 3 remuloides  Populus trichooarpa  Corylus  0  200  400  600  mgms./ 100 gms.  FIGURE 17 COMPARISON OF AREAS - ASCORBIC ACID - JULY 19  200  400  600  800  mgms./ 100 gms.  73. LEGEND - FIGURES 18-25 1  .-  Betula papyrifera  2 --  Amelanchier  3 --  Cornus s t o l a n i f e r a  4 •-  S a l i x sp. (Upland willow)  florida  5 -- Populus tremuloides 6 --  Corylus  FIGURE 18 SEASONAL VARIATION - DECIDUOUS SPECIES CAROTENE - CHECK AREA  28 Mar.  5 June  29" 19 Jump . July  FIGURE S E A S O N A L  19  V A R I A T I O N  -  D E C I D U O U S  CAROTENE  -  A R E A  I  S P E C I E S  75.  FIGURE EO SEASONAL VARIATION / DECIDUOUS SPECIES CAROTENE - AREA  E8 Mar.  U  5  June  S9 June  19 J uly  76, FIGURE S I SEASONAL  Mar.  VARIATION  -  DECIDUOUS  CAROTENE  -  AREA  SPECI3S  III  June  June  July  FIGURE EE SEASONAL VARIATION - DECIDUOUS SPECIES ASCORBIC ACID - CHECK AREA  28 Mar  5 June  £9 J une  19 July  FIGURE 23 SEASONAL VARIATION - DECIDUOUS SPECIES ASCORBIC ACID - AREA I  FIGURE 24 SEASONAL VARIATION - DECIDUOUS SPECIES ASCORBIC ACID - ABBA I I  FIGURE 25 SEASONAL VARIATION - DECIDUOUS SPECIES ASCORBIC ACID - AREA I I I  28  Mar.  • 5 June  2,9 J une  19 July  81. FIGURE 86 SEASONAL VARIATION - CONIFERS CAROTENE - CHECK AREA  o A •  88 Mar.  5 June  o  Pinus oontorta  A  Pseudotsuga  a  P i o e a engelmanni  taxifoila  89 June  19 July  FIGURE 27 SEASONAL VARIATION - CONIFERS ASCORBIC ACID - CHECK AREA  1  :  28 Mar,  '  5 June  o  Plnus oontorta  A  Pseudotsuga t a x i f o l i a  D Picea engelmanTil  i  29 June  i  _  19 July  7  

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