COBALT POLYCYTHEMIA IN CATTLE By George M e r r i l l Ward A THESIS Subm itted t o t h e School o f G raduate S tu d ie s o f Michigan S ta t e C o lleg e o f A g r i c u ltu r e and Applied Science in p a r t i a l f u l f i l l m e n t o f t h e r e q u ire m e n ts f o r th e d eg ree o f DOCTOR OP PHILOSOPHY Department o f D airy ACKNCWLPDGMSHTS The w r i t e r w i^ i e s to e x p r e s s h i s s in c e r e a p p r e c i a t i o n to Doctor C. P . Huffman, R esearch P r o f e s s o r o f D airying, f o r h i e co u n sel m d t i n e l y s u g g e s tio n s th ro u g h o u t t h i s s tu d y and f o r th e c r i t i c a l r e r d i n g o f th e m a n u s c rip t; t o Doctor S a r i Weaver, P r o f e s s o r o f D airy in g , f o r t h e av;erd of th e G raduate A s s i s t n n t ship and f o r th e n r o v i s i o n o f th e f a c i l i t i e s r e q u i r e d f o r th e consummation o f t h i s s t u d y . The w r i t e r i s in d e b te d t o Mr. C. i.'. Duncan, R esearch A s s o c ia te in A-gri c u l t u r a l Chem istry, and h i s a s s o c i a t e s f o r th e chem ical a n a l y s e s u s e d h e r e i n ; and to unnamed members o f th e D airy Department f o r t h e i r many s u g g e s tio n s r e g a r d i n g the conduct o f the study an a th e p r e p a r a t i o n o f th e m a n u s c rip t. The w r i t e r w ish es to e x ten d g r a t i t u d e to h i s w if e whose n e v e r -e n d in g encouragement h a s been i n v a l u a b l e . TABLE OF CONTENTS INTRODUCTION..................................................................................................... REVIEW OF LITERATURE................................................................................... I n t r o d u c t i o n .......................................................................................... The E s s e n t i a l i t y o f C o b a l t ........................................................... Mode o f A ctio n o f Cobalt i n R u m in an ts.................................. C o b a lt-In d u c e d P o ly cy th em ia— I n c i d e n c e ................................ F a c t o r s A f f e c t i n g CobPlt P o ly c y th e m ia .................................. Mechanism o f Cobalt P o ly c y th e m ia ............................................. Cobalt T o le ra n c e i n R u m in an ts.................................................... S u lf a g u p n id in e ...................................................................................... EXPERIMENTAL PROCEDURE............................................................................... RESULTS................................................................................................................. Blood P i c t u r e ........................................................................................ Growth........................................................................................................ DISCUSSION.......................................................................................................... SUMMARY AND CONCLUSIONS............................................................................. LITERATURE CITED............................................................................................ INDEX OP FIGURES P ig u re 1 . Blood p i c t u r e and growth curve o f anim al number C680 F ig u re c • Blood p i c t u r e and growth curve o f animal numbe r 0681 F ig u re 3- Blood p i c t u r e ana growtv curve o f anim al number 068*4 F ig u re *4. Blood p i c t u r e rnd growth cu rv e o f a ni mal number 0718 P ig u re 5- Blood p i c t u r e and g rrw th curve of anim al number C719 F ig u re 6 . Blood p i c t u r e ana growth curve o f an? mal number 072*4 F ig u re 7- Blood ■oicture and grow ti t c u rv e o f animal number 07*4-2 F ig u re 3 . Blood p i c t u r e and growth curve o f animal number 07*49 F ig u re 9- Blood p i c t u r e and growt'-' curve o f animal number 0715 P ig u re 1 0 . Blood p i c t u r e and growth curve o f a n i ’-al number C739 P ig u re 1 1 . Blood p i c t u r e and grov/th c u r v e o f animal. numbor 07*41 F ig u re 1 2 . Blood p i c t u r e end growth curve of animal number C7*r*4 F ig u re 13. Blood p i c t u r e ana growth c u rv e o f anir.nl number 0711 P ig u re 1*4. Blood p i c t u r e and growth curve o f anim al number 0725 P ig u re 15- Blood p i c t u r e end growth curve of animal number 0730 P ig u re 1 6 . Blood p i c t u r e and growth curve o f animal number 071*4 Pi gure 17- Blood p i c t u r e end growth curve o f anim al number 0726 P ig u re 1 8 . 31ood p i c t u r e o f anim al number A78 P ig u re 19- Blood p i c t u r e o f a n irip l number A77 Pi gure 2 0 . Comparison o f growth o f an im al'? in: Groups 1, 2, 3, and 4 w ith t h e R ag sdale s ta n d a r d F ig u re 2 1 . Comparison o f growth o f an im als in Groups 5 w ith th e R agsdale s ta n d a rd £ I1ID3X OF TABLES Table 1 . Composition o f E x p erim en tal Groups Table 2. Key t o A b b re v ia tio n s Used on F i b r e s Appendix Table 1 . Blood n i c t u r e o f anim al number 0680 Appendix T able 2. Blood p i c t u r e o f anim al number 0681 Apoendix Table 3* Blood p i c t u r e o f a n i r n l number C<'34 Appendix Table 4 . 31ood p i c t u r e o f anim al number C718 Appendix Table $. Blood n i c t u r e o f a n 'n n l number 0719 Appendix Tnble 6 . Blood p i c t u r e o f a n ir.n l number 0724 Appendix T ab le 7* Blood p i c t u r e o f anim al number 0742 Appendix Table 8 . Blood -n ic tu re o f anim al number C7*J-9 Apnendix Table 9» Blood p i c t u r e o f anim al number C715 im pendix Table 1 0 * Blood n i c t u r e o f anim al number C739 Appendix T able 1 1 . Blood p i c t u r e o f anim al number C7*4l Appendix Table 1 2. Blood -n ic tu re o f animal number 0744 Appendix Table 13* Blood p i c t u r e o f anim al number 0711 Appendix Table 14. Blood p i c t u r e o f anim al number C725 Appendix T able 15* Blood p i c t u r e o f anim al number C730 Appendix T able 1 6 . Blood p i c t u r e o f anim al number 0714 Appendix Table 17* Blood p i c t u r e o f anim al number 0726 Appendix Table 1 8 . 31ood p i c t u r e o f anim al number A73 Appendix T able 19* Blood p i c t u r e o f anim al number A75 Appendix T able 20. 3 lo od n i c t u r e o f anim al number A?8 Appendix Table 21. Blood p i c t u r e o f anim al number ASO iiidsx op tables - co^ t i i u s d Page Appendix Tnble 2 ?. Blood n i c t u r e o f anim al number A72 96 Appendix Table 23. Blood p i c t u r e o f animal number A76 97 Appendix Table 2b. Blood p i c t u r e o f on:'mal number A77 98 Apnendix Table 2 5 . Blood p i c t u r e o f an-'mal number A79 99 Apnendix Table 2 6 . Growth r e c o r d Apnendix Table 27- Comparison o f body weight w ith the Ragsdale S tan dard— Grouns 1, 2, 3. Appendix Table 28. 100 ** 106 Comparison o f body w eight w ith th e R ag sd ale S tand ard — Grouns 5 and 6 107 JK 1 INTRODUCTION P r e s e n t knowledge i n d i c a t e s t h a t c o b a l t in t r a c e q u a n t i t i e s i s e s s e n t i a l f o r th e w e ll b e in g o f r u m in a n ts . V ariou s "w asting d i s e a s e s " o c c u r r i n g i n l o c a l i z e d a r e a s o f th e w orld have been cured and subse­ q u e n tly p r e v e n t e d by p r o v i d i n g ru m in an ts w ith a c c e s s to t h i s elem ent by any of s e v e r a l means, i n c l u d i n g p r o v i s i o n o f c o b a l t l i c k s , f e e d in g c o b a l t i z e d s a l t , and i n c r e a s i n g the c o b a l t c o n te n t o f fo ra g e by to p d re B s in g w i th c o b a lt s a l t s . The manner i n which c o b a l t e x e r t s i t s e f f e c t on ru m in an ts h a s n o t b -e n e l a b o r a t e d . The e s s e n t i a l i t y o f c o b a l t , i n a d d i t i o n to t h a t c o n ta in e d in Vitam in B ^ , h a s n o t been d e m o n stra te d f o r any group o f a n im a ls o t h e r th an r u m in a n ts . C onversely, up to t h i s tim e, i t h a s not been ^iown t h a t c o b a l t in q u a n t i t i e s g r e a t e r th an r e q u i r e d f o r t h e i r w e ll b e in g i s a b le t o su pp ort p o ly c y th e m ia in ru m in a n ts as i t i s in the c a s e s o f o t h e r a n im a ls . These a p u a re n t d i f f e r e n c e s i n the a c t i o n o f c o b a l t on th e v a r i o u s an im als r a i s e th e q u e s ti o n s o f how th e y o ccu r and what p h y s io lo g ic a l v a r ia tio n i s re sp o n sib le fo r th ese d if f e r e n c e s . T h is i n v e s t i g a t i o n was i n i t i a t e d in an a tte m p t to a s c e r t a i n w h ether or not i t is p o s s ib le to produce a c o b a l t - in d u c e d p o ly c y th e m ia i n c a t t l e and i n c o n ju n c tio n w i t h t h a t to a tte m p t to determ in e the fun c­ t i o n o f th e rumen m icroorganism s i n the r u m i n a n t's u s e o f c o b a l t by u t i l i z i n g s u lf a g u a n i d i n e a s a b a c t e r i o s t a t i c a g e n t. - 2 - REVIEW OF LITERATURE I n tr o d u c t i o n The l i t e r a t u r e r e g a r d i n g th e e s s e n t i a l i t y o f coha.lt end t h e e f f e c t t h a t t h i s elem ent may e x e r t on th e b lo o d o f a n im a ls , w i t h few e x c e p t i o n s , h a s been p r i n t e d w i t h i n th e l a s t tw enty-one y e a rs* In 1935 t h e e s s e n t i ­ a l i t y o f c o b a lt f o r rum inant n u t r i t i o n was d is c o v e r e d in c o a s t a l r e g i o n s of A u s tra lia .. In 1929 i t was d is c o v e r e d t h a t c o b a l t s a l t s , when ad m in is­ t e r e d in q u a n t i t i e s g r e a t e r th a n n o rm a lly found in r a t i o n s , cau ses a p o ly cy th em ia in r a t s . Since th e s e d i s c o v e r i e s were made many c o n t r i b u ­ t i o n s hove been made to th e l i t e r a t u r e co n c e rn in g th e e f f e c t s o f v a r i o u s l e v e l s o f c o b a l t a d m i n i s t r a t i o n to d i f f e r e n t s p e c ie s o f a n im a ls. The E s s e n t i a l i t y o f C o b alt ’ Anemia and e m a c ia tio n have c h a r a c t e r i z e d c e r t a i n "w asting d i s e a s e s " which have a f f e c t e d c a t t l e and sheep in many p a r t s o f th e world f o r many years. Cobalt su p p le m e n ta tio n has been shown to cure and p r e v e n t "Bush- Siclcness" and "M ortain Mains D is e a se " i n New Zealand ( b ) , in New Zealand (3 8 ), "Mairoa D opiness" "E nzootic Marasmus" i n A u s t r a l i a (1 3 0 ), " N u t r i t i o n a l Anemia" in New South Wales ( 5 9 ), " N a k u r u i tis " i n Kenya ( 2 ) , G reat B r i t a i n (3 l)» "P in in g 11 in "Grand T ra v e rse D is e a s e " in M ichigan ( 6 ) , " s a l t s ic k " in F l o r i d a (92, 9 3 ), c o b a l t d e f i c i e n c y i n W isconsin (52) and New York (1 17 ), " B u r to n - a il" in New Hampshire (6 9 ), and d e f i c i e n c i e s i n th e H e b rid e s ( l 6) and Norway (b?)* Loss o f w eight and a p p e t i t e , g e n e r a l u n t h r i f t i n e s s , - 3 weakness, anemia, end f i n a l l y d e a th c h a r a c t e r i z e a l l th e s e d i s e a s e s which have been r e p o r t e d . The l i t e r a t u r e co n c e rn in g c o b a l t d e f i c i e n c i e s h a s been amply review ed in r e c e n t y e a r s (11, 62, 10B, 115, 1 2 8 ). C obalt s u p p le m e n ta tio n h a s been shown t o be ad v an tag eo u s u n d e r co n d i­ t i o n s o f no o bv iou s d e f i c i e n c y a i l m e n t . K losterm an et, a l . (75) have demon­ s t r a t e d t h a t f a t t e n i n g swine m a in ta in e d in c o n c r e te p en s which were washed d a i l y made s i g n i f i c a n t l y g r e a t e r d a i l y g a i n s v;hen c o b a lt c h l o r i d e was added a t the r a t e o f 0 .0 2 6 u e r cen t to th e r a t i o n . Noland e_t a l . (9*0 ob­ t a in e d f a v o r a b le growth r e s u l t s w ith a supplement c o n s i s t i n g o f c o b a l t and copper s a l t s when added to an a l l - p l a n t r a t i o n f o r growing p i g s . Supple­ m en tatio n o f th e a l l - p l a n t r a t i o n w i t h i r o n , manganese, c o b a l t , and copper su p p o rted a much g r e a t e r c o n c e n tr a t i o n o f v ita m in B^r, i n the f e c e s than d id su p p lem en tatio n w ith APF supplem ents o r c r y s t a l l i n e v ita m in B ^ . Kato (6 ?) h a s d em o n strated t h a t c o b a l t i s o f v a lu e when u sed in c o n ju n c tio n w ith i r o n th e r a p y in the t r e a tm e n t o f i n f a n t anem ia. Since re q u ire m e n t f o r co­ b a l t by human b e in g s h a s n o t been d e m o n stra te d , t h e a c t i o n e x e r t e d by c o b a l t in t h i s work may w e ll have been t h a t o f p oly cy th em ia p r o d u c t io n . No p o s i t i v e c o b a lt req u irem en t in a d d i t i o n to v ita m in B ^ c o b a lt (107) h a s been d em o n strated f o r a n im a ls o t h e r th an sheep and c a t t l e w i t h the p o s s i b l e e x c e p tio n s n o te d above (75, 9*0. The s p e c i e s d i f f e r e n c e i n c o b a lt req u ire m e n t 1 e i n d i c a t e d by th e f a c t t h a t h o r s e s a r e n o t a f f e c t e d on c o b e l t d e f i c i e n t p a s t u r e s which do n o t f u l f i l l th e n u t r i t i v e r e q u ire m e n ts o f sheep and c a t t l e . Underwood and Elvehjem (l2 9 ) were a b le to m a in ta in growth i n r a t s on a d i e t c o n s i s t i n g o f m ilk f o r t i f i e d w ith i r o n and copper which s u p p lie d only 0 .6 microgrem o f c o b a lt d a i l y . Thompson and E H I r (126) found th e req u irem en t f o r c o b a l t in r a b b i t s t o be l e s s th e n 0 .1 microgram p e r day. Mode o f A ctio n o f C obalt In Ruminants The mode o f a c t i o n o f c o h e lt in ru m in o n ts i s some i n v e s t i g a t o r s have p r e s e n t e d s u g g e s t iv e d a t a . n o t known a lth o u g h The f a c t t h a t non- rum inant anim als have n o t been shown t o r e q u i r e c o b a lt te n d s to n o i n t to th e rumen a s th e organ r e q u i r i n g c o b a l t f o r i t s a c t i o n . Marnton and Lee ( 83) compared th e e f f e c t s o f sub cu taneou s i n ,l e c t i o n and o r a l c o b a l t ad­ m i n i s t r a t i o n on sh eep. Since only o r a l l y a d m in is te r e d c o b a l t s u p o o rte d grow th, th e se w o rkers concluded t h a t c o b a l t " e x e r t s i t s i n f l u e n c e on th e rum inant p r i m a r i l y e i t h e r i n th e lumen o f th e a lim e n ta r y c a n a l o r when p a s s i n g through i t s w a l l . " t h e y reaso n ed t h a t , F u r th e r l i m i t i n g th e s i t e o f c o b e l t a c t i o n , s in c e a p o r t i o n o f th e i n j e c t e d c o b a l t 1b e li m i n a t e d in the f e c e s and on ly a p p e a rs a t th e i n t e s t i n a l l e v e l ( 27) , th e a c t i v i t y a p p a r e n tly t a k e s p l a c e above the l e v e l o f th e duodenum. These i n v e s t i g a ­ t o r s p ro po se t h a t c o b a l t f u n c tio n i n ru m in a n ts may concern t h e a c t i v i t y o f the sy m b io tic rumen f l o r a . V e r i f i c a t i o n o f t h e i r d e d u c tio n s i s p a r ­ t i a l l y e s t a b l i s h e d in t h a t nonrum inants have n o t y e t been shown t o r e q u i r e c o b a lt o th e r th a n t h a t c o n ta in e d i n v ita m in B^g ( 1 0 7 ). Keener a t a l . (68) , s t a r t i n g w ith c o b a l t d e f i c i e n t lambs, were a b le to g e t slow b u t p o s i t i v e re c o v e ry u s in g in tr a v e n o u s c o b a l t i n j e c t i o n s a s compered t o ra n id reco v ery a f t e r o r a l c o b a lt a d m i n i s t r a t i o n . T h is stu d y c o r ro b o r a te d t h e r e s u l t s o f on e a r l i e r study by Roy et, a l . ( 106) . T h is slow improvement moy be accounted f o r b y the c o b a l t c o n te n t o f s a l i v a ( 2 9 ) . C obalt i n j e c t i o n s in l e s s e r q u a n t i t i e s were n o t e f f e c t i v e in a l l e v i a t i n g c o b a l t d e f i c i e n c y in lambs (117)In an a tte m p t t o l i n k volume o f v ita m in Bjp p r o d u c tio n i n th e rumen w ith c o b a l t d e f i c i e n c y in sheep H ale e t a l . (58) d e m o n stra te d t h a t c o b a lt supplem ented sheep su p p o rted much g r e a t e r p r o d u c tio n o f th e v ita m in i n th e rumen then d id c o b a l t - d e f i c i e n t sh eep . The work o f Becker e t a l . (10) e s s e n t i a l l y n u l l i f i e d th e v a lu e o f th e above m entioned work a s i t r e g a r d s th e a c t i o n o f c o b a l t on rum inant a n im a ls . These i n v e s t i g a t o r s were u n a b le to g e t any resp o n se when v ita m in B^j W0B i n j e c t e d i n t r a m u s c u l a r l y tw ice weekly f o r Bix weeks in c o b a l t - d e f i c i e n t lam bs. S im ila rly o ra l a d m in istra­ t i o n o f a v ita m in B ^ c o n c e n tr a t e y i e l d e d no r e s p o n s e . These r e o o r t s seem to e l i m i n a t e v ita m in B^^ p r o d u c t io n a s b e in g th e im p o rta n t u se o f t h e c o b a lt r e q u i r e d by rum inant a n im a ls . T o s ic and M itc h e ll (127) d ete rm in e d th e c o b a l t co n te n t o f rumen- m ic r o b ia l m a t e r i a l from sheep on n a t u r a l hoy r a t i o n s 8nd found ap p ro x im ate­ l y 80 p e r c e n t o f t h e t o t a l rumen c o b a l t c o n ta in e d in th e m ic r o b ia l f r a c ­ tio n . They were a b le to i n c r e a s e th e m ic ro b ia l c o b a l t c o n te n t m arkedly by su pp lem enting th e hay r a t i o n w ith one m illig ra m c o b a l t d o i l y . I t is of i n t e r e s t in t h i s c o n n e c tio n t h a t Pounden e t a l . ( 1 0 3 ), in a etu dy o f th e f a t e o f fo u r ty p e s o f m icroorgen5sm s w hich a r e c h a r a c t e r i s t i c a l l y p r e s e n t in rumen samples ta k e n from c a t t l e , found t h a t t h e s e s e v e r a l ty p e s o f m icroorganism s v ary in t h e i r r e s i s t a n c e to d i g e s t i o n . Some pasB c o u p l e t e ly i n t a c t th ro ug h th e a l i m e n t a r y c e n a l w h ile o t h e r s d is a p p e a r a s e a r l y as th e abomasum. Even though t h e r e i s much y e t to be i n v e s t i g a t e d c o n cern in g th e f e t e o f c o b a l t i n th e rumen and in th e m icroorganism s which p a s s from th e rumen th e l a s t two q uo ted s t u d i e s a r e s u g g e s tiv e o f th e f a t e o f c o b a l t i n rum inan t a n im a ls . ^ - 6 - C o b n lt-In d u ce d P o lycyth em ia— In c id e n c e The a b i l i t y o f c o b a l t to pro du ce p o ly c y th e m ia when A d m in istered i n l a r g e d o ses has r e c e i v e d c o n s id e r a b le A t t e n t i o n . O b se rv a tio n o f t h i s p r o p e r t y was f i r s t mrde by W altner and Y frltner ( 135) w^o f e d r a t i o n s con­ t a i n i n g o n e - h a l f and two p e r c e n t c o b a l t , r e s p e c t i v e l y , to r a t s * The l a r g e r c o b a l t gupplem entnt'.on was r e l a t i v e l y t o x i c w h ile t h e sm a lle r allow ed normal grow th b u t caused i n f e r t i l i t y . Since t h a t time c o b a l t - indnced n o ly c y th e m ia h a s been r e p o r t e d in r a t s ( l , 9* 17, 21, 43, 55. 6 5 , 78, 82, 91, 99, 1 1 9 ,1 2 1 ), r a b b e t s (8, 54, 74. 78, 89, 111, 113, 1 3 4 ), c h ic k e n s ( l l 4 ) , ducks ( 4 2 ), salam an ders (2 5 ), mice ond f r o g s (122) g u in e a p i g s (89, 122, 1 3 l ) , p i g s ( l 2 l ) , dogs ( l 8 , 36, 49, 8 6 ) , and human b e in g s (1 3 ? ), and p o s s i b l y in sh°ep ( 6 6 ). F a c t o r s A f f e c t i n g C o b alt P o ly cy th em ia S te n le y et. a l. (120) have shown t h a t i t tn k e s s i x t e e n tim es th e dose o f c o b a lt a d m in is te r e d o r a l l y to produce th e seme p o ly c y th e m ic e f f e c t in r a t s a s when i t i s a d m in is te r e d s u b c u ta n e o u s ly . depends on i t s r o u te o f a d m i n i s t r a t i o n . The e x c r e t i o n o f c o b a lt Greenberg et, a l . (30, 56) found t h a t o r a l l y a d m in is te r e d c o b a lt i s o n ly p a r t i a l l y abso rb ed, th e rem aind er u l u s p a r t o f t h a t c o n ta in e d in th e b i l e b e in g q u ic k ly e l im in a t e d in the feces. A p o r t i o n o f th e abso rb ed c o b a lt i s accum ulated in th e l i v e r and some i s e li m i n a t e d i n the u r i n e . The r a p i d e l i m i n a t i o n (5, 27, 28) and l i v e r s to r a g e o f c o b a l t (27, 28) have been co n firm ed . Kent and McCance (71) showed t h a t a man e x c r e te d 20 p e r c e n t o f h i s food c o b a l t in th e u r i n e w h ile i n j e c t e d c o b a l t was slow ly e x c r e te d i n the u r i n e . Conor and L e v is (27) d em o n strated t h a t c o b a lt i n j e c t e d i n t o th e j u g u l a r v e in wbb e x c r e te d in th e u r i n e and f e c e s in the ap pro xim ate r a t i o o f two to one. Human b e in g s e x c r e t e 60 to 80 p e r c e n t o f i n j e c t e d c o b a l t in th e u r in e (1 3 7 ). Houk et. al.. (60) showed t h a t r e t e n t i o n o f c o b a l t i n r a t s v a r i e d i n v e r s e l y w ith the dosageGordon et. a l . (55) d e m o n stra te d t h e v a lu e o f c o b a l t a s a supplement f o r th e s t i m u l a t i o n o f r e d b lo o d c e l l and hem oglobin p r o d u c t io n i n b le d th y ro id e c to m iz e d r e t s * even b e t t e r r e s u l t s * The a d d i t i o n o f th y r o x i n t o the supplem ent gave T e s to s te r o n e h as been shown t o be n e c e s s a r y f o r hemoglobin fo rm a tio n (^ 8 ) b u t an e x c e s s o f t h i s hormone i s d e l e t e r i o u s . These f r e t s te n d t o show th e r e q u ire m e n t f o r b a la n c e d hormone p r o d u c tio n f o r e r y t h r o p o i e B is . L iv e r e x t r a c t s have been shown t o a c c e l e r a t e t h e r e t u r n to norm alcy o f c o b a lt- p o ly c y th e m ic r e t s (R2 , 111) and guinea, p i g s ( i l l ) b u t t h e r e was no i n t e r f e r e n c e w ith th e p o ly cy th em ia when b o th were a d m in is te r e d t o g e t h e r . Anderson et. a l . ( l ) r e p o r t e d t h a t e i t h e r whole l i v e r powder o r W ils o n 's l i v e r e x t r a c t a id e d c a b a l t - p o ly c y t h e m ir p r o d u c tio n i n r a t s . Davis ( 37*38) showed t h a t l i v e r re d u c e s p o ly cy th em ia in d o g r. However, i t r e t u r n e d fo u r days a f t e r l i v e r f e e d in g was sto p p ed . Cobalt h a s b -en shown t o i n h i b i t normal h e m a to p o ie tic resu o n se to ir o n and cop per f e e d in g in dogs made anemic by hemorrhpgevitoreas whole d ry l i v e r or l i v e r e x t r a c t a d m i n i s t r a t i o n overcame th e i n h i b i t o r y e f f e c t (50)» B arron ( 7) found only a d e c re a s e d r a t e o f po ly cy th em ia p r o d u c t io n when c o b a lt and sm ell d a i l y i n j e c t i o n s o f L i l l y ' s l i v e r e x t r a c t were a d m in is te r e d t o g e t h e r . C o balt p o ly cy th em ia in dogs h a s been shown t o be d e p r e s s e d by d o i l y a d m i n i s t r a t i o n o f a s c o r b i c a c i d (39) a lth o u g h i t had no e f f e c t on low- p r e s s u r e p o ly c y th e m ia . C o balt was found to reduce the b lo o d a s c o r b i c a c i d l e v e l s o f c o b a lt- p o ly c y th e m ic dogs. A sco rb ic a c id h as been tised to a l l e ­ v i a t e the e f f e c t s o f c o b a l t in r a b b i t s (7. 8 ) and human b e in g s (137)* Levy ( 78) v/as not able to s u b s t a n t i a t e th e s e r e s u l t s w ith r a t s and r a b b i t s b u t th e i n tra v e n o u s a d m i n i s t r a t i o n o f th e sodium s a l t o f a s c o r b i c a c id d e c re a s e d th e hemoglobin c o n c e n t r a t i o n i n th e b lo o d o f p o ly cy th em ic r a b b i t s . The sodium s a l t o f i s o n s c o r b ic a c id had a s i m i l a r b u t l e s s e r e f f e c t and th e sodium s a l t o f g l u c o a s c o r b i c a c i d hod no e f f e c t on the o o ly e y th e m ia . Choline h as been shown to e x e r t no e f f e c t on c o b a l t n o ly c y th e m ia in r a t s ( 2 1 ,2 2 ) . R e tu r n to normal r e d b lo o d c e l l coant was h a s te n e d by c h o lin e a d m i n i s t r a t i o n in r a b b i t s made p o ly cy th em ic w ith c o b a l t ( l i o ) and c h o lin e d e p re ss e d h e ra n to p o ie s is i n c o b a lt- p o ly c y th e m ic dogs (3 8 ). Burke e t a l . (?M) found t h a t mice could t o l e r a t e much more c o b a l t when i t was accompanied by c o n c u rre n t i n j e c t i o n s o f h i s t i d i n e in more th a n equim olar q u a n t i t i e s . G r i f f i t h e t h i . (57) were able e s s e n t i a l l y to p r e v e n t th e i n h i b i t i o n of grow th o f young r a t s due t o c o b a l t a d m in is tr a .tio n by supplem enting t h e r a t i o n w ith m e th io n in e , c y s ti n e o r c y s t e i n e , e s p e c i a l l y th e l a t t e r . These w o rk ers e s t a b l i s h e d th e r e l a t i v e n o n t o x i c i t y o f th e c o b n l t - c y s t e i n e complex formed in v i t r o . Orten and B u cc ie ro (97) concluded t h a t c o b a l t p o ly c y th e m ia in r a t s was i n h i b i t e d by c y s t e i n e , i n h i b i t e d l e s s by h i s t i d i n e and n o t i n h i b i t e d by m e th io n in e . These w ork ers were ab le to o b t a i n some d e g re e o f p o ly c y th e m ia by th e a d m i n i s t r a t i o n o f th e c o b a l t h i e t i d i n e complex b u t n o t w ith th e c o b n l t - c y s t e i n e complex. E ly e t a l . (45) d em o n strated th e a b i l i t y o f i n j e c t e d m ethionine t o p re v e n t th e to x ic synptoms due to e x c e s s iv e c o b a lt i n t a k e in c a t t l e . - 9 O rten and O rten (92) found t h a t r a t s fe d a r a t i o n low i n p r o t e i n b e crne nolycytherr.ic a t th e sane r a t e an d to th e same e x t e n t a s d id r a t s on a sto ck r a t i o n . E ly e t a l . ( ^ t) were u n ab le t o reduce th e t o x i c i t y o f c o b a lt fe d t o d a i r y c a lv e s by supplem enting th e g r a i n m ix ture w ith c a s e i n , more th an d o u b lin g th e n r o t e i n c o n te n t of the g r a i n r a t i o n . Anderson et. a l . ( l ) we re able to in c r e a e e th e r a t e o f p o ly c y th e m ia development and i n t e n s i t y o f t o x i c i t y a s w e ll a s to d e p r e s s growth due to c o b a lt a d m i n i s t r a t i o n by th e a d d i t i o n o f mono-sodium n h o s n h a te to a m ilk d i e t , lo w e rin g th e calciu m /p h o sp h o ru s r a t i o o f th e m ilk to 0 . 6 . Orton e t a l . (102) d em o n strated th e s t a b i l i z i n g in f lu e n c e o f manganese on the i n c r e a s e d hemoglobin, e r y t h r o c y t e c o u n t, c e l l volume and b lo o d volume v a lu e s c h a r a c t e r i s t i c o f c o b a lt p o ly c y th e m ia . The same w o rk ers (100) found t h a t a d i e t c o n s i s t i n g o f whole m ilk supplem ented w ith adequate amounts o f iro n and coprxii w ith c o b a lt added would support p o ly c y th e m ia in r a t s f o r n e a r l y a y e a r and t h a t upon removal o f t h e c o b a lt from t h e d i e t th e hem oglobin, re d blo od c e l l volume, and e r y t h r o c y t e count slo w ly r e t u r n e d to norm al. Beard and Andes (9 ) o b ta in e d an i n c r e a s e i n r e d b lo o d c e l l co u n t w ith o u t i n c r e a s e in hemoglobin c o n c e n t r a t io n when r a t s were f e d m ilk f o r t i f i e d w ith i r o n and c o b a lt. A d d itio n o f v ita m in D to th e r a t i o n allo w ed th e hem oglobin concen­ t r a t i o n to in c r e a s e as w e l l . I t h a s been e s t a b l i s h e d t h a t c o b a l t p o ly c y th e m ia i s accompanied by an in c r e a s e in blood volume in r a t s ( lO l, 119) and dogs (ifO) . In each case th e in c r e a s e d blood volume was acco u n ted f o r by th e I n c r e a s e in volxrae of c e l l s . S ta n le y et, a l . ( lO l) c h a r a c t e r i z e d th e e r y t h r o c y t e s o f p o ly cy th em ic r a t s as b ein g h-1 p e r c e n t g r e a t e r i n volume th a n norm al, alm ost e n t i r e l y due to in c r e a s e d t h i c k n e s s o f th e c e l l . Th^rfound e s s e n t i a l l y th e same -1 0 - hemoglobin c o n t e n t i n b o t h no rm a l and p o l y c y t h e m i c c e l l s . V a l e r i c (132) s t u d i e d th e m o r p h o l o g i c a l c h a n g e s b r o u g h t a b o u t by c o b a l t poly c y th e m in i n g u in e a p i g s . he found mild p o i k i i o c y t o s i s , a te n d en c y t o a r c r e d u c t i o n i n e r y t h r o c y t e d i a m e t e r , a u g m e n t a t i o n o f lym ph oc y te s a f t e r p r o l o n g e d t e a t ment, and mo de ra te i n c r e a s e i n number o f p l a t e l e t s . V a l e r i o (1.33) d e t e r m i n e d t h a t c o b a l t t r e a t m e n t o f g u i n e a p i g s pr odu ce d hyperemia and c o n g e s t i o n o f bone marrow, l i v e r , s p l e e n , kid ne y a n a lung i n a d d i t i o n t o a p o l y c y t h e m i a o f m e d u l l a r y o r i g i n and r e t i c u l o h i s t i o c y t i c a c t i v i t y w i t h f o r m a t i o n o f he rrocytes and iuhagocyt.es. Observations of m i c r o s c o p i c a l l y v i s i b l e d e g e n e r a t i o n i n the l i v e r , k i d n e y s , p a n c r e a s , and myocardium and i n f l a m m a t i o n i n t h e k i d n e y s were made. Davis e t a l . (42) ob­ s e r v e d e a r l y irarrov# h y p e r p l a s i a and a n i n c r e a s e i n e x t r a m e d u l l a r ; , e r y t h r o p o i ­ e t i c masses i n th e s p l e e n , l i v e r , cyt.lnjir.ic by c o b a l t etiminis Lrs t i o n . k id n e y s and a d r e n a l s in duc ks made p e l ; - C o n ti n u e d i n j e c t i o n o f c o b a l t o u s c h l o r i d e produ ce d e v i d e n c e o f marrow h y p o p l a s i a , d i s a p p e a r a n c e of e x t r a m o d u l l a r y e r y t h r o p o i e t i c f o c i , and a lo w e re d p h a g o c y t i c a c t i v i t y of the r e t i c u l o ­ e n d o t h e l i a l e l e m e n t s o f the s p l e e n and l i v e r . Die l i t e r a t u r e c o n c e r n i n g t h e e f f e c t o f s p l e n e c t o m y on tho d e v e l o p ­ ment o f p o l y c y t h e m i a due to c o b a l t a d m i n i s t r a t i o n i s c o n t r o v e r s i a l . Cornil e t a ]. . (32) o b s e r v e d p a r e n c h y m a to u s d e g e n e r a t i o n o f th e l i v e r s o f g u i n e a p i g s re p e a te d ly in je c te d with sm all doses of c o b a lt a c e t a t e . was r a r e i n t e n s e i n s p l e n e c t o m i s o d a n i m a l s . The d e g e n e r a t i o n Porwald (1 4) r e p o r t e d t h a t s p l e n e c t o m v mode c o b a l t i n e f f e c t i v e a s a n a g e n t c a u s i n g p o l y c y t h e m i a i n r a t s b u t O r t e n ( ? £ ) found o n l y a d e l a y i n trie a c t i o n o f c o b a l t a t t r i b u t a b l e t o spenectoir.y. P a r r on ( ? ) found no change i n t h e a c t i o n o f c o b a l t on r a b b i t s due t o s p l e n e c t o m y . B u cc ie ro end O rten (23) c o u ld f i n d no evid en ce o f a d e c r e a s e in th e oxygen c a r r y i n g c a p a c i ty o f th e b lo o d of c o b a l t t r e a t e d p o ly c y th e m ic r a t s * The raethemoglobin c o n te n t o f th e b lo o d d id n o t v e ry s i g n i f i c a n t l y from th e c o n c e n t r a t i o n found i n th e blood o f c o n t r o l a n im a ls . Dorrance et, al_. (4-3) observ ed t h a t r a t s made p o ly c y th e m ic by c o b a lt a d m i n i s t r a t i o n have an in c r e a s e d work p erfo rm an ce u n d e r c o n d i t i o n s o f a n o x ia . Im m ediately a f t e r w ith d raw al o f th e c o b a lt a d m i n i s t r a t i o n and p r i o r to th*> drop in hemoglobin c o n c e n t r a t i o n o f th e blo od a s te a d y improvement i n work n e r formsnce was n o te d . B arron (?) showed t h a t c o b a lt added to p o ly cy th em ic blood in a Warburg a p p a r a t u s d e c r e a s e d r e s p i r a t i o n r a t e o f th e r e d b lo od c e l l s which may accou nt f o r th e r e s u l t s o b ta in e d by Dorrance a t al,. With­ draw al o f th e c o b a l t a d m i n i s t r a t i o n allo w ed th e c o b a l t c o n te n t o f th e c i r ­ c u l a t i n g b lo o d to f a l l n e a r l y to z e r o . Warren et, a l . ( 136) d i s a g r e e w ith Barron in r e g a r d to th e d e c r e a s e in r e s p i r a t i o n o f c e l l s i n v i t r o due to sm all c o n c e n t r a t i o n s o f c o b a l t . Mechanism o f C obalt P o ly cy th em ia V ario u s t h e o r i e s have been advanced in a t t e n p t s t o e x p l a i n th e p ro ­ d u c tio n o f c o b a l t p o ly c y th e m ia . Mascherpa ( 86) e x p la in e d th e a c t i o n o f c o b a l t a s a s t i m u l a t i o n o f bone marrow t o produce r e d and w h ite blood c e lls. He b e l i e v e d t h a t c o b a lt s t i m u l a t e s th e ly m p h o p o ie tic o rg an s and the sp le e n s in c e th e p r o d u c t io n o f w h ite c e l l s was a l s o i n c r e a s e d . Orton (96) concluded t h a t th e h e m a to o o ie tic a c t i v i t y o f c o b a lt i s a r e s u l t o f an i n c r e a s e in t h e r a t e o f f o rm a tio n o f hemoglobin and e r y t h r o c y t e s a s - 12 opposed to a p a s s i v e a c c u m u la tio n o f red c e l l s a s a restfLt o f a d im in ish e d ra te of c e ll d e s tru c tio n . The p o s s i b l e a c t i o n o f c o b a l t to i n h i b i t enzymes which c o n t a i n s u lf h y d r y l grouos h a s been p o s t u l a t e d a s a mechanism in p o ly c y th e m ia p r o d u c tio n (7, 57)* T h is a c t i o n would a l t e r th e o x i d a t i v e mechanisms a t th e c e l l u l a r l e v e l , l e a d ­ ing t o t h e i n h i b i t i o n of r e s p i r a t i o n o f th e young c e l l s and c a u s in g them to be thrown in to c i r c u l a t i o n ( 7 ) . O rten and B ucciero (97) a tte m p te d to ex- n l a i n th e a c t i o n o f c o b a lt on s u lf h y d r y l comoounds a s le a d i n g t o c e l l u l a r a n o x ia and, i n t u r n , l e a d i n g t o a com pensatory p o ly c y th e m ia . D avis (**l) a t t r i b u t e d t h e nolycythemi.a t o th e p' y s i o l o g i c a l e f f o r t to overcome l o c a l hypoxia pro du ced by c o b a l t . Schubmehl e_t a l . ( l l 3 ) concluded t h a t t h e chem ical d e p r e s s io n o f bone marrow r e s p i r a t i o n and g l y c o l y s i s was not th e mechamism in v o lv e d in c o b a lt p o ly c y th e m ia . These w o rk ers r u l e d out th e involvem ent o f n e u r a l mechanisms and h i s t o l o g i c a l changes in th e marrow blood v e s s e l s a s f a c t o r s . A r e c e n t c o n t r i b u t i o n may shed some l i g h t on th e mechanism o f c o b a lt p o ly c y th e m ia . B e r l i n (13) h a s shown by th e u s e o f a t r a c e r te c h n iq u e t h a t i n j e c t e d c o b a l t r a n i d l y r e a c h e s a s a t u r a t i o n l e v e l in th e l i v e r , k id n ey , and Ixinga o f r a t s , and t h a t n ro lo n g e d a d m i n i s t r a t i o n te n d s t o a slow i n ­ c r e a s e in the c o b a l t c o n te n t o f r e d b lo o d c e l l s , s p le e n , and bone marrow. C obalt T oleran ce in Ruminants S e v e ra l r e n o r t s have i n d i c a t e d the p o s s i b i l i t y o f r e a c h in g a to x ic l e v e l o f c o b a l t i n t a k e f o r c a t t l e and sheep. J o s l a n d (66) r e p o r te d t h e development o f a p o ly c y th e m ia i n one o f f o u r ewes dren ch ed d a i l y f o r seven months w ith one m illig ra m o f c o b a l t a s c o b n lto u s s u l f a t e p e r ?00 grams o f - 13 body w e i ^ i t . Two ewes became m ild ly anomic w h ile one was u n a f f e c t e d . S ly e t a l . (45) concluded t h a t o r a l a d m i n i s t r a t i o n o f e x c e s s iv e amounts o f co­ b a l t d e p re s s e d a p p e t i t e an d t h a t e x c e s s iv e i n t r a v e n o u s a d m i n i s t r a t i o n s o f c o b a lt s a l t s p rod uced r a p i d r e s p i r a t i o n , i n c o o r d i n a t i o n , l a c r i m a t i o n , s a l i v a t i o n , d e f e c a t i o n , and u r i n e le a k a g e w i t h i n f i v e m in u te s. These in ­ v e s t i g a t o r s (4£) concluded t h a t th e t o x i c i t y o f c o b a l t fe d 8s th e s u l f a t e , c h l o r i d e , o r c a rb o n a te was o f th e same i n t e n s i t y f o r c a l v e s . T o leran ce o f c a t t l e f o r c o b a l t h a s been e s t a b l i s h e d a t th e l e v e l o f 50 m illig r a m s p e r anim al p e r day ( 52) , ^0 m illig ra m s d a i l y p e r 100 pounds body w eig ht f o r c a lv e s (^+6), and 50 m illig ra m s d a i l y p e r 100 pounds body w eight f o r "grow­ in g d a i r y a n im a ls " (70)» S u lfa g u a n id in e M arsh all et. a l . ( 8 l ) f i r s t su g g ested t h e use o f s u lf a g u a n id in e as a c h e m o th e ra p e u tic a g e n t to combat i n t e s t i n a l i n f e c t i o n s . These w orkers d e s c r i b e d th e compound a s b e in g f a i r l y s o lu b le in w a te r b u t p o o r ly absorbed from th e i n t e s t i n a l t r a c t . Black et. al_. ( 15 ) were f i r s t to r e p o r t t h a t in ­ c l u s i o n o f 0 . 5 p e r c e n t s u lf a g u a n id in e in a s y n t h e t i c r a t i o n was s u f f i c i e n t to cause r e d u c t i o n in growth r a t e o f r a t s . C on firm atio n o f t h i s o b s e r v a tio n was made b y s e v e ra l w o rk e rs (33• 3 5» 79, 80, 8^, 85, 1 1 8 ). Black e t a l . ( 1 5 ) were a b le to prod uce a growth re sp o n s e vdien l i v e r e x t r a c t o r p-am inobenzoic a c id was i n c lu d e d in th e d i e t w ith s u lf a g u a n id i n e . The re sp o n s e to t h e s e two supplem ents d i f f e r e d i n t h a t th e re sp o n s e o f l i v e r e x t r a c t was immediate w h ile t h a t o f p-am ino b enzoic a c id was d e la y e d . These w o rkers i n t e r p r e t e d t h e i r r e s u l t s a s i n d i c a t i n g t h a t t h e p-am inobenzoic a c id overcame th e i n h i b i t i o n o f b a c t e r i a l s y n th e s i s caused by s u lf a g u a n id in e w hile l i v e r e x t r a c t f u r n i s h e d n u t r i t i v e p r i n c i p l e s o r d i n a r i l y e l a b o r a t e d by the b a c t e r i a o f th e a li m e n t a r y c a n a l . MacKenzie a t a l . (Po) r e p o r t e d hyperemi end h y p e rtro o h y o f t h r y r o i d g la n d s due to i n c l u s i o n o f one to two p e r c e n t s u lf e g u a n id in e i n the r a t r a t i o n in a d d i t i o n to co n firm in g th e r e s u l t s o f Black et, a l . V ario u s v ita m in d e f i c i e n c i e s have been induced by t h e a d m i n i s t r a t i o n o f sulfonam ide d e r i v a t i v e s to e x p e rim e n ta l a n im a ls . I t i s n o t th e p u rp o se o f t h i s review to cover t h i s phase o f t h e a c t i o n o f th e s u l f a d r u g s . For t h i s coverage th e r e a d e r i s r e f e r r e d to ad eq u ate re v ie w s (3* 3^» lb 4 ) • Gant a t a ^ . ( 5 l ) s tu d i e d th e i n t e s t i n a l f l o r a o f r a tB r e c e i v i n g a d i e t c o n ta in i n g s u lf e g u a n id in e and found t h a t the number o f organism s remained c o n s ta n t in the cecum but t h a t c o lifo r m organism s d e c r e a s e d i n number w h ile th e e n t e r o c o c c i i n c r e a s e d . Kirby a t al,. ( 73) d e m o n stra te d th e same s h i f t o f organism s i n f e c e s from human b e in g s t r e a t e d w i t h s u l f a s u x i d i n e . T h e ir r e s u l t s i n d i c a t e d t h a t E s c h e r i c h i a c o l i were th e organism s r e s p o n s i b l e f o r some v ita m in s y n th e s e s . The d e c re a se i n f e c a l e x c r e t i o n o f th ia m in e a s the r e s u l t o f s u l f a t h a l i d i n e a d m i n i s t r a t i o n to d a i r y cows h a s been r e p o r t e d (125)• T his would i n d i c a t e some change in the q u a l i t y o r q u a n t i t y o f th e f l o i o o f the a l im e n t a r y c a n a l . The mode o f a c t i o n o f sulfonam ide compounds h a s n o t been a c o n tro ­ v e r s i a l s u b je c t a lth o u g h , to d a t e , i t h a s n o t been d em o n strated e x p e r im e n ta lly in i t s e n t i r e t y . Woods and E l i d e s (lb-3) p o s t u l a t e d and Woods (lb-O) expanded a p o s s i b l e mechanism o f sulfonam ide compounds. These w o rk ers found p-amino­ b en zo ic a c i d to be a p o w erfu l en tiB ulfo nam id e agent a s w e ll a s an e s s e n t i a l n u t r i e n t e i t h e p s y n th e s iz e d by b a c t e r i a l c e l l s or o b ta in e d from th e en v iro n ­ ment o f th e c e l l s , a c c o r d in g to the ty p e o f b a c t e r i a . T h is t h e o r y a t t r i b u t e s th e c o m p e titio n between su lfo n am id es and. o-am in ob enzo ic a c i d to t h e i r common chem ical s t r u c t u r e s . The a n i l i d e n o r t i o n o f t h e i r m o lecu les i s common to b o th . The sulfonam ide d e r i v a t i v e competes f o r th e p a r t i c u l a r c e l l r e a c t i o n which r e q u i r e s p-am in ob en zo ic a c i d and which t a k e s p l a c e th ro u g h th e a n i l i d e p o r t i o n o f t h e m o lecu le. When th e sulfonam ide h a s r e a c t e d , t h e c h a in o f e v e n ts w i t h i n th e c e l l i s b ro k en sin c e the sulfonam ide lacks, th e rem ain in g c o n f i g u r a ti o n r e o u i r e d ana th e c e l l e i t h e r d i e s or i t s normal mebabolism i s m o d ified . This th e o r y does not accou nt f o r th e a n t i b a c t e r i a l a c t i o n o f a l l sulfonam ide d e r i v a t i v e s (53) b u t s u lf a g u a n id in e i s not one o f th e excep­ tio n s. Woods ( l ^ l ) h a s rev iew ed th e l i t e r a t u r e and comnounded a l i s t o f organism s i n c l u d i r g b a c t e r i a , f u n g i, and y e a s t s f o r which p-am in ob enzo ic a c i d i s an e s s e n t i a l growth f a c t o r . Many o t h e r o rganism s w hich do n ot r e ­ q u ire p -am in ob en zo ic a c i d have been found to be a b le t o s y n th e s i z e i t o r m a t e r i a l s w ith th e same b i o l o g i c a l a c t i v i t y . Wood (139) deduced t h a t s in c e th e equim olar r a t i o betw een th e c o n c e n t r a t i o n s o f p -am in ob enzoic a c i d and sulfonam ide d e r i v a t i v e s rem ain c o n s ta n t t h a t t h e s e two s u b sta n c e s compete f o r an enzyme. Tamura ( l 2 k ) showed t h a t th e a n tis u lf o n a m id e e f f e c t o f p - nminobenzoic a c i d o c c u rre d w ith a l l m icroorganism s t e s t e d w ith th e e x c e p tio n B n c t• t u l a r e n s e . Woods ( l ^ ? ) r e c e n t l y review ed th e b io c h e m ic a l s i g n i f i c a n c e o f p a n in o b en zo ic a c id -s u lfo n a m id e c o m p e titio n and the e f f e c t s o f t h i s com peti­ t i o n on th e f a c t o r s e i t h e r e l a b o r a t e d o r r e o u i r e d by m icro o rg an ism s. EXPERIMENTAL PROCEDURE Seventeen c a lv e s whose a g e s ran g ed from 12 t o 191 d ays when s t a r t e d on th e experim ent r e p r e s e n t i n g th e H o l a t e i n , G uernsey, J e r s e y , A y rs h ire , and Brown Swiss b r e e d s and e i g h t y e a r l i n g h e i f e r s o f th e H o l s t e i n b r e e d in th e M ichigan S t a t e C ollege Experim ent S t a t i o n d a i r y h erd were u se d in th is in v e stig a tio n . A ll a n im a ls hod been u se d f o r one o r more e x p erim en ts p r e v i o u s to b e in g a s s ig n e d to t h i s w ork. C r i t e r i a f o r th e s e l e c t i o n o f th e s e an im als were normal h e a l t h and a o p earan ce, a p p e t i t e f o r th e concen­ t r a t e m ix tu re , and normal b lo od p i c t u r e . The p e r t i n e n t d a ta r e g a r d in g th e s e an im als and t h e i r assign m ent to g r o u p s - i s p r e s e n t e d in Table 1 . C alves were s t a r t e d on the experim ent a t a l l tim e s o f y e a r a s th ey became a v a i l a b l e . The c a l v e s were m a in ta in e d in i n d i v i d u a l s o l i d b oard p e n s bedded w ith wood s h a v in g s . A ll c a lv e s had a c c e s s to good grade a l f a l f a mixed hay f o r ad l i b i t u m consum ption. Herd m ilk was fed a t th e r o t e o f 5 pounds d i l u t e d w ith w a t e r tw ice d o i l y to a l l c a l v e s except t h r e e anim als which were u sed on a p r e l i m i n a r y exp erim en t and were more advanced i n age when s t a r t e d on the e x p e rim e n t. A g r a i n m ix tu re c o n s i s t i n g o f bO p a r t s ground y ello w co rn, 40 p a r t s ground o a t s , 26 p a r t s l i n s e e d o i l meal and 1 p a r t s a l t was fe d t o a l l a n im a ls w h ile d a t a were b e in g ta k e n . The c o b a lt c o n te n t o f th e hays u se d i n t h i s ex p erim en t ranged from 0 .0 3 t o 0 .1 8 p o r t s p e r m i l l i o n , o f th e g r o in m ix tu re from 0*04 to 0*18 p a r t s p e r m i l l i o n , and o f th e m ilk from 0 .0 0 2 t o 0 .0 0 7 p a r t s p e r m i l l i o n . The y e a r lin g h e i f e r s were a l l Btarted on th e experim ent a t one tim e. Their r a tio n was composed o f the same type o f hay and the same concentrate mixture as that fed to th e c a lv e s . These h eife rB were kept in stan ch ion s and Were fed in mangers having t ig h t p a r titio n s * A p r e l i m i n a r y experim ent was s o t up u s i n g a n im a ls C680, C68l, and C68h. C obaltous s u l f a t e , h e p t a - h y d r a t e , was mixed i n t o th e c o n c e n tr a te m ixture a t the r a t e o f 0 . 5 gram p e r pound o f m ix tu re and fed a t the d a i l y r a t e o f 1 pound p e r 105 pounds body w e ig h t. T h is su p p le m e n ta tio n f u r n is h e d 1 m illig ra m c o b a lt p e r pound body w eight p e r day . I t was un­ f o r t u n a t e t h a t th e anim al which was d e s ig n a te d to be th e c o n t r o l in t h i s experim ent d i e d soon a f t e r c o b a l t a d m i n i s t r a t i o n was s t a r t e d on th e o t h e r t h r e e a n im a ls . In th e l i g h t o f t h i s development i t was d e c id e d t o d e l e t e c o b a l t from t h e r a t i o n o f anim al C681 a f t e r 136 days o f s u p p le m e n ta tio n . The c o n t r o l f o r t h i s group c o n s i s t e d o f th e u e r i o d b e f o r e su p p lem en tatio n f o r each anim al and th e p e r i o d subsequent to s u p p le m e n ta tio n i n th e case o f c a l f C681. Four groups o f c a l v e s were s e t up on th e b a s i s o f r e s u l t s o b ta in e d w ith the p r e l im i n a r y g ro u p . c e iv e d no supolem ent. The c o n t r o l g rou p, d e s ig n a te d Group 1, r e ­ Group 2 was supplem ented to t h e e x te n t o f 1 m i l l i ­ gram c o b a l t d a i l y p e r pound body w eight by i n c o r p o r a t i n g 1 gram c o b a lto u s s u l f a t e , h e p t a - h y d r a t e , i n each pound o f c o n c e n t r a te m ixtu re and fe e d in g th e supplem ented m ix tu re a t t h e r a t e o f 1 pound d a i l y f o r each 210 potmds body w e i g h t . A f te r 1^ weeks o f c o b a l t s u p p le m e n ta tio n animal 07^1 was supplem ented w ith 1 m illig ra m i r o n d a i l y p e r pound body w eig h t s u p p lie d i n t h e form o f f e r r i c c i t r a t e , C. P . Feeding s c h e d u le s were r e v i s e d weekly and th e c o n c e n tr a te r a t i o n was mixed and bogged f o r t h a t p e r i o d o f tim e . The s u p p le m e n ta tio n f o r Group 3 d i f f e r e d from t h a t o f Group 2 by the a d d i­ t i o n o f s u lf a g u a n id in e t o th e e x te n t o f 2 p er c e n t o f t h e e s ti m a t e d d ry m a tte r o f t h e r a t i o n mixed i n t o th e c o b a l t - c o n c e n t r a t e m ix tu re f o r 70, 102, - 18 end 115 d ays, r e s p e c t iv e ly , o f the time th ese anim als were on the ex p eri­ ment* Subsequent to th ese p erio d s the co b a lt supplement was con tin u ed . The two a n im a ls in Group h were supplemented f o r p e r i o d s of 39 and 1?5 days, r e s p e c t i v e l y , w ith s u lf a g u a n i d i n e alo n e a t th e r a t e o f 2 p e r c e n t o f t h e e s tim a te d d ry m a tte r in ta k e * F o llow in g th e s e p e r i o d s o f time th e a n im a ls were supplem ented w ith c o b a l t a t th e sane r a t e as were the an im als i n Group 2. A d d itio n a l uneupolem ented c o n c e n tr a te m ixture was in c lu d e d i n t h e d a i l y con­ c e n t r a t e r a t i o n o f any anim al in Groups 1 to h, i n c l u s i v e , which had not g ain ed w eight th e p r e v i o u s week. Groups 5 and 6 were composed o f y e a r l i n g h e i f e r s . Group 5 Wfts th e c o n t r o l f o r Group 6 which was supplemented w ith c o b a l t a t th e r a t e o f 0*5 m illig ra m s d a i l y p e r pound body w eight- The supplement was s u p p lie d by ?5 grems c o b a l t o u s s u l f a t e , h e p t a - h y d r a t e , mixed i n t o 100 pounds o f c o n c e n tr a t e m ixture a n d fe d a t th e r a t e o f 1 pound d a i l y p e r 105 pounds body w e ig h t. A f te r 8h days on exo erim en t th e c o n c e n t r a t e m ix tu re was supplemented w ith 50 grams c o b a lt o u s s u l f a t e , h e p t a - h y d r a t e , p e r 100 pounds o f c o n c e n tr a te m ixtu re f e d a t *he r a t e o f 0 .7 5 uound p e r day p e r 105 pounds body w e ig h t. This l a t t e r su p p lem en tatio n "/as d a i l y p e r pound body w e i g h t . s u f f i c i e n t to f u r n i s h 0*75 m illig ra m c o b a l t Group 5 r e c e iv e d unsupplem ented c o n c e n tr a te m ixture a t th e same r a t e s . Three t e s t tu b e s o f j u g u l a r b l o c ’ wore taken w eekly, one c o n ta in in g l i t h i u m c i t r a t e as a n t i c o e g u l a n t , one c o n ta i n i n g p o ta ssiu m o x a l a t e , and one w ith no a n t i c o a g u l a n t . The c i t r a t e d b lo o d was u s e d f o r d e te r m in a tio n o f r e d b loo d c e l l count by th e method o f Wlntrobe (13P, p . 2 l l ) , r e d blood c e l l volume ( h e m a to c rit) by the method o f Wintrobe (138 p . 201), hemoglobin by ^ - 19 th e method, o f Sanford e t a l . (1 0 9 ), plasm a calcium by th e method o f C lark end C o llip ( 2 6 ), plasm a in o r g a n ic p h osph o ru s by B r ig g s ' m o d i f i c a t i o n (19) o f th e B e ll and Doisy method (1 2 ), and p lasm a magnesium by a m o d i f i c a ti o n d e v is e d by Duncan e_t_ al_. (44) o f th e B rig g s method ( 2 0 ) . ac id wub p lasm a. Plasm a a s c o r b i c d ete rm in e d by th e method o f M indlin and B u t l e r (90) in o x a la te d T o ta l serum p r o t e i n s and serum albumin were d ete rm in e d by a m icro - K je ld a h l t e c h n i q u e . G lo b u lin s e p a r a t i o n was accom plished by K i n g s l e y 's m o d if ic a tio n (72) o f Howe's method ( 6 l ) . C a l c u la t io n o f mean c o r p u s c u la r hem oglobin, mepn c o r p u s c u l a r volume, and mean c o r p u s c u la r hem oglobin c o n c e n t r a t io n v a l u e s were made from the e x p e rim e n ta l d a t a . Each anim al was weighed w eekly w h ile on the e x p e rim e n t. Response to su p p le m e n ta tio n was e v a l u a t e d on th e b a s i s o f body w eight change and b lo o d a n a l y s e s . T able 1 Composition o f E x p e rim en tal Groups Group Animal No. Breed ...... ................. ... Sex At s t a r t (days) Supol. S t a r t e d (dBys) P r e li m i n a r y C630 C68l C68U- Guernsey Guernsey H o l s te i n M M M 191 175 l6 l 210 195 181 1 . C o ntro l C718 C719 C724 C742 0749 Brown Swiss A y rsh ire Jersey H o l s te in Guernsey M M M M M 45 18 32 33 75 — — — - C715 0739 0741 0744 Jersey H o l s te i n H o lste in H o l s te i n M 32 35 33 12 39 53 36 29 81 32 37 88 50 47 46 32 84 57 431 383 352 288 455 363 354 345 2. Cobelt M M M 3» C obalt p l u s S u lfa g u a n id in e — Cobalt H o l s te i n C711 H o l s te i n 0725 H o l s te i n C730 M M 4 . S u lfa g u a n id in e — C obalt 0714 H o lste in H o ls te in C? 26 P M M 5. Y e a r lin g H e i f e r s —■Control H o l s te i n A73 H o lste in A75 A78 H o lste in A80 H o l s te i n P P 371 323 292 228 6 . Y e a r l i n g H e i f e r s —■Cobalt A72 H o l s te i n A76 H o l s te i n H o l s te i n A77 H o l s te i n A79 P P P P 395 303 294 285 P F Blond P i c t u r e The d a t a from d e t e r m in a tio n s perform ed on th e b lo o d from a n im a ls used i n the p r e l im i n a r y t r i a l a re p r e s e n t e d in Appendix T a b le s 1, 2, and 3 and th o se p e r t a i n i n g to whole b lo o d i n F ig u r e s 1, 2, and 3* ®iese th r e e a n im a ls reach ed th e h e i g h t o f t h e i r i n c r e a s e s i n r o d bloo d c e l l co un ts a p p ro x im a te ly 15 weeks a f t e r c o b a l t su p o le m e n ta tio n was s t a r t e d . Animal number C68l m a in ta in e d th e h ig h re d blo nd c e l l count u n t i l ab ou t 1+ weeks a f t e r su p p lem en tatio n was suspended -jrherevs the co u n ts spon taneou s­ l y reduced i n i n t e n s i t y in th e o t h e r two c a s e s a t about 55 weeks o f a g e . Hemoglobin and r e d b lo o d c e l l volume v a l u e s e s s e n t i a l l y p a r a l l e l e d th o se o f th e re d blood c e l l c o u n ts . L i t t l e change was n o te d i n the mean c o r p u s c u la r volume, mean c o r p u s c u la r hem oglobin, and mean c o r p u s c u la r hemoglobin c o n c e n t r a ti o n v a l u e s f o r th e d u r a t io n o f th e t r i a l . Peak ex­ p e r im e n ta l v a l u e s o b ta in e d were l 6 . ? 6 , l6*31» flud 18.10 m i l l i o n s r e d b lo o d c e l l s p e r c u b ic m i l l i m e t e r , 1 9 .0 0 , 1 9 .2 0 , and 18.80 grams p e r cent hemo­ g lo b in , and 50*0 , 55*5. a ud hS»5 p e r c e n t r e d b lo o d c e l l volume, r e s p e c t i v e ­ ly . These v a l u e s r e p r e s e n t i n c r e a s e s in e x c e s s o f 60 p e r c e n t as compared t o th o se o b ta in e d p r i o r to c o b a l t su p p le m e n ta tio n . The e x p e rim e n ta l v a l u e s f o r plasm a calcium , p lasm a in o r g a n ic p ho sph orus, plasm a magnesium, and plasm a a s c o r b ic a c id were r e l a t i v e l y c o n s ta n t and in no way fo llo w e d th e p a t t e r n o f v a l u e s s e t by th e measurements co n c e rn in g th e r e d blood c o l l s . A l l t h r e e o f th e an im als g ain ed w eight i n e x c e s s o f 1 pound p e r day d u rin g the p e r i o d o f e x p e r im e n ta tio n a s i s shown in Appendix Table 26. The d e t a i l e d d a t a o b ta in e d on Group 1, th e c o n t r o l group, a re p r e ­ sen te d in Appendix T a b le s 4 , 5 , 6 , 7, and 8 and th e d a ta o b ta in e d on the whole blood end serum o f t h i s group a r e p r e s e n t e d i n F ig u re s 4 , 5, 6. 7 , and 8 . Those o b ta in e d on Group 2, th e c o b a l t supplem ented group, a r e p r e s e n t e d i n Apoendix T a b le s 9* 10, 11, and 12 and i n F ig u r e s 9, 10, 11, and 12. Comparison o f th e r e s u l t s o b ta in e d w ith Groups 1 and 2 showed e s s e n t i a l l y no d i f f e r e n c e i n c o n c e n t r a t i o n s o f c o n s t i t u e n t s d eterm in e d on b lo o d plasm a a s w o ll a s i n c o n c e n t r a t i o n s o f t o t a l serum p r o t e i n s and serum album in. D i f f e r e n c e s between th e two grouos d id occur in the r e s u l t s o b ta in e d from d e t e r m in a tio n s made on th e e r y t h r o c y t e s and t h e i r c o n s t i t u e n t s . By th e time th e an im als in Group 2 had been supnlem ented w ith c o b a lt f o r 15 weeks t h e i r r e d b lo o d c e l l c o u n ts had i n c r e a s e d a p p ro x im a te ly 100 p e r cent over th o s e o f Group 1, to v a l u e s ra n g in g from 1 8 .9 5 to 21*95 m i l l i o n s r e d b lo o d c e l l s p e r c u b ic m i l l i m e t e r . The r e d b lo o d c e l l volume v a l u e s l e v e l e d o f f 3 to 5 weeks a f t e r th e p l a t e a u was r e a c h e d i n the r e d blood c e l l c o u n ts . High r e d b lo o d c e l l volume v a l u e s ran g ed from 4 6 .5 t o 58*5 p e r c e n t f o r th e fo u r an im als i n Group 2. The hemoglobin c o n c e n tr a t io n d i d n o t l e v e l o u t u n t i l 10 t o 15 weekB a f t e r th e l e v e l i n g o cc u rre d i n re d blood c e l l c o u n ts and in one c a s e , t h a t o f C744 a s shown i n F ig u re 12, th e anim al was n o t c o n tin u e d on the ex p erim en t a s u f f i c i e n t l e n g t h o f time f o r th e hemoglobin c o n c e n tr a t i o n t o r e a c h a p l a t e a u . The mean c o r p u s c u la r hemoglobin v a l u e s e s s e n t i a l l y p a r a l l e l e d th e mean c o r p u s c u la r volume v a l u e s b o th i n Grouu 1 and i n Group 2. However, "both d e c re a s e d i n Group 2 an im als u n t i l such a time a s th e red b lo n d c o l l c o u n t8 reach ed t h e i r p l a t e a u s and t h e r e a f t e r th e s e mean v a lu e s ten d ed to r e t u r n to t h e i r o r i g i n a l l e v e l , w hereas b o th th e s e v a lu e s rem ained r e l a ­ t i v e l y c o n s ta n t f o r a n im a ls in Group 1 . The mean c o r p u s c u la r hemoglobin c o n c e n tr a t io n rem ained e s s e n t i a l l y unchanged in b o th groups f o r th e dura­ t i o n o f th e e x p erim en t. T his c o n d i t i o n may bo d e s c r i b e d a s a m ic r o c y tic hypochromic po ly cy th em ia which changes t o a n o rm o cy tic normochromic p oly cy th em ia a f t e r th e r e d b lo o d c e l l count p l a t e a u i s r e a c h e d . Supple­ m en tatio n w ith i r o n i n th e r a t i o n o f animal C?hl a p p a r e n t l y speeded up th e r e t u r n o f th e re d b lo o d c e l l s iz e and re d b lo o d c e l l hemoglobin con­ t e n t to n orm al. The d e t a i l e d d a t a f o r th e an im als i n Group 3 a r e p r e s e n t e d i n Appendix T a b le s 1 3 , 1^, and 15 ®nd th e d a t a o b ta in e d from d e te r m in a tio n s c a r r i e d out on the whole b loo d and serum from th e se an im als a r e p r e s e n t e d in F ig u r e s 13, l h , and 15* The v a lu e s f o r p lasm a and serum c o n s t i t u e n t s which were determ in e d showed no m a t e r ia l change d u r in g th e p e r i o d o f ex p erim en ta­ t i o n a s was found f o r Groups 1 and 2. During th e p e r i o d o f c o b a lt and s u lf a g u a n id in e su p p lem en tatio n the a n im a ls in t h i s group showed g ra d u a l c o n c u rre n t i n c r e a s e s in r e d b lo o d c e l l c o u n ts , ro d bloo d c o l l volumes and hemoglobin c o n c e n t r a ti o n s f o r p e r i o d s v a r y in g from 3 t o 10 weeks. Red b lo o d c e l l co u n ts r a n g in g from 15*85 t o 16.70 m i l l i o n s p e r c u b ic m i l l i m e t e r were a t t a i n e d i n t h i s g roup. In two o f th e t h r e e c a s e s th e r e d b lo o d c e l l volumes and hemoglobin c o n c e n t r a t i o n s a t t a i n e d p a r a l l e l e d th o se re a c h e d in Group 2. Mean c o r p u s c u la r hemoglobin v a l u e s , mean c o r p u s c u la r volumes and mean c o r p u s c u la r hemoglobin c o n c e n tr a t i o n s r e n a in e d e s s e n t i a l l y unchanged d u r in g t h i g p e r i o d . In o n ly one o f th e 3 cag es wag c o b a l t su p p le m e n ta tio n alone a b le to su p p o rt th e h ig h re d bloo d c e l l c o u n ts , red blo od c e l l volumes, end hemoglobin c o n c e n t r a t i o n s which were a t t a i n e d w h ile s u l f a g u a n id in e was in c lu d e d in the supplem ent. The d e t a i l e d d a t8 f o r th e two a n im a ls in Group b- a r e p r e s e n t e d in Appendix T ab les 16 and 17 end d a t a o b ta in e d on whole blood and serum from th e s e an im als e r e p r e s e n t e d in F ig u r e s l 6 and 1 ?. D uring th e p e r i o d o f s u lf a g u a n id in e s u p p le m e n ta tio n th e v a r i o u s v a lu e s f o r blo od c o n s t i t u e n t s which were determ in e d d id n o t very s i g n i f i c i a n t l y from th o s e v a l u e s ob­ t a i n e d w i t h Group 1 w ith the e x c e p tio n o f totriL serum p r o t e i n c o n c e n tr a ­ t i o n in th e case o f anim al number C??6 which s u f f e r e d an a t t a c k o f pneumonia d tirin g t h i s p e r i o d . A f te r the s u lf a g u a n id in e su p p lem en tatio n had been r e p la c e d w ith c o b a l t s u p p le m e n ta tio n th e r e s u l t s f o r bo th an im als resem bled th o se o b ta in e d w ith Groxip 3 dxiring c o n c u r re n t c o b a lt end s u l f a g u an id in e s u p p le m e n ta tio n a lth o u g h one an im al, C726, v a r i e d in the i n t e n s i ­ ty o f th e e f f e c t s . I t may be n o te d a t t h i s p o i n t t h a t t h i s anim al was sh o u t 8 weeks o ld e r th an t h e o th e r an im a l, C?l4, when c o b a lt su p p lem en tatio n was s t a r t e d . The d e t a i l e d d a t a o b ta in e d from d e te r m in a tio n s perform ed on th e blood o f y e a r l i n g h e i f e r s i n Group 5 a re p r e s e n t e d in Appendix T a b le s 1 8 , 19, 20, and 21 and tho se from Group 6 in Appendix T a b le s 22, 23, 2h, and 25» Repre­ s e n t a t i v e a n im a ls from th e s e two groups were s e l e c t e d f o r g r a p h ic a l p r e s e n ­ t a t i o n o f d a t a i n F i g u r e s 18 and 19, r e s p e c t i v e l y . No c o n s i s t e n t d i f f e r e n c e co uld be n o ted i n t h e d a t a from th e s e two g roups o f animals* One animal in th e c o b a lt supplem ented group, A72, d id show ev iden ce o f some e l e v a t i o n s o f r e d b lo o d c e l l volume and hemoglobin c o n c e n t r a t i o n . - 25 - Growth The d e t a i l e d d a t a c o n c e rn in g body w eig ht changes o f the a n im a ls used in th e exp erim ent a re p r e s e n t e d in Appendix Table 26. Hxe to the v a r i a ­ t i o n s i n ag e, sex, end b r e e d o f anim als us9d com parisons o f th e body w e ig h ts were made w ith w e ig h ts i n t e r p o l a t e d from R a g s d a le ’ s s ta n d a r d (105)• The r e s u l t s o f th e s e com parisons f o r Groups 1, 2, 3* ®nd. b a r e p r e s e n te d in F ig u re 20 and f o r Groups 5 °ad 6 i n F ig u re 21. on th e s e f i g u r e s r e p r e s e n t supplement changes* B reaks in th e cu rv es D ata from which the f i g u r e s were c o n s tr u c t e d a r e p r e s e n te d in Appendix T a b le s 27 and 28, resp e c tiv e ly . Without e x c e p tio n th e a n im a ls in Group 1 e x h i b i t e d r a t e s o f growth which were s u p e r i o r to th ose o f the s ta n d a r d . 2, 3. a r ^ grew a s r a p i d l y as th e s ta n d a r d . No anim al i n Groups Animal number C?lb in Group b most n e a r l y approached t h i s accomplishm ent b u t i t s r a t e o f growth d e c lin e d d u r in g th e p e r i o d o f c o b a lt s u p p le m e n ta tio n . The a n im a ls i n Groups 5 ®ad 6 e x h i b i t e d v e r y n e a r l y th e same growth p i c t u r e w ith one e x c e p tio n , A80, which su rp a s se d the s ta n d a rd and was in th e unsupplem ented group (Group 5)• i - 26 - Table 2 Key t o A b b r e v ia tio n s Used, on F ig u re s RBCC Red bloo d c e l l co u n t— m i l l i o n s p e r c u b ic m i l l i m e t e r . RBCV Volume o f p ack ed re d blo od c e l l s — p e r c e n t . HB Hemoglobin— grams p e r c e n t • MOV Mean c o r p u s c u la r volume— c u b ic m icro n s. MCH Mean c o r p u s c u la r hemoglobin— micromicrograms p e r re d b lo o d c o l l . MCHC Mean c o r p u s c u la r hemoglobin c o n c e n t r a t io n —p e r c e n t . TSP T o ta l Berum u r o t e in s — grams p e r c e n t. S. ALB Serum albumin— grams p e r c e n t . BODY WT . Body w e ig h t—hundredw eight. Hemoglobin (grams p e r 1000 cubic c e n t im e t e r s o f b lo o d ) MCH*- -------------------------------------------------------------------------------------------------Red b lo n d c e l l count ( m i l l i o n s p e r cu b ic m i l l i m e t e r o f blond) Volume o f oacked red blo nd c e l l s (p er 1000 c u b ic c e n t i m e t e r s o f blood) MCV*=---------------------------------------------------------------------------------------------------------------Red b lo o d c o l l count ( m i l l i o n s p e r c u b ic m i l l i m e t e r o f blood) Hemoglobin (grams u e r 100 c u b ic c e n t i m e t e r s o f blood x 100) MCHC*= --------------------- ------------------------------------------------------------------Volume packed r e d c e l l s ( p e r 100 c u b ic c e n t i m e t e r s o f b lond) + (138, p . 222-223) — RBCC «X3V WT — R8CV — M C V ---------MCH — MCHC 50 A / 30 o 20 50 ,40 V 30 20 30 F ig u re i - 45 50 WEEKS OF AGE 55 60 Blond p i c t u r e cr.d grov.-th curve of T .in a l r.urfber C630 - - P r e lim in a r y Br oun A 65 70 ro ro *2 H O CD (i) R0CV - 9 b co -»> o ro M P- C) c* p>1 (X > rtt t* p* (Ti ►1 £ ‘.3 ‘ O § tv o (-•> -a »-*• n (0 o M ro CX> H* 1 I (5 l~* >-*• n* M J3 3» 3 O ► 1 & " 3J " (r sj MCV-jj? ’MCHC - % (O CD 4k MCH - GM x IO-B -R B C C --R B C V -M C V -M C H C HB M CH CM X .4 0 CD 30 35 4 0I W EEKS F ig u re 3* 45 50 55 O F fiSE Blood p i c t u r e and growth curve o f animal number C££4— P r e lim in a r y Group. 60 I -RBCC —TSP —RBCV —MCV -MCHC HB S-ALB BOW WT MCH 40 CM O X co 40 o 00 CO 30 - A \ //y/ " '''A '. a2 20 cc 30 WEEKS OF ASE B lood T sietn re nnd ~rov;th ;ro rv e o f r r . i —l C n S — Orocto 1- Tror/bor 50r 20 OJ -R B C C HB -T S P S .A L B BODY W T M CH •RBCV -M C V -M C H C x40 o 16 o tb x 12 o 20 40 A 30 »o ' 20 i- o o CO acr 20 W E E K S OF A&E RBCC TSP RBCV MCV M CHC — HB — S .ALB — BODY W T — M CH ,40 £216 o 40 i- 30 o &20 o ccO 30 W EEKS OF AGE ¥ BOOY WT RBCV - % ro ro o HR B C C - x K ^ /M M 3 - CW T o ) ____ M C V -u 3 o> H 8, TSP, 6 _ & A L B -& M % td O o rd nt* -* o MCH - G M x 10-12 GO -0 M C H C -% G» o o ro _______ O \ ' 2® w i \ p* Vi o‘1 2 ► 1 -1 o •{ 'i 0 s o' d C*4 •0 r-~M I I CP >1 O oP •R B C C ■TSP RBGV ■M CV tvj HB -& A L B ■BODY W T M CH •M C H C /) CO ?30 CW T x o - CD BODY WT CL 20 x 8 no ex. co cc 0 20 W EEKS 30 OF A&E ■RBCC •TSP -ROCV MCV ■MCHC 40 ^ S.ALB BOOT WT MCH 16. o 30 ox ^20 60r 24 50 20 - 40 to >- 3r 130 cr 62 20 ’ o cr 25 WEEKS OF ABE « . r r . I'.. r :iI —n i :.u n b e r J " I *- - 'm ' . r . -jf TIT RBCC -T S P R0CV N1CV MCHC ,4 0 HB S.ALB BOCJT WT MCH CM O X o S30 50 40 LD 30 i-2 20 W E E K S OF A CE 25 30 •RBCC ■TSP ■RBCV ■MCV •MCHC 40 ■H0 &ALB BODY WT ■MCH CM 60 50 40 V a I 20 RBCC-xO^/MM3 o ____________ HB, TSP, S.ALB-GM% MCH-GIYlxlO'^2 cp _________ CD_________ ro _________ 5R *-*• o 'J O Oc. ro -RBCC —TCP —RBCV -SALB —BOCV WT -M CH -WDJ 40 -MCHC ' .) . CO 6 S W E E K S OF ABE Figure l j . B io o i u l c e r e *.r.d growth c* o f m i r a l r.'j:::er 7711--Grr.': \ RBCV — :m^ c— 0OW W T mch CO 6 S 25 50 WSKSOFABE ■RBCC TSP -R8CV ■MCV M CHC lz HB &ALB BOW W T ■MCH \ ___ 60 50 40 CD 3 0 <0 12 or BODY o ! o 30 W EEKS OF AGE n'-’ th. C 3.T V 5 o f on in o i iro rb e r C7~'~--0ro"o ■RQCC TSP BGCV MCV MCHC & L 60 SALB 000V WT ■MCH L 24- 5 0 -2 2 0 CD 40 to 10 ^ 4 WEEKS OF fliE BODY W T - CWT. 9 - RBCC-xD rcv% , H B -G M H S P 6 & A IB - - V» M CV-U3 MCHC - % M C H - Q M x 10-12 Q »-*• r1 u M O n *o »*• o M O fPj O 5* r«* :r £ 1 0 ) It f1 0) £ •IJ* >1 O 0 r> < 7\ 1 I C«> M > ■3 — HB — BODY W T — MCH o o40 o I s CD CO V ID '3 0 o <0 4 45 50 55 W EEKS OF A&E 60 65 -- HB — BODY \A/T — M CH rvj o ,4 0 o s5 o CD ls r30 o I.* V ;\ 30 £12 45 50 W EEKS 1° • 55 OF A&E 60 p i c i’-ir .• ■,£ m i:r.al n u rb c r A??— &rouo 6. 65 STD, DP I '4 9 / 7 4 2 724 &P 744 7 25 STD 715 -730 W T. STO, BODY o GP 4 726 RAGSDALE “ STANDARD W T . 3 CW T. R A & S D A IE - STANOARO W T . - 48 - DISCUSSION In o r d e r to e s t a b l i s h th e e x i s t e n c e o f and to c h a r a c t e r i z e the p o ly c y ­ them ic c o n d i t i o n i n c a t t l e i t was n e c e s s a r y to a n a ly z e th e blood o f an im als i n w hich t h i s c o n d i t i o n e x i s t e d . D e te rm in a tio n o f th e number o f r e d b lo o d c e l l s a s w e ll a s t h e c h a r a c t e r i s t i c s o f them y i e l d e d i n s u f f i c i e n t evidence f o r th e c h a r a c t e r i z a t i o n . A n aly ses f o r n o n c e l l u l a r c o n s t i t u e n t s o f b lo o d y i e ld e d much added in f o r m a t i o n . The f a c t t h a t th e c o n c e n t r a t i o n s o f th e se n o n c e l l u l a r c o n s t i t u e n t s , e s p e c i a l l y t h a t o f th e serum p r o t e i n s , w ere n o t g r e a t l y a l t e r e d a s th e number o f r e d b lo o d c e l l s i n c r e a s e d e l i m i n a t e d con­ c e n t r a t i o n o f th e b lo o d a s a f a c t o r i n t h e t> red u ctio n o f th e p o ly c y th e m ia . The p r o b a b i l i t y o f growth u n d e r th e c o n d i t i o n s o f such extreme blood-volum e d e c r e a s e a s would be n e c e s s a r y t o produce t h e l a r g e re d bloo d c e l l counts which were e n c o u n te re d i n t h i s experim ent i s v e ry s l i g h t . The d a t a i n d i c a t e t h a t th e r a t i o o f r e d b lo o d c e l l count to concen­ t r a t i o n s o f n o n c e l l u l a r c o n s t i t u t e n t s i s g r e a t l y i n c r e a s e d i n t h i s p o ly c y ­ th e m ia . T h is i s in a c c o r d w i t h th e r e s u l t s o b te in e d on c o b a lt- p o ly c y th e m ic r a t s by O rten fit a l . ( lO l) and S ta n le y e t a l . (119) who found i n c r e a s e d b lo o d volume due e n t i r e l y to i n c r e a s e in t h e volume o f t h e t o t a l e r y t h r o c y t e mass* R e s u l t s o b ta in e d w ith dogs concur w ith t h i s co n cep t ( 4 o ) . A d e c r e a s e in th e mean c o r p u s c u la r volume was e n c o u n te re d in th e blo od o f th e a n im a ls i n Group 2. A s l i g h t d e c r e a s e in e r y t h r o c y t e d ia m e te r h a s been r e p o r t e d i n g u in e a p i g s made p o ly cy th em ic b y c o b a l t a d m i n i s t r a t i o n ( 132) . T h is d e c r e a s e would r e f l e c t a more pronounced d e c r e a s e i n e r y t h r o c y t e volume. Since th e mean c o r p u s c u la r hem oglobin d e c r e a s e d c o n c u r r e n t l y w ith th e mean c o r p u s c u la r volume t h e mechanism in v o lv e d can o n ly be p o s t u l a t e d . Knoop gi. £l_. (76) have shown t h a t t h e e f f e c t o f i r o n d e f i c i e n c y i n d a i r y c a lv e s i 8 a d e c l i n e i n th e hemoglobin c o n c e n t r a t i o n c o n c u r re n t w it h a l e s s e r d e c l i n e in red b lo o d c e l l c o u n t. i n Group I t i s p o s s i b l e t h a t th e an im als 2 d i d not r e c e i v e enough i r o n i n t h e i r r a t i o n to s a t i s f y th e i n ­ c r e a s e d re q u ire m e n t cau sed by development o f th e p o ly c y th e m ia . T h is , in t u r n , may have been r e s p o n s i b l e f o r t h e d e c r e a s e d e r y t h r o c y t e volume th u s com pensating f o r i n s u f f i c i e n t hemoglobin s y n t h e s i s . S upp lem en tation o f th e r a t i o n o f one c a l f , 07^1, w ith i r o n in th e form o f f e r r i c c i t r a t e a t a p o i n t i n the experim ent when th e e r y t h r o c y t e s were sm all seemed to h a s t e n t h e i r r e t u r n t o normal s i z e . A d m itted ly a s i n g l e case i s i n s u f f i c i e n t f o r draw ing c o n c lu s io n s b u t the r e s u l t s w ith t h i s one anim al do a c c e n tu a te the p l a u s i b i l i t y o f th e f o re g o in g h y p o t h e s i s . The s i m i l a r i t y between t h e b lo o d p i c t u r e s o f t h e a n im a ls i n Groups 3 and 1+ d u r in g th e p e r i o d of c o b a l t a d m i n i s t r a t i o n a p p e a r s to be s u f f i c i e n t b a s i s f o r d i s c u s s in g t h e s e two groups a s on e. S u lfe g u a n id in e supplementa­ t i o n , w h eth er a lo n e o r i n c o n ju n c tio n w ith c o b a l t supplement p r e v i o u s to c o b a l t s u p p le m e n ta tio n a lo n e , a p p a r e n tl y p r e d i s p o s e d th e po ly cy th em ia to be of a le s s e r sev erity . In a d d i t i o n t h e e r y t h r o c y t e volume and e r y t h r o c y te hemoglobin c o n te n t rem ained a t about th e same l e v e l f o r th e d u r a t io n o f th e exp erim en t which i s i n d i r e c t c o n t r a s t t o t h e lo w e rin g o f th e s e two v a lu e s in th e b lo o d o f th e a n im a ls i n Group 2. Hemoglobin c o n c e n t r a t i o n s f o r an im als i n Groups 3 and k were not as h ig h a t th e h e i g h t o f t h e p o ly c y th e m ia \ - 50 - fle th e y w e re i n the Group 2 anim als* The e f f e c t o f s u lf a g u a n id in e on th e m i c r o f l o r a o f th e rumen must "be c o n s id e re d . Sulfonamide d e r i v a t i v e s have been shown to d e c r e a s e the number o f c o lifo rm organism s end i n c r e a s e th e number o f e n t e r o c o c c i in the i n t e s t i n a l f l o r a o f r a t s ( 5 l ) 8ad. human b e in g s (73)* I t seems l i k e l y t h a t a s i m i l a r s i t u a t i o n might a r i s e i n t h e rumen o f a c a l f supplem ented w ith s u lf a g u a n id in e . Since i t h a s been d e m o n stra te d t h a t c o b a l t i s c o n c e n tr a t e d in the m icro­ organisms p r e s e n t i n th e rumen o f th e sheep (127) i t i s p r o b a b le t h a t a s im i l a r c o n c e n t r a t i o n o c c u r s in t h e rumen o f th e c a lf* A s h i f t i n the p r e ­ dominant ty p e s o f organism s p r e s e n t i n th e rumen u n d e r th e s u lf a g u a n id in e i s p r o b a b l e . in flu en c eof I t i s e n tir e ly p o s s ib le th a t a s h if t of t h i s s o r t c o u ld i n f l u e n c e th e m i c r o b i a l c o b a l t m etabolism . V a r i a t i o n s in the r e s i s t a n c e t o d i g e s t i o n e x h i b i t e d by v a r i o u s ty p e s o f rumen m icroorganism s (103) c o u ld a l s o account f o r t h e e f f e c t o f s u lf a g u a n i d i n e su p p le m e n ta tio n on th e s u p n o s i ti o n t h a t s u lf a g u a n i d i n e r e s i s t a n t m icroorganism s a r e p r e ­ d o m in ately r e s i s t a n t to d i g e s t i o n . I n t h i s way th e c o b a l t co uld be concen­ t r a t e d i n th e m icro rog an ism s from t h e rumen and in a l a r g e measure be e lim ­ i n a te d a s a component p a r t o f the m ic r o b ia l c e l l s in the f e c e s o f the animal t h u s l e s s e n i n g th e e f f e c t o f th e in g e s t e d c o b a l t on th e b lo o d p i c t u r e by re d u c in g i t s a v a i l a b i l i t y f o r a b s o r p t i o n . C obalt i n c o n c e n t r a t i o n s v a r y i n g from 1 to 100 p a r t s p e r m i l l i o n has been snown to i n h i b i t growth and r e p r o d u c t i o n in L a c t o b a c i l l i ( 123) and o th e r r e p r e s e n t a t i v e s p e c i e s o f b a c t e r i a ( l l 2 ) . In v iv o ex p erim en ts (87) have shown t h a t o r o t e i n s y n t h e s i s by rumen f l o r a i s red u ced i n the p r e s e n c e o f 10 p a r t s p e r m i l l i o n c o b a l t a lth o u g h some p r o t e i n s y n t h e s i s o c c u rre d a t - 51 te n tim e s t h i s c o n c e n tr a tio n o f c o b a lt . I n h ib i t i o n o f b a c t e r i a by c o b a lt in tim a te s t h a t th e s e org an ism s ab so rb th e c o b a lt s in c e an i n t e r r u p t i o n must o cc u r in some p h ase o f t h e i r m etabolism f o r i n h i b i t i o n to o c c u r. T h is in t e r r u p t i o n o f th e b a c t e r i a l m etabolism co u ld be due to th e i n h i b i t o r y e f f e c t o f c o b a lt on o x id a tiv e enzymes in th e b a c t e r i a l c e lls * Levy et_ a l . (7 7 ) have d e m o n strated t h a t c o b a lt i n h i b i t s th e a c tio n o f s e v e ra l o f the o x id a tiv e enzymes b u t n o t th e p h o s p h o ry ly tic enzymes s tu d ie d in r a t l i v e r end k id n e y hom ogenates a t a c o n c e n tra tio n , o f 59 p a r t s p e r m illio n , the o n ly c o n c e n tr a tio n u s e d in th e in v e s tig a tio n * may have th e same e f f e c t . Lower c o n c e n tr a tio n s o f c o b a lt In c o n tr a s t to th e e f f e c t o f c o b a lt on o x id a tiv e enzymes i t h a s been shown to double th e a c t i v i t y o f arg in e.se a t a concen­ t r a t i o n o f 3b- p a r t s p e r m illio n (6 3 ) and to a c t i v a t e s tr o n g ly g ly c y lg ly c in e d ip e p tid a s e a t a c o n c e n tr a tio n o f 59 p a r t s p e r m illio n ( l l 6 ) . I n h ib it io n o f th e b a c t e r i a c o u ld o c c u r e i t h e r a s th e r e s u l t o f i n h i b i t i o n o f one or more o f t h e i r enzymeB o r by an a c c e le r a tio n o f th e a c tio n o f one o r more enzymes, i n e i t h e r case by thro w ing th e m etabolism o f th e b a c t e r i a l c e l l o u t o f b a la n c e , The p o ly c y th e m ia w hich w as produced i n the young c a lv e s was n o t main ta in e d a t i t s o r i g i n a l l e v e l a f t e r th e c a lv e s re a c h e d th e approxim ate age o f 30 weeks even though su p p le m e n ta tio n was c o n tin u e d a t i t s o r ig i n a l r a t e on a w eig h t b a s i s . The s e v e r i t y o f th e p o ly cy th em ia was l e s s in th e p r e ­ lim in a ry group composed o f an im als more advanced in a g e . A somewhat l e s s e r r a t e o f c o b a lt su p p le m e n ta tio n f a i l e d to produce a p o ly cy th em ic c o n d itio n in y e a r l i n g h e i f e r s . T h is may be due to th e f a i l u r e o f th e young an im als to d ev elo p a norm al rumen m ic r o f lo r a u n d er th e in f lu e n c e of th e heavy c o b a lt - 52 su p p le m e n ta tio n . I t h a s "been shown t h a t some c a lv e s f a i l to d ev elo p a normal rumen m ic r o f lo r a u n t i l th e y a r e o v e r 90 d ay s o f age (10^ ) • As h a s been in d ic a te d above, th e rumen m icro o rg an ism s msy be f u r t h e r su p p re sse d by th e h ig h c o n c e n tr a tio n o f c o b a lt i n th e rumen even a f t e r t h i s a g e . The p o s s i b i l i t y i s s tr o n g t h a t t h e r e i s a c o n c u rre n t in c r e a s e in th e rumen m ic ro f lo r a w ith th e lo w e rin g o f th e r e d b lo o d c e l l c o u n ts e n c o u n te re d a s th e a n im als become o l d e r . The r e s u l t s o b ta in e d w ith one an im al, C711, seem to su p p o rt th e v e l i d i t y o f t h i s h y p o th e s is s in c e th e w ith d ra w al o f s u l f e g u a n id in e from th e r a t i o n c o in c id e d w ith a d e p r e s s io n o f th e p o ly c y th e m ia . I t i s g ra n te d t h a t t h i s d id n o t o ccu r in t h e c a s e s o f th e o th e r two an im als in Group 3 b u t n e i t h e r was any m easurem ent o f rumen m ic r o f lo r a made on th e s e an im a ls- A gain, s in g le c a s e s a re i n s u f f i c i e n t ev id en ce f o r d raw in g any con- e lu s io n . Schade (112) h a s shown t h a t th e e f f e c t i v e n e s s o f a g iv e n c o b a lt c o n c e n tr a tio n f o r i n h i b i t i o n o f grow th i n P r o te u s v u l g a r i s c u l t u r e s i s a f u n c tio n o f th e number o f c e l l s p r e s e n t . T h is p ro b a b ly e x p la in s J o s la n d ’ s i n a b i l i t y t o p ro d u ce a p o ly cy th e m ia in 3 o f th e 4 ev es d ren ch ed d a i l y w ith c o b a lt f o r seven m onthsThe i n a b i l i t y o f th e c o b a lt supplem ented c a lv e s to grow et a r a t e eq u a l to t h a t i n R a g s d a le ’ s s ta n d a rd in c o n tr a s t to th e f a s t e r g a in diown by th e c o n tr o l group s e rv e s to in d ic a te t h a t th e l e v e l o f c o b a lt su p p le ­ m e n ta tio n en p lo y ed i n t h i s stu d y was t o x i c . The y e a r l i n g h e i f e r s (Group 6) , supplem ented a t a somewhat l e s s e r r a t e , g a in e d w eig h t a t a r a t e eq u al to t h a t o f t h e i r c o n t r o ls (Gro^p 5 )• T h is i s i n agreem ent w ith th e c o n c lu s io n o f K eener &t al.. (70) who found t h a t grow ing d a i r y an im als can t o l e r a t e up to 0 .5 m illig ra m c o b a lt d a i l y p e r pound body w eig h t over lo n g p e r io d s o f tim e . - 53 - D airy c a lv e s can t o l e r a t e s u lf a g u a n id in e a d m in is tr a tio n a t th e r a t e o f 2 p e r c e n t o f t h e i r d r y m a tte r in ta k e a s shown in t h i s stu d y f o r p e r io d s o f 39 and 125 d ay s, r e s p e c t i v e l y . o f th e recommended t h e r a p e u t i c d o sa g e . T h is i s c o n s id e ra b ly in e x c e s s Johnson e t al_. ( 6^) have shown th a t th e a d d itio n o f 1 p e r c e n t s u l f a t h a l i d i n e to th e r a t i o n o f 9 weeks o ld c a lv e s had no e f f e c t on grow th o r food consum ption a lth o u g h th e b a c t e r i a l count in th e f e c e s was d e c re a s e d a b o u t 80 p e r c e n t. There was no evidence o f any in c re a s e d s u s c e p t i b i l i t y to i n f e c t io n su b se q u e n t t o th e w ith d ra w al o f th e s u lfa g u a n id in e supplem ent- SUMMARY AND CONCLUSION • S eventeen d a ir y c a lv e s betw een th e eges o f 12 and 191 days were a l l o t t e d to 5 g ro u p s . One group r e c e iv e d no supplem ent and th e rem ain­ in g g ro u p s r e c e iv e d d a i l y supplem ents o f c o b a lt a t th e r a t e o f 1 m i l l i ­ gram p e r pound body w e ig h t, s u lfa g u a n id in e a t th e r a t e o f 2 p e r c e n t o f th e d ry m a tte r in ta k e o r a co m b in atio n o f th e p re c e d in g two supplem ents. The g ro u p s r e c e i v i n g s u lf a g u a n id in e w ere c o n tin u e d on th e c o b a lt supple­ ment a f t e r s u lf a g u a n id in e su p p le m e n ta tio n was d is c o n tin u e d . E ig h t y e a r­ l i n g h e if e rB were a l l o t t e d t o two g ro u p s, one r e c e iv in g a d a i l y c o b a lt supplem ent a t th e r a t e o f 0 .5 m illig ra m s p e r oound body w e ig h t and th e o th e r r e c e i v in g no su p p lem en t. The d a i l y c o b a lt supplem ent was r e v is e d upward to 0 .7 5 m illig ra m s p e r pound a f t e r 8h days o f su p p le m e n ta tio n and c o n tin u e d f o r an a d d i t i o n a l 28 d ays. The re d b lo o d c e l l c o u n ts o f c a lv e s r e c e iv in g o n ly th e c o b a lt su p p le­ ment s t a r t i n g i n t h e i r second month o f l i f e le v e le d o f f a t ap p ro x im a te ly 20 m il l i o n s p e r c u b ic m illim e te r i n 3 o f th e k cases* C oncurrent in ­ c r e a s e s in hem oglobin c o n c e n tr a tio n s and re d b lo o d c e l l volum es o c c u rre d b u t a t a l e s s e r r a t e o f in c r e a s e , f i n a l l y re a c h in g e q u iv a le n t l e v e l s ra n g in g from 15 to 20 grams p e r c e n t and from k6 to 57 p e r c e n t, r e s p e c t iv e ly No change a t t r i b u t a b l e to su p p le m e n ta tio n was n o te d i n l e v e l s o f t o t a l serum p r o t e i n s , serum album in, p lasm a calc iu m .p la sm a in o rg a n ic p h o sp h o ru s, plasm a magnesium, and p lasm a a s c o r b ic a c id . S u lfa g u a n id in e su p p le m e n ta tio n p ro d u ced no a p p a re n t change in th e - 55 b lo o d p i c t u r e . When c o b a lt su p p le m e n ta tio n fo llo w e d o r was in c o n ju n c tio n w ith s u lf a g u a n id in e a d m in is tr a tio n , c o n c u rre n t and e q u iv a le n t in c r e a s e s o c c u rre d in r e d b lo o d c e l l c o u n ts , hem oglobin c o n c e n tr a tio n s , and re d b lo o d c e l l volum es b u t n o t t o a s g r e a t an e x te n t a s e n c o u n te re d w ith an im als supplem ented w ith c o b a lt o n ly . The s e v e r i t y o f th e p o ly c y th e m ia d e c re a s e d a s th e a n im a ls approached th e age o f 30 weeks and a n im a ls w hich were ap p ro a c h in g t h i s age a t the tim e when c o b a lt su p o le ro e n ta tio n was s t a r t e d r e a c te d in a l i k e manner. No r e ­ sponse t o c o b a lt su p p le m e n ta tio n was n o te d in th e y e a r lin g h e i f e r group. The p o s s i b i l i t y was d is c u s s e d t h a t a s th e rumen m ic r o f lo r a becomes w e ll e s ta b li s h e d and assum es norm al n u m e ric a l p r o p o r tio n s th e p o ly cy th em ic e f f e c t o f c o b a lt becomes l e s s se v e re and f i n a l l y re a c h e s a p o i n t o f no consequence. Subnormal w eig h t g a in s were e n c o u n te re d in a l l c a lv e s r e c e iv in g th e c o b a lt su p p lem en t. d a l e ’ s s ta n d a rd . The c o n tr o l c a lv e s g a in ed a t a r a t e in e x cess o f Rags­ No d if f e r e n c e i n r a t e o f g a in betw een c o b a lt supplem ented and c o n tr o l y e a r l i n g h e i f e r s was n o te d . I t i s co n clu d ed t h r t c o b a lt i s a b le to induce a p o ly cy th em ia in c a lv e s a t a tim e when t h e i r rumen m ic r o f lo r a i s n o t y e t w e ll e s ta b lis h e d . i - 56 - LITERATURE CITED (1) A nderson, H. D ., Underwood, E. J . , end E lvehjem , C. 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C ., an d O rten , J . M. 1948 C holine and th e P ro d u c tio n o f P o ly cy th em ia by C obalt in th e R a t. Amer. J o u r . P h y s io l. 154: 513-516* - 58 (23) B u c c ie ro , M. C ., and. O rten , J . M. 1949 The E f f e c t o f C o b alt on th e Oxygen C ap a city and th e Methemog lo b in C ontent o f th e B lood. Blood 4 : 395-397* (2*0 B urk, D ., Schade, A. L . f H e sselb ack , H. L ., and F is c h e r , C. E. 1946 C o b alt I n h ib i t i o n o f T issu e R e s p ir a tio n , G ly c o ly s is , and Growth. F ed. P ro c . 5 J 126-127- (25) Chang, Y. T -, Chen, J . M., and Shen, T. 1943 Induced P o ly cy th em ia i n Salam ander by C o b a lt, A sc o rb ic Acid and O th er W ater-S o lu b le V ita m in s. A rch. Biochera. 3 : 253-259- (26) C lark ,. E . P . , and C o llip , J . B. 1925 A Study o f th e T i s d a l l Method f o r th e D e te rm in a tio n o f Blood Serum Calcium w ith a S uggested M o d if ic a tio n . J o u r . B io l. Chem. 6 3 t 46 1 -4 6 4 . 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M. 1938 The In flu e n c e o f C obalt on P ine D isease in Sheep. J o u r . 3 6 : 1800-1805. Ai Biochem. (32) C o rn il, L ., P o u r s in e s , Y ., and D um ont-Lagre, Mme1943 H e p a t i t i s in G uinea P ig s P o iso n ed w ith C obalt A c e ta te . In ­ flu e n c e o f Splenectom y on Development o f th e L e s io n s . Soc. de B iol* ( P a r i s ) Compt. Rend. 137* 459-460- Abs. Chem. Abs3 9 : 2569 (1 9 4 5 ). (33) D a ft, F» S ., A shburn, L . L ., and S e b r e ll, W. H. 1942 B io tin D e fic ie n c y and O th er Changes in R a ts Given S u lfa n ily lg u a n id in e or S u ccin y l S u lf a th ia z o le in P u r if i e d D ie ts S cience 9 6 : 321-322- ^ - 59 (3*0 (35) (36) D e ft, P . S ., and S e b r e ll, W# H. 1945 S ulfonam ides and V itam in D e fic ie n c ie s * 3 : 1+9-72. V itam in s and Hormones Dann, W. J . 191+1 The S y n th e s is o f N ic o tin ic Acid by th e R a t. l 4 l ! 803- 808. D av is, J . E . 1937 C o b alt P o ly c y th e m ia in th e Dog. 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The E f f e c t o f M etals on the A ctivity o f th e Rumen B a c te r ia . Biochem. J o u r . *4-6: 36-*43« (88) McNaught, E. J . , and P a u l, U.W. 1939 S easonal V a r ia tio n in C obalt C ontent o f N orth I s la n d P as­ t u r e s . New Z e a l. J o u r . S c i. and T ech n o l. 21 (B ): 9 5 -1 0 l.A b s. Chem. A bs. 3*4-: 2511. (19*40). (89) M ic h a la z z i, A. M., and S aviano, M. 19*40 I n v e s t i g a t i o n s on th e P o ly cy th em ia from C o b a lt. A rch. S c i. B io l. ( I t a l y ) 26: 3*41-361. Abs. Chem. A bs. 35! 8108. (l9*4l) (90) M in d lin , R . L ., and B u tle r , A. M. 1938 The D e te rm in a tio n o f A sco rb ic Acid in Plasm a; a Macromethod and a M icrom ethod. J o u r . B io l. Chem. 122: 673-686. - 63 (79) L ig h t, E . P ., C rec ae , L . J . , O lc o tt, C. T ., and P rey , C. N. 1942 I n h i b i t i o n o f th e S ym biotic S y n th e s is o f B Complex P a c to re by S u lfo n am id es. J o u r . N u tr. 24: 427-435* (80) M ackenzie, J . B ., M ackenzie, C. G ., end McCollum, E. V, 1941 The E f f e c t o f S u lfa n ily lg u a n id in e on th e T hyroid o f th e E a tS cience 9 4 : 518-519• ( 81 ) M a rs h a ll, E. K ., J r . , B r a tto n , A. C ., W hite, H. J . , and L i t c h f i e l d , J.T. 0 . 1*1*1* 0 . 1*61 0 .3 3 1 0 .2 8 1 0.1*58 0.1*22 0.1*03 0 .3 8 2 o.i*66 0-5 8 7 0.1*22 0 .5 1 2 0 .3 8 1 0 .3 8 9 0 .3 8 0 0.1*76 0 .2 0 8 0-3 5 9 0 .3 3 6 0.1*96 0 . 1*06 0.1*82 0 .5 3 7 0 .5 5 6 0.1*85 0 . 5*1* 0 . 31*1* Appendix T able 2 - C ontinued Blond P ic t u r e o f Animal Number C68l—P r e lim in a r y Group Whole Blood___________________ Bed Blood HemoBed Blood Mean C orpusC e ll Count g lo b in C e ll V ol. c u la r Volume Ace days 36 b 371 378 385 392 399 b06 1*13 1*20 1*2? 1*33 bbo 1*1*7 l*5b 1*67 b75 1*82 1*89 1*96 M/mm^ --------- 10.90 12 .2 0 10 .88 10.20 10.18 8 .6 0 7 .9 7 7 .0 5 7 .3 0 6 .2 2 9 .1 0 8.C8 7 .9 5 6 .6 9 7 .6 6 6 .81 6 . ll* gm $ 1 5 *1*0 15*50 13*60 13*35 1 1 .8 5 11.60 11.23 1 0 .2 7 10*27 9*55 9 .5 5 9 .0 3 11 .bo 9.83 9 .6 3 9.13 1 0 .2 7 1 0 .3 5 9*83 56 uJ Mean C orpusc u la r Hemoa lo b in uug 3 6 .5 b o .o ---------- —---- -- 3 b .5 3b.0 2 9 .0 29.0 28.0 25*5 2 7 .0 2 3 .5 2 b .0 2 2 .5 27.0 26.0 26.0 2 6 .5 2 8 .5 25*5 2b . 5 31*7 27*9 26.7 2 8 .b 27*5 29*7 33*9 33*3 3 2 .9 3 6 .2 2 9 .7 3 2 .2 3 2 .7 3 9 .6 3 7 .2 37*b 3 9 .9 1 2 .5 1 0 .9 1 0 .9 1 1 .b 1 1 .0 1 1.9 1 2 .9 13-5 13*1 lb .5 1 2 .5 1 2 .2 1 2 .1 13*6 13.b 15*2 1 6 .0 Mean -C o rp u sc u la r Hemog lo b in Cone • i b 2 .2 3 8 .8 3 9 -b 39*3 bo .9 b o .o b o .i b o .3 3 8 .0 b o .6 39*8 b o .l b-2.2 3 7 .8 37*0 3b. 5 36.0 b o .6 b o .l P la s m a ________________ C al- I n o rg . P h o s- Kagne- A sco rb ic cium p h o ru s sium A cid mg i> 1 0 .7 1 1 .5 1 1 .6 1 2 .0 11.3 1 1 .6 11.3 1 1 .6 1 2 .0 1 2 .1 1 1 .6 11.b 1 0 .7 1 1 .6 1 0 .6 1 0 .7 1 1.3 1 1 .0 9 .5 mg $ mg £ 6 .19 7*58 6 .5 1 7*67 7.31 6.1 9 9*13 7-91 7*71 8 .0 6 7*81 6 .5 1 7 .6 2 6 .51 8 .0 6 6 .9 5 7.81 6 .3 5 7 .b b 2.11 2.30 2 .8 2 1*55 2 .2 5 1*57 1.73 2.63 2*15 2 .7 5 2.b 9 2 .0 2 3 .3 6 1 .7 6 1 .6 8 2 .0 0 2 .0 8 1 .8 8 1 .7 1 rng ; 0 .b l2 0.b 7 9 0.500 0 . 36b 0 .3 8 7 0.b 9 9 0.bb8 0 .2 9 7 0 .b o 5 o.b 53 0 .3 6 6 0.318 0.391 0.363 0 .3 6 5 0 .3 5 6 0 .5 6 6 0 .3 b b o .b o b Appendix T able 3 Blood P i c tu r e o f Animal Number C684— P r e lim in a r y Group Ace days 161 168 182 189 196 202 210 217 224 231 237 245 252 259 266 273 280 287 292 301 308 315 322 329 Whole Blood____________________ Bed Blood HemoBed Blood C e ll Count glob in C e ll V ol. Mean Corpus­ c u l a r Volume gm i u^ M/mm-5 11.20 1 0 .5 5 14.30 1 3 .79 1 4 .4 9 1 1 .3 5 —-----— —-----— 1 2 .7 2 1 6 .6 3 ---------- 13-30 ---------- 15.80 15*12 13-95 1 3 .7 3 14.80 15-10 1 4 .0 5 — 1 4 .5 0 13-17 13 .0 0 U .60 12.60 15-20 14.80 14-55 1 4 .1 0 1 3.60 1 5.60 1 5 .00 16.50 1 6 .20 17.10 17.50 17.10 l6 .i« ) 16.80 1 6 .2 5 16.80 1 7 .20 16.80 17.20 17.50 % 3 4 .7 33*5 3 0 .5 31-0 4 2 .0 4 1 .5 3 7 .0 3 8.0 3 6 .0 3 9 .5 4 1 .0 4 2 .0 4 3 .0 4 8 .0 4 3 .5 4 6 .0 4 5 .0 4 3 .0 4 2 .0 4 3 .0 4 5 .0 4 3 .0 4 1 .5 4 8 .0 A Mean Corpus­ c u la r Hemo­ g lo b in uug _____ —---- — --------— 2 7 .2 2 9 .4 2 7 .6 30-1 2 5 .5 3 3-5 1 0 .4 1 1 .9 1 0 .6 1 0 .7 1 0.0 1 2 .4 —— — ——----- —------- —---- — 3 2 .2 25.3 1 1 .8 9 .9 ---------- ------- -- 3 6.1 1 2 .9 —-----— — — 29-1 29.8 3 0 .8 3 0 .6 29.1 2 9 .8 3 0 .6 1 0 .8 1 0 .8 12 .0 11-8 1 1 .4 1 1 .4 12.0 ------- — — — 3 3.1 1 2 .1 Mean Corpus­ c u la r Hemo­ g lo b in Cone. * Cal­ cium Plasma In o rg . Phos­ p h o ru s mg f> 3 8 .0 3 8 .8 3 8 .0 4 0 .6 3 6 .2 3 5 .7 39-3 3 7 .1 3 7 .8 3 9 .5 3 6 .6 39.3 37.7 3 5 .6 4 0 .2 3 7 .2 3 6 .4 3 9 .1 3 8 .7 39-1 3 8 .2 39 .1 4 1 .4 3 6 .5 1 1 .2 1 1 .0 1 1 .1 1 1.0 1 0 .9 8 .8 1 0 .6 1 1.3 1 1 .3 11 .1 9 .5 1 0 .0 1 0 .5 1 0 .7 1 2 .3 1 1 .7 1 1 .6 1 0.8 1 0 .6 1 1 .0 1 1 .3 1 0 .6 1 0 .6 1 2 .6 Magne­ sium A scorbi Acid mg i mg mg $ 7 .9 6 8.93 6 .19 6 .3 5 7 .8 6 5*87 6 .4 8 6 .1 9 7 .9 1 7 .0 2 5 .48 6.83 8.33 7-10 7 .5 8 7.27 5-79 6.79 7-81 5*81 6 .3 2 5 .81 8 .8 1 7.96 2.09 2.23 1 .7 0 2.03 1 .8 8 2.40 1 .8 7 2 .1 5 2 .0 6 2 .0 4 1.53 1 .9 8 2 .2 5 1-59 2.00 2.23 1 .9 2 1 .8 1 1 .9 8 2 .7 5 2 .06 2 .02 2 .04 1 .1 4 0.353 0 .4 2 8 0 .1 8 1 0.361 0 .1 9 4 0 .4 8 5 0 .332 0.2 8 1 0.0 8 8 0.280 0.3 7 4 0.3 1 3 0 .3 9 3 0.413 0.4 7 6 0.447 0.373 0-496 0.5 8 0 0.551 0.5 2 2 0.3 2 1 0 .5 6 4 0.4 1 2 Appendix T able 3 - C ontinued Whole Blood Red Blood Hemo­ C ell Count g lo b in Age days 336 3^3 350 357 364 385 392 399 413 419 426 433 440 Bed Blood C ell V o l. M/mm^ gm i 'P 12.80 17.50 17.70 18.50 17.60 18.80 17.90 1 8.10 1 7.50 1 7 .0 0 1 6 .50 1 5 .1 0 15*60 1 3 .3 5 1 3 .77 4 5 .5 4 8 .0 4 5 .0 4 6 .5 4 3 .0 4 5 .5 4 2 .5 4 l.O 4 2 .0 3 7 .5 4 0 .0 3 6 .0 3 4 .5 --------- 16.10 18.10 13-55 1 4 .1 6 1 3 .5 5 12.80 1 4 .0 2 1 1 .9 5 ----- -- Mean Corpus­ cu lar Volume U -' Mean Corpus­ cu la r Hemo­ g lo b in uug 3 5 .5 27.4 ------ 1 3 .8 1 0 .6 —---- —— — -------— 2 6.7 25.1 31.4 29.0 3 1 .0 2 0 .3 25-5 3 0 .1 1 1.1 1 0 .0 1 2 .9 1 2.0 1 2 .2 1 1.8 11.1 1 1 .2 ------ —— Mean Corpus­ cu lar Hemo­ g lo b in Cone. $ 3 8 .9 3 8 .5 3 9 .1 3 8 .8 4 1 .6 3 9 .8 4 1 .2 4 1 .5 3 9.3 40-3 39.0 3 7 .1 3 9 .9 Cal­ cium Plasma Inorg. Phos­ phorus %£ I BCD 1 Blood P i c tu r e o f Animal Humber C634— P r e lim in a r y Group A scorbic Acid mg i mg % mg f, mg % 1 0 .9 9 .5 9 .7 9 .3 1 1 .5 9 .6 1 0 .7 1 1 .6 1 0 .5 12.0 1 0 .5 1 2 .2 1 1 .2 6 .7 2 6 .6 9 6 .4 8 7 .6 2 7-35 5.63 7 .6 2 7 .7 1 7 .4 4 6 .4 4 6 .2 5 7 .6 7 6 .2 2 1 .4 8 2.1JO 2 .1 6 2.11 2 .2 5 2.03 2 .2 4 2.10 2.99 2 .7 5 2 .63 3 .9 6 1.70 0.390 0 .3 8 4 0 .3 5 5 O .38I 0.2S9 0-584 0 .511 0 .4 8 4 ——— 0.3 4 2 0.270 0.211 0.193 Appendix Table 4 Blood P i c tu r e o f Animal Humber C718—Group 1 45 53 6o 67 74 81 88 95 102 109 116 123 130 137 144 151 158 165 172 179 186 193 200 207 214 221 9-35 7 .4 5 8 .5 5 7-78 9-48 1 0 .4 4 9 .2 4 IO .58 8 .4 0 9 .0 3 7 .9 5 7 .5 3 9 .3 5 7 .7 1 9 .1 5 9-10 8 .2 2 8 .2 2 8 .7 5 7-87 9 -39 8 .8 5 8 .3 0 7 .6 4 6 .8 2 8 .1 4 9 .7 7 8.90 9 .2 3 9 .4 0 10.80 11.60 1 0 .13 10.63 10.20 9.63 1 0 .0 7 9.33 1 0 .4 3 1 1 .93 1 1 .0 7 1 0 .8 7 1 0 .1 3 1 0 .3 5 11.00 10.43 1 2 .4 5 1 2 .4 5 1 0 .93 10.13 1 0 .93 1 0 .5 7 2 8 .5 28.0 24.0 2 4 .0 2 6 .5 28.0 25.0 29.0 2 7 .5 2 5 .5 2 7 .5 2 3 .5 29.0 3 2 .0 2 7 .5 28.0 26.0 2 6 .5 26-5 2 5 .0 3 3 .0 2 9 .5 26.0 25.0 2 7 .5 2 5 .0 3 0 .5 37-6 28.1 3 0 .8 28.0 26.8 27.1 2 7 .4 3 2 .7 2 8 .2 3 4 .6 3 1 .2 31-0 4 1 .5 3 0 .1 30-8 3 1 .6 3 2 .2 3 0 .3 3 1.8 35-1 3 3 .3 31-3 3 2 .7 4 0.3 3 0 .7 1 0 .4 1 1 .9 1 0 .8 1 2 .1 1 1 .4 ' 1 1 .1 1 1 .0 1 0 .0 1 2 .1 1 0 .7 1 2 .7 1 2 .4 1 1 .2 1 5 -5 1 2 .1 11-9 12.3 1 2 .6 1 2 .6 13-3 13-3 1 4 .1 1 3 .2 13.3 1 6 .0 13.0 3 4 .3 3 1 .8 3 8 .5 3 9 .2 4 0 .8 4 l.4 4 0 -5 3 6 .7 37-1 3 7 .8 3 6 .6 3 9 .7 36.0 3 7 .3 40-3 3 8 .8 3 9 .0 3 9 .1 4 1 .5 4 1 .7 3 7 .7 4 2 .2 4 2 .0 4 0 .5 3 9 .7 4 2 .3 Serpm Plasma T otal Albu- Cai_ Inorg. MagA scorbic Pro- min cium Phos- nesiura Acid p h o ru s te in gm £ e® -p mg i mg $ mg % mg i ——— ------- — -------- — ---- — — — -------- — -----— — 506 5-51 5 .0 6 5-19 5-72 5 .0 2 6 .21 5 .83 5 -6 5 — . - 3 .7 4 3-41 3*36 3 .1 6 3 .0 8 3-63 3-48 3-61 ---------- ---------- ---------- — ---- 5-49 5-53 3*51 3-53 ------- ---------- 6 .3 7 3*92 --- ----- ---------- ---------- . . . . ---------- 6 .4 8 5-88 3*66 3*64 1 0 .1 1 0 .8 10.4 1 1 .3 1 1 .9 1 1 .7 1 1 .1 1 1 .0 ---------- 1 1 .2 11 -5 1 0 .? 1 1 .5 1 2 .7 1 1 .3 1 1 .8 11.1 12.0 11 .9 1 1 .4 1 1 .5 12.0 1 1 .2 11.1 11.3 1 1 .9 6 .69 7 .2 7 5.61 6 .5 4 8 .8 1 8 .1 7 8 .8 1 1 0 .1 6 6 .3 3 6.58 7 .1 0 7.81 6 .4 1 6 .8 3 6 .9 5 7 .67 1 0.16 7o3 7 .4 4 7 .9 6 9-19 7 .3 5 7-58 7 .3 6 6 .3 5 7.27 2.20 2 .15 1 .2 0 1.91 2.37 1 .6 8 2.00 1 .5 8 3-83 2.3 2 2 .0 4 2.23 2.66 3 .9 5 2.17 3.00 2.2 5 2.6 2 2.31 I .83 1 .9 4 3-20 2.58 1 .8 1 2.38 2.06 0 .4 8 4 0.549 0.5 4 1 0.571 0.680 0.570 0 .6 ? 4 0.6 1 0 0.5 6 1 0.4 2 7 0 .4 9 2 0.613 0.3 4 6 0 .4 5 3 0 .5 7 2 0 .5 9 5 0.5 6 1 0 .5 6 2 0 .6 0 2 C-303 0.6 4 1 0.580 0 .4 0 2 0.6 0 8 0 .5 4 5 0.553 “9 L - Whole Blood________________ Red Blood Hemo- Red Blood Mean Corpus- Mean Corpus- Mean CorpusC ell Count g lo - C ell V ol. cular Volume cu la r Hemo- cu la r Hemog lo b in g lo b in Cone, b in Ase M/mm3 u^ gm $ uug days 2 Appendix T sb le 5 Blood P i c tu r e o f Animal Humber C719— Group 1 Age Whole Blood Red Blood Hemo­ C e ll Count g lo ­ b in d ays M/mm3 gm £ 18 25 38 1+6 53 6o 67 71+ 81 88 95 102 109 116 123 130 137 ik k 151 158 165 172 179 186 193 10.50 10-30 10.20 8 .8 0 8 .7 0 8 .7 0 9 .3 6 8 .7 7 8 .7 5 8 .0 9 8.33 8 .7 6 6 .8 0 7.50 9 .2 0 8 .0 8 8 .1 0 7*64 8 .3 9 7-85 9-1+0 8 .7 5 8 .1 2 6.50 7 .6 5 9*90 1 2 .90 1 2.53 1 0 .57 1 0 .93 1 1 .8 5 1 2 .6 5 1 1 .50 11.50 1 1 .37 1 2 .7 7 11.23 10.13 11.30 13-1+3 11 .6 7 11.50 11 .23 12.00 12.10 12.90 1 1 .8 5 12.1+5 12.53 12.10 Red Blood C e ll Volume Mean Corpus­ c u la r Volume i u5 2 k. 5 3 6 .0 3 6 .0 33-0 31.0 2 9 .0 3 3 .0 3 2 .0 26.0 2 6 .5 3 2 .0 3 1 .0 2 5 .0 2 9 .5 3 5 .5 3 1-5 3 2 .5 28.0 3 0 .0 3 0 .0 3 3 .5 3 0 .0 32.0 3 0 .5 31-5 23.3 3 5 .0 350 3 7 .5 35*6 33 0 3 3 -5 3 6 .5 2 9 .7 3 2 .8 33 , k 3 5 -k 3 6 .8 390 3 8 .6 39-0 k o .i 3 6 .6 35-8 3 8 .2 3 5-6 3^0 39. k 1+6 .9 1+1.2 Mean Corpus­ c u la r Hemoglo­ b in uug 9.1+ 1 2 .5 1 2 .3 1 2 .0 1 2 .6 1 3 .6 1 2 .8 13*1 13-1 ll+ .l 150 1 2.8 11+.9 15-1 11+.6 14.1+ 11+.2 11+.7 11+.3 15*^ 1 3 .7 13-5 150 190 15. s Mean Corpus­ c u la r Hemoglo­ b in Cone. * 1+0 . 1+ 35-8 31+.3 3 2.0 350 1+0 .9 38.3 35-9 1+1+.2 1+2.9 3 9 .9 3 6 .2 1+0 :5 380 37-8 3 7 .0 3 5 . 1+ 1+0 .1 1+0 .0 1+0 .3 3 8 .5 3 9 .5 3 8 .9 1+1.1 3 8 .1+ Serum T o ta l Albumin P r o te in gm $ gm -------------------- - —--------- --------- --- --- - — — ----------- — ------------ —--------- ------------ ---------— 5.80 509 6.13 5 .7 2 5.53 5-63 6 .2 3 5.61 5-78 306 3 .2 5 3 .2 1 3.21+ 3-13 3 .2 6 3 .6 2 3 .1 5 3*1+3 —— — 5-98 6 .1 k 3 . 1+0 3 .5 5 ------- ------- 6.33 3*1+1 ------------ ------------ Calcium Plagm a In o rg . Magne slum Phos­ p h o ru s A scorbi A cid mg $> mg % mg $ mg $ llO 1 1.3 1 1 .2 1 2 .0 10 .8 1 1 .6 1 1 .2 11.1+ 1 1 .5 1 1 .9 1 1-5 1 2 .2 1 0 .5 1 1 .8 1 1 .5 1 2 .5 1 0 .9 11-7 1 1 .5 1 1 .8 1 0 .9 11-3 1 0 .5 1 2 .2 1 2 .7 7.53 6 .3 2 6 .01 6 .3 8 6 .3 2 5 .76 6 .1+8 7 .7 1 8 .50 3 .2 3 7 .9 1 7 .2 7 7.1+9 8 .1 2 7 .27 6 .6 5 7 .0 2 8 . 71+ 9 .5 5 7 .0 6 9 .0 6 7-91 6 .1+8 705 7-71 2 .2 5 1.63 1 .9 8 2.09 1 .9 7 1 .6 0 1 .9 3 1 .1 3 1 .9 0 1 .7 9 1 .9 8 202 2.08 1*50 2.33 3 .2 7 2 .06 2.1+0 1.91+ 2.02 1 .90 1*57 2.1 5 2 .5 7 3-22 0 050 0.1+59 0 . 1+60 0.1+19 0.1+66 0.1+21 0.5 1 9 O .5I+6 0.551+ 0.570 — ---------— 0 .5 8 6 0 .6 2 2 0.1+72 0.1+96 0.279 0.1+69 0.1+95 0.1+58 0.3 9 7 0.1+93 0 .2 9 5 0.1+75 0.1+99 0-336 .Appendix T able 6 Blood P ic tu r e o f Animal Number C724— Group 1 Age Whole Blood Bed Blood Hemo­ C e ll Count g lo ­ b in M/mrn-^ gm £ 32 39 46 53 6o 67 74 81 88 95 102 109 116 123 130 137 144 151 158 165 172 179 186 193 200 207 214 7 .8 6 7*40 8 .2 1 8 .4 5 7 .3 5 9-52 8 .4 4 8 .2 5 7 .6 5 10 .6 7 7 .8 5 10.30 7 .5 9 9 .0 7 8 .0 5 8 .3 5 10.50 10 .0 7 1 1 .1 5 1 1 .7 5 11 .67 1 2 .4 5 1 1 .3 7 1 1 .60 12.00 1305 11 .2 3 1 2 .8 3 12.00 11.30 1 1 .2 3 12.00 1 0 .8 ? 12.53 11.93 12.00 1 1 .8 5 1 2 .10 12.00 11.60 11.30 11.93 1 0 .5 7 — — 7 .7 4 9 .5 7 7 .4 5 8 .3 2 9 -12 8 .2 0 7 .5 0 7 .6 8 7 .5 2 7 .0 0 Mean Corpus­ c u la r Volume i u^ 3 0 .5 2 7 .5 3 0 .0 3 2 .0 3 3 .0 2 8 .5 30.0 3 1 .0 31.0 38.0 2 8 .0 3 4 .0 3 2 .0 2 8 .5 2 8 .5 2 9 .5 2 7 .5 3 1 .5 2 ?.0 29.0 3 2 .0 35-0 3 0 .5 3 0 .5 29.0 30.0 2 6 .5 Mean Corpus­ c u la r Hemoglo­ b in uug 3 8 .8 3 7 .2 3 6 .5 3 7 .9 4 4 .9 2 9.9 3 5 .5 3 7 .6 4 0 .5 3 5 .6 3 5 .7 3 3 .0 4 2 .2 3 1 .4 3 5 .4 350 13-4 1 3 .6 1 3 .6 1 3 .9 1 5 .9 1 3 .1 1 3-5 1 4 .1 15-7 1 2 .5 14.3 1 2 .5 15.s 1 2 .5 14.0 1 4 .4 --- --- ------- 4 0 .7 2 8 .2 3 8 .9 3 8 .5 3 8 .4 3 7 .2 4 o .7 3 7 .8 3 9 .9 3 7 .9 1 6 .2 1 2 .5 1 6 .1 1 4 .2 13.3 1 4.6 15-5 14.7 15-9 15-1 Mean Corpus­ c u la r Hemoglo­ b in Cone. £ 3 4 .4 3 6 .6 3 7 .2 3 6 .7 3 5 .4 43-7 3 7 .9 3 7 .4 3 8 .7 35-1 4 o .l 3 7 .7 3 7 .5 3 9 .6 3 9 .4 4 0 .7 39-5 3 9.8 4 4 .2 4 1 .4 3 7.0 3 4 .6 3 9 .3 3 8 .0 39.0 3 9 .8 3 9 .9 Serum T o ta l Albumin P r o te in Calcium Plasm a In o rg . Mag­ nesium Phos­ p h o ru s A sco rb ic Acid gm £ gm £ mg £ mg £ mg £ mg £ 5 .4 4 5.51 509 5.44 5-39 5.96 5 .56 6.00 3-38 3 .2 9 3-50 3-53 3-61 3-84 3 .60 ------------- ------------- 5.37 5.98 3 .4 3 3-74 --- —— ------- 5-81 5-58 3 .6 4 3-71 1 1 .7 1 1 .9 1 2 .0 1 0 .2 12 .0 1 1 .2 1 2 .1 11-9 1 2 .5 11-3 1 0 .7 1 0 .6 11 .0 1 1 .1 1 2 .5 1 3 .2 1 1-5 1 2 .3 1 0 .6 1 2 .0 11.3 1 2 .4 1 1 .2 11*3 11 -5 1 1 .4 11.3 8 .6 2 7*44 6.51 6 .2 5 7*86 5-63 7-10 6 .9 5 7 .7 1 8 .9 3 7.91 8 .3 9 7 .4 4 6.91 7.91 7.71 7 .5 8 6 .6 5 8 .6 2 7-71 9.33 1 0 .9 6 8 .8 7 7 .0 2 8 .5 0 9.40 8.33 1-53 1 .2 6 1 .7 6 1 .9 9 1 .9 4 2.07 2 .69 2 .0 2 2.30 2 .3 6 1 .2 6 2 .12 1-91 1 .8 8 1 .8 4 4 .0 8 1.8 3 2.06 1 .63 2.06 1 .9 4 3.10 2.30 2.36 2.24 1 .8 2 1 .2 8 0 .2 4 5 0.259 0 .2 7 2 0.3 5 9 0.440 0.306 0.279 0.4 2 8 0.3 9 3 0.2 8 7 O.365 0.453 0.371 0.4 1 9 O.368 0.3 2 3 0 .3 4 5 0.316 0.390 0.451 0 .3 6 7 0.213 0 .3 9 4 0.3 3 9 0.380 0.310 0.2 7 9 VPl 00• days Bed Blood C e ll Volume --- — ------- 6 .1 4 6 .0 7 5-98 6 .21 6 .2 1 6.19 6 .0 2 ------- 6 .2 6 6 .1 4 3*94 3 .9 4 3 .8 5 3-98 3-92 3 .3 3 4 .3 4 4 .1 2 4 .1 4 4 .0 8 .Appendix T able 7 Blond P ic tu r e o f Animal Number C742— Grotto 1 Whole Blood Bed Blood HemoC e ll Count s i n b in Age day b M/mnr' 33 7 .6 0 8 .6 4 9*45 9 .5 2 4o 47 54 6l 68 75 81 90 95 102 109 116 123 130 137 143 151 158 165 172 180 187 193 200 207 9 -4 2 10.50 10 .9 6 10.70 1 0 .6 5 9 .8 8 10.90 10.51 10 .1 4 1 0 .9 2 10 .57 1 1 .6 1 10.46 10-36 11.20 9 .9 9 -------------- 1 0 .4 5 10 .5 1 1 0 .5 5 9 .9 5 10.48 gm £ 9 .7 7 9 .7 7 10.13 9 .9 0 10.50 11.30 1 0 .5 7 10.93 11.23 1 2.10 1 2 .2 7 1 2 .60 11.60 1 2 .4 5 12.00 1 2 .4 5 11.50 10.50 1 2 .CO 1 1 .6 7 12 .8 3 1 1 .50 1 1 .8 5 11.60 11.50 1 1 .7 5 Red Elood C e ll Volume Kean Corpus­ c u la r Volume £ u^ 2 7 .0 26.0 28.0 2 7 .5 28.0 3 0 .5 2 9 .5 3 0 .5 31*5 3 3 -5 35-5 3 0 .1 2 9.6 2 8.9 29-7 29*0 2 6.9 2 8 .5 2 9 .6 33-9 2 9 .4 33*8 3 0 .6 2 9 .8 3 0 .3 28.9 3 0 .6 27.0 2 7 .2 30.0 3 2 .0 3 5 -5 3 1 .0 3 2 .5 3 2 .0 33-5 3 2 .0 28.0 3 0 .5 30 .c 3 2 .5 2 8 .5 29-0 3 0 .5 30.0 28.0 -------------- 27.3 2 7 .6 2 8.9 3 0 .2 26 .7 Kean Corpus­ c u la r Hemoglo­ b in uug 1 2 .9 11.3 1 0 .7 1 0 .4 1 1 .1 10-8 9 -6 1 0 .2 1 0 .5 1 2 .2 1 1.3 12.0 1 1 .4 1 1 .4 1 1 .4 1 0 .7 1 1 .0 1 0 .1 1 0 .7 1 1 .7 ——— 11.0 1 1 .3 1 1.0 1 1 .6 1 1 .2 Mean Corpus­ c u la r Hemoglo­ b in Cone£ 3 6 .2 3 7 .6 3 6 .2 3 6 .0 3 7 .5 37.0 35-8 35 -8 35-7 3 6 .1 3 8 .3 3 5 .5 3 7 .4 38.3 3 7 .5 3 7 .2 35-9 3 7 .5 3 9 .3 3 8 .9 3 9 .5 4 0 .4 4 0 .9 38.0 38.3 4 2 .0 Som n T o ta l Albumin P r o te in Calcium Pl?-.sma In o rg . Magnesium Phos­ p h o ru s A sco rb ic Acid gm £ gm £ mg $ mg £ mg £ mg £ 5-73 5 .6 4 5-78 6 .1 4 5*86 6 .1 3 5* 95 5-70 5-39 5-83 5.86 5-70 6 .13 5 .6 5 5*72 5*98 5.53 5.53 5.93 6 .2 8 6 .5 1 6 .6 1 6 .1 4 6 .6 3 6 .2 6 6 .30 3-51 3 .4 2 3 .5 6 3-88 3-79 3-3 6 3-6 4 3-55 3-29 3-51 3 .3 9 3 .5 0 —— 3 .4 2 3*42 3-70 3-40 3-29 3*53 3 .6 0 3 .20 3 .5 5 3 .3 6 3*83 2 .83 3-71 1 1 .9 1 1 .3 1 0 .7 1 0 .5 1 1.0 1 0 .4 11*5 11.0 11.3 1 0 .9 11.0 12.1 1 1 .8 1 1-5 1 0 .9 11-5 ll.l 1 0.7 1 0 .5 1 1 .2 1 0 .2 1 0 .9 1 0 .8 1 1 .6 1 0 .9 11.1 9 .2 6 7-44 9-77 7-91 7-58 8.23 11.1 6 7 .5 8 7 .9 1 9 .4 7 6.51 8 .3 ° 8 .0 6 8 .39 7 .9 6 7 .91 7.49 6 .89 6 .0 7 4 .3 5 5*14 7 .58 - 6.13 6.10 5.61 5-17 2.30 2.21 2.11 2.06 2 .4 2 2 .13 2.60 1 .7 6 2.31 1 .9 4 1 .9 8 2 .7 5 2 .1 ? 1 .8 8 1 .93 1 .80 2 .0 5 1 .8 8 1 .7 4 2 .0 4 2.51 1 .7 8 1 .8 5 2.88 1 .6 5 2.39 0 .4 4 5 0 .4 8 6 0 .6 7 3 0.640 0 .6 4 0 0 .5 4 6 0 .4 9 9 0 .7 0 0 0 .6 8 6 0 .6 8 4 0 .5 0 6 0 .6 4 7 0.613 0 .5 5 6 0 .4 9 4 0-582 0.498 0 .4 9 4 0 .5 0 3 0.296 0 .4 9 4 0 .3 3 9 0 .4 3 5 0 .5 9 5 0 .5 5 1 0-390 Appendix T able 8 Blood P ic tu r e o f Animal Number C749— Group 1 Whole Blood Red Blood Hemo­ C e ll Count g lo ­ b in Age days 75 82 89 96 102 111 116 123 130 137 144 151 158 1614172 179 186 193 201 208 214 221 228 M/mnP 1 0 .3 5 1 2 .0 7 13-33 9 .9 5 9 .7 5 8 .8 8 9.4-9 8 .8 8 9-13 8 .3 0 9 .5 7 9 .0 0 9 .3 5 8 .3 9 9 .1 2 10 .19 8 .3 4 10.00 9 .4 3 10 .CO 10 .0 8 7 .7 1 9 .6 2 Red Blood C e ll Volume Mean Corpus­ c u la r Volume gm ^ i u3 1 1 .7 5 11.4-3 13 .1 0 11.4-3 10.20 11.23 12.10 11.23 11.50 1 0 .2 7 10.4-3 11.00 11.00 1 0 .87 10.50 11-30 10 .87 12 .0 0 11 .23 12.00 11.30 10 .57 11 .37 3 1 .5 3 2 .5 3 6 .5 3 3 .0 28.0 3 2 .0 3 6 .5 3 0 .0 29-5 27.0 29.0 3 0 .0 3 0 .0 28.0 28.0 30.0 2 7 .5 33-0 2 9 .5 2 9 .5 2 7 .5 27.0 29.0 3 0 .4 26.9 27-433-2 28.7 3 6 .0 3 8 .5 3 3 .8 3 2 .3 3 2 .5 3 0 .3 3 3 .3 32-1 3 3-4 3 0 .7 29-4 33.0 33.0 31*3 29-5 27.3 35-0 3 0 .1 Mean Corpusc u le r Hemoglo­ b in uug 1 1 .4 9 .5 9 .8 11 -5 1 0 .5 1 2 .6 1 2.8 1 2 .6 1 2 .6 12.41 0 .9 1 2 .2 1 1 .8 13.0 1 1 .5 1 1 .1 1 3.0 12 .0 11-9 12.0 1 1 .2 13-7 1 1 .8 Mean Corpus­ c u la r Hemoglo­ b in Cone. 2 37.3 35-2 3 5 .9 3 4 .6 36.43 5.1 3 3 .2 3 7 .4 39-0 38 .0 3 6 .0 3 6 .7 3 6 .7 3 8 .8 3 7 .5 3 7 .7 3 9 .5 3 6 .4 38.1 4 0 .7 4 1 .1 39.1 3 9 .2 Serum T o ta l Albumin P r o te in gm i 6 .30 6 .3 5 6.58 gm io 4 .0 1 3 .7 1 3-98 —-----— 6 .1 9 5-9 6 6 .1 6 3 .8 4 3 -79 3-8 8 —-----— — 5.56 5-58 5-25 5-49 5.8 6 5-41 5-30 5-81 5-84 6 .4 8 6 .18 5-98 5.98 5.69 6 .2 8 — 3-71 — — 3-4 7 3-48 3-68 3-51 3-43 3 .6 1 3-48 3-78 3*75 3 .8 6 4 .0 1 3*46 3 .9 5 Calcium mg i> 1 1 .8 1 1.3 1 1 .4 1 2 .6 1 1 .5 1 1 .5 1 0 .8 1 1 .7 1 2 .2 1 1 .8 12 .0 12.0 1 2 .5 1 1 .1 1 1 .9 9 .5 1 0 .9 1 1 .5 10.3 1 2 .1 1 1 .5 1 1 .6 il.l P lasm a In o rg . Mag­ nesium P hosphoruB mg # mg 7 .3 6 7 .6 7 9 .3 3 3 .1 7 8 .0 6 8 .2 3 8 .0 6 7-58 8 .0 1 7 .81 7 .8 6 8.33 7 .6 2 7 .4 4 6 .5 4 6.07 4 .7 3 5-74 7-7 6 6 .9 8 7.23 6.83 6.13 2 .44 2.31 2.27 2.08 1.7 9 2 .5 5 1 .7 8 1.90 2.09 1 .6 2 2.43 2 .1 5 2.31 1 .9 2 1 .5 8 1.90 1 .6 8 2-75 2.07 1 .8 7 2.71 1.8 8 2 .6 6 i A scorbic Acid mg j> 0 .5 6 6 0.448 0.6 2 8 0.374 0 .4 06 0.423 0 .4 4 4 0.403 0 .5 4 5 0.519 0.407 0.413 0 .4 6 4 0 .4 4 9 0.400 0.370 0.3 5 1 0 .5 0 2 0.403 0 .5 6 8 0 .5 2 6 0 .6 1 2 0 .4 4 4 Appendix T able 9 Blood P ic t u r e o f Animal Humber C715— Group 2 Vthole Blood Bed Blood Hemo­ C e ll Count g lo ­ b in Age Red Blood C e ll Volume days M/mm3 gm £ £ 32 39 52 60 67 7^ 81 88 95 102 109 116 123 130 137 144 151 158 165 172 179 186 8 .9 5 9 .6 0 9 .8 0 9*20 1 1 .3 4 11 .76 12 .6 6 1 4 .29 13-95 15-43 16.00 1 8 .9 5 1 4 .8 4 15-15 18.60 1 6 .2 5 15-22 15-81 1 6 .3 7 15-40 17-50 15-87 1 3 .3 5 11.43 11.40 10 .6 3 11-93 12 .53 1 3 .1 7 1 3 .2 5 1 3 .2 5 1 5 .50 16.50 16.70 16.50 18.00 1 8 .7 5 18.10 18 .0 0 19.80 1 9 .77 1 8 .60 19-57 18.80 3 4 .0 3 0 .0 2 9 .5 2 9 .0 27.0 3 0 .0 3 3 .0 3 2 .0 30.0 3 6 .0 4 2 .0 4 1 -5 4 4 .0 4 6 .0 52.0 4 7 -5 4 4 .0 58-5 51-0 4 4 .0 4 5 .0 4 9 .0 Keen Corpus­ c u la r Volume u^ 3 8 .0 3 1 .3 3 0 .1 3 1 .5 2 3 .8 2 5 .5 26.1 2 2 .4 2 1 .5 23-3 26.3 21.9 2 9 .6 3 0 .4 28.0 2 9 .2 28.9 3 7 .0 3 1-2 2 8 .6 2 5 .7 3 0 .9 Meen Corpus­ c u la r Hemoglo­ b in uug Mean Corpus­ c u la r Hemoglo­ b in Cone. £ 1 4 .9 1 1 .9 1 1 .6 1 1 .6 1 0 .5 1 0 .7 10 .*1 9 .3 9 .5 10.0 10.3 8 .8 1 1 .1 11-9 1 0 .1 1 1 .1 1 1 .8 1 2 .5 1 2 .1 1 2 .1 1 1 .2 1 1 .8 39-3 38.1 3 8 .6 3 6 .7 4 4 .2 4 1 .8 39-9 4 1 .4 4 4 .2 43-1 39-3 4 0 .2 3 7 .5 39-1 3 6 .1 38.1 4 0 .9 33-8 3 8 .8 4 2 .3 4 3 -5 3 8 .4 Serum Albumin T o ta l P r o te in gm % gm £ ----- —---— --- — --- — ----- — ----- — —---— --- — ------- — --- — --- --- 7-54 6 .30 6 .94 7.*44 7-18 7-38 7-14 6.86 ----- ---6 .3 1 6.49 6.37 ---- — 13 3-99 2.77 4 .1 4 4 .1 2 4 .0 1 3-91 4 .0 4 ----------4 .1 4 4 .3 4 ———— Calcium mg £ 1 2 .5 1 2 .2 1 1 .5 1 2 .5 11-7 12 .3 1 0 .7 1 2 .6 1 1 .6 9*8 1 0 .1 1 2 .1 10.3 10-7 10 .6 11 .6 1 0 .5 9 -7 9-8 1 0 .1 9-6 1 0 .1 Plasm a In o rg . Mag­ nesium Phos­ p h o ru s mg £ 7 .8 6 6.13 6 .9 5 7 .1 9 5-41 6 .07 5-39 5-02 4 .5 5 6.32 7 .0 2 5-17 5-90 6 .2 8 5-79 4 .4 3 6 .1 9 6 .19 6 .2 8 6.87 5-74 6 .76 A scorbic Acid mg $ mg % 1-71 1 .6 8 1-75 2 .0 4 1.28 2.0 6 2-53 1 .8 3 1.6 7 1.9 8 3-63 2 .2 3 2.47 2 .6 5 2 .5 8 3-79 2.16 1.2 6 2.7 2 2 .11 2.58 2.00 0.367 0 .3 9 5 0.723 0 .*480 0.500 0 .4 9 7 0 .5 5 2 0.3 7 2 0.2 9 6 0 .6 4 2 0.5 7 6 0.563 0.5 8 4 0 .4 8 8 0.401 0 .2 7 9 0.596 0 .518 0 .4 8 2 0 .4 4 4 0.610 0.486 Appendix Table 10 Blood P ic tu r e o f Animal C?39— Group 2 Age Whole Blood Bed B locd Hemo- Bed Meen C e ll Count g lo - Blood C orpusb in C e ll c u la r Volume Volume days M/ mm'3 gm $ ~K u^ 35 42 1*9 56 63 70 77 81+ 91 97 106 111 118 125 132 139 li+6 153 159 167 y?h 181 188 196 203 209 216 223 9.30 8 .6 8 9 .2 2 1 0 .5 5 11 .59 11.69 13-30 13-60 15-12 li+.O 5 1 5 .9 9 1 4 .9 5 15-28 15*28 1 8 .5 7 19-53 18.33 1 8 .31 1 9 .6 2 1 8 .9 2 20.30 9 .7 0 1 0 .3 5 9 .9 0 11.93 12.60 12 .77 1 3 .2 5 12.83 12.53 1 2 .U5 1 2 .60 13.10 12.53 1 3 .9 5 1 4 .4 5 15.00 14.03 15-20 16.00 16.80 17.80 24.0 27.0 26.0 3 2 .5 3 4 .0 3 4 .5 3 5 .0 35-5 3 4 .5 3 2 .5 35-0 37.0 3 4 .5 4 2 .0 3 8 .0 4 2 .0 4 1 .5 4 8 .0 4 6 .5 5 0 .5 4 7 .0 2 5 .6 3 1 .1 2 8 .2 30.8 29.3 2 9 .5 26.3 2 6.1 2 2 .8 2 3 .1 2 1 .9 2 4 .7 22.6 2 7 .5 2 0 .5 2 1 .5 2 2 .6 2 6 .2 23.7 26-7 2 3 .2 — 19 .1 7 1 9 .7 7 1 8 .1 7 1 9 .2 7 1 5 .81 1 8 .1 2 — 17.10 17.90 17.80 18.10 17.60 1 8 .6 7 Mean Corpuj c u la r Hemoglo­ b in uug Mean Corpus­ c u la r Hemoglo­ b in Cnnc. i 1 0.4 11-9 1 0 .7 11.3 1 0 .9 1 0 .9 1 0.0 9 .4 8 .3 8 .9 7 .9 8 .8 8 .2 9 .1 7 .8 7 .7 7 .7 8 .3 8 .2 8 .9 8 .8 4 0 .4 38 .3 3 8 .1 3 6 .7 37.1 37.0 37-9 3 6.1 3 6 .3 38-3 3 6 .0 35-4 3 6 .3 3 3 -2 38.0 35-7 33-8 3 1 .7 3 4 .4 33.3 37-9 — ------ — -------- 4 7 .5 4 7 .5 4 5 .5 4 9 .0 4 8 .0 4 5 .5 24.8 24.0 25.0 2 5 .4 3 0 .4 25.1 8 .9 9 .1 9-8 9 .4 11.1 1 0 .3 36.0 3 7 .7 39.1 3 6 .9 3 6 .7 4 l .o Serum T o ta l Albumin P r o te in gm $ 5.60 5.49 6.40 5-48 5.93 6 .0 7 6 .07 0 .3 1 6 .0 4 5-95 6.30 6 .4 4 6-37 6 .6 5 6 .3 7 5 .9 5 6 .61 6 .16 6 .2 4 6 .6 6 6.31 6.89 6 .7 5 6 .4 4 7.23 6 .3 0 7 .2 8 gm f> 3-51 3*46 3 .9 6 3-57 3 .9 2 4 .0 9 3-93 4 .0 3 4 .0 4 3 .8 5 4 .1 7 4 .2 9 4 .2 1 — 4 .2 6 3 .8 2 4 .6 2 4 .1 4 2 . ?4 4 .2 1 3 .6 1 4 .2 0 3-70 4 .1 6 4 .63 3 .9 4 4 .5 9 Calcium Plasm a In o rg . MagF hosnesium c h o ru s mg £ mg $ mg $ 1 1 .4 1 2 .2 1 1 .8 12-7 1 0 .4 1 0 .7 1 1 .7 1 0 .7 1 1 .7 10.0 1 1 .0 1 1 .5 1 0 .1 12.0 11.8 9 .8 10.1 10.6 1 0.0 1 1 .3 10.0 1 0 .0 1 0 .5 10.3 10.8 11.3 9 .5 1 0 .4 6 .2 5 6.58 8 .0 6 6.83 7-86 4 .9 6 9-19 10.4 1 6 .98 8 .3 9 6 .5 1 1 1.57 7 .7 1 8 .8 7 6.83 7 .8 6 7 .0 6 5-63 6.51 6.91 4 .6 3 5*46 5-06 7 .3 1 4 .4 5 4 .8 4 5-53 3 .9 5 2 .1 5 2.21 2 .2 3 1-99 2 .57 1.53 2.99 2.20 2.07 1 .9 1 2.57 2.69 2.30 2.08 2-37 2 .47 2.01 1 .8 9 1 .9 2 1 .9 4 1-79 l.? 6 1 .8 7 1 .6 4 1 .5 6 2.07 1.80 2.58 A sco rb ic Acid ag 0.437 0 .5 4 4 0 .6 2 2 0 .6 8 2 0.623 0.583 0 .4 9 9 0-669 0.650 0.670 0.701 0 .6 9 2 0 .7 1 2 0.631 0 .6 7 5 0.5 6 4 0 .5 9 7 0 .5 1 9 0 .5 8 6 0 .563 0.503 0.4 1 4 0 .5 2 5 0 .4 7 4 0.371 0 .3 6 9 0-389 0.287 , co ro Appendix Table 11 Blood P i c tu r e o f Animal Humber C71+1--Group 2 Whole Blood Bed Blood Hemo­ C e ll Count g lo ­ b in 33 i+O 1+7 & 6l 68 75 81 90 95 102 109 116 123 130 137 li+3 151 158 165 172 180 187 193 200 207 Mean Corpus­ c u la r Volume Hf gm % % u? Mean Corpus­ c u la r Hemoglo­ b in uug 13*77 12.90 12.80 lh.l+4 15.60 12.83 15-57 1 8 .3 8 1 7 .0 6 1 8 .9 5 1 8 .2 8 1 8 .8 5 20.51 20.59 20.1+7 20.38 2 0 .0 5 2 0.59 20.03 1 9 .8 1 1 9 . so 2 0 .1 5 20.60 19-1+1 17*22 1 8 . 1+2 13 .77 13.50 13*35 13.70 13*77 1 3 .2 5 1 3 .7 7 lh -1 0 l h .0 3 11+.70 l h .80 1 5 .50 15*60 1 6.10 1 6 .50 1 6.90 16.80 17.30 1 8 . ho 18.50 1 9 .5 7 1 9 .5 7 1 9 .87 19 .3 0 19-50 1 9 .5 ^ i+o.o 29.0 2 8 .7 3 0 .5 29.1 2 6 .9 3 1 .2 2 5 .7 1 9 .9 2 5 .2 21+.8 2 3 .8 2 5 .2 2 2 .7 2 3 .6 21+.1+ 21+. 7 2 7 .7 2 5.7 27.0 2 7 .3 2 9 .0 26.0 2 7 .2 2 9 .6 3 1 .6 26.3 1 0 .0 1 0 .5 1 0 . 1+ 9 .5 8 .8 1 0 .3 8 .8 7 .7 8 .2 7 .8 8 .1 8 .2 7 .6 7 .8 8 .1 8 .1 8.1+ 8.1+ 9 .2 9 .3 9 .9 9 .7 9 .6 1 0 .2 1 1 .5 1 0 .6 Age day 8 Bad Blood C e ll Volume 3 7.0 3 9 .0 1+2 .0 1+2 .0 1+0 .0 1+0.0 3 6 .5 1+3 .0 1+7 .0 1+3 .5 1+7 .5 1*6 .5 1+8 .5 50.0 5 1 .5 5 5 .5 5 3.0 5i+.o 51+.0 5 7 .5 5 6 .5 56.0 5 7 .5 51+. 5 1+8 .5 Mean Corpus­ c u la r Hemoglo­ b in Cone. % 31+.I+ 3 6 .5 31+.2 3 2 .6 3 2 .8 3 3 .1 31+.1+ 3 3 .6 3 2 .6 3 1.3 31+.0 3 2 .6 3 3 .5 33*2 33.0 3 2 .8 3 0 .3 3 2 .6 3 h -l 3 ^ .3 31+.0 31+.6 3 5 .5 31+.1+ 36.3 1+0 .3 Serum Albumin T o ta l P r o te in gm i 7 .2 9 5 .88 6.21 5-91 6 .07 5.81 6 .1 6 5 . ?l+ 5.93 5.86 5* 98 6 .6 8 6 .21 5 .6 9 6.1+0 5.93 6 .0 2 6 .2 6 6 .23 6 .9 1 6 .8 9 6 . 51+ 6 . 1+1+ 6 . 11+ 6 .1+9 Calcium Plasm a In o rg . Mag­ nesium Phos­ p h o ru s A sco rb ic A cid gm ja mg rag # rag # rag $ l+.l+l 3 .3 7 3 .88 3 .8 6 1+.03 3 .8 1 1+.12 3 .7 9 1 1 .8 1 1 .5 1 0 .6 1 1 .9 1 1 .5 1 0 .1 1 1 .2 1 1.0 1 1 .1 1 1 .0 1 0 .6 1 1 .7 1 0 .8 11.1+ 9 .5 1 0 . 1+ 1 0 .1 ---— 1 1 .6 9 .6 9 .6 9 .3 1 0 .2 1 1 .5 9.1+ 1 0 . 1+ 6.83 7 .5 8 7*1+9 6 .9 5 8 .9 3 8 .0 6 8 .2 8 8.33 8 .6 1 9.h0 6 .6 5 8 .3 9 8 .0 6 9 .0 6 6 . 1+1+ 6 .5 8 6 .51 2.11 2.21 2.19 2.31+ 3.50 2 .3 8 2 . 21+ 2 . 1+7 3-02 3-02 2 .0 2 2 .91 2 .7 h 2 .0 5 2.1+3 1 .6 2 2 .1 5 1+.90 3 .69 l+ .ll 6.98 1+.07 5.56 1+.68 1+.12 1.73 1 .8 8 1 .9 0 0.508 0.5 2 7 0.6 9 8 0.5 5 0 0.573 0 .5 9 6 0.1+91 0 .5 7 9 0 .5 3 0 O .5I+6 0 - 611+ O .56I 0 .6 8 2 0 .6 0 5 0 .729 0 .5 1 1 O .63I+ 0.593 0.51+2 0.1+81+ 0.1+91+ 0 . 5I+I+ 0-513 0.531+ 0 . 1+59 0 . 1+68 3*98 l+.Oh 1+.13 l+ .ll 3 .9 2 1+.29 1+.08 3 .7 2 3 .9 6 1+.03 1+.37 l+.l+l 1+.21+ 1+.21+ -- — 2.61 1 .7 8 1.53 2.28 1.63 2.23 to v_o .Appendix S ab le 12 Blood P i c tu r e o f A n ita l Number C744— Group 2 Whole Blood Bad Blood HemoC ell Count> £ lo b ln Age Red Blood C ell Volume Mean Corpu&cu la r Volume days M/ram^ gm $ > u3 12 8*39 9 -0 5 8 .5 9 8 .9 4 9 -7 8 9 . to 1 1 .5 2 1 1 .7 8 13*98 1 4 .8 5 1 2 .6 7 1 5 .5 0 1 7 .5 0 1 0 .5 0 1 0 .5 7 9 .0 3 9 .7 0 1 0 .5 0 9 .3 3 9 . to 2 8 .0 2 7 .5 2 3 .0 2 6 .5 3 0 .0 2 5 .5 2 8 .0 2 5 .O 2 7 .5 3 3 -4 3 0 .4 2 6 .8 2 9 .6 3 0 .7 2 7 .1 24.3 2 1 .2 1 9 .7 2 0 .9 2 1 .3 2 0 .0 1 7 .1 1 8 .8 1 7 .9 1 6 .4 1 9 .5 1 7 .0 1 9 .7 1 7 .7 a .3 19 26 33 40 47 54 6o 69 74 81 88 95 102 109 16.98 166 1 7 .9 2 2 1 .3 8 1 9 .7 2 1 8 .2 6 2 0 .3 2 1 9 -7 6 1 9 .7 3 2 1 .^ 5 2 1 .2 4 172 179 186 2 1 .2 0 2 1 .8 8 116 122 130 137 144 151 159 a .9 5 8 .8 7 9 .5 5 1 0 .27 9 .2 0 1 0 .50 10.50 10.43 1 0 .6 3 1 0 .8 7 1 1 .5 0 1 1 .3 0 1 3 .7 0 1 2 .1 0 1 2 .8 3 1 4 .3 0 1 4 .5 5 1 4 .1 0 1 4 .6 3 15-50 3 1.0 2 7 .0 3 1 .0 3 0 .0 3 2 .0 3 2 .0 3 5 .0 3 8 .5 3 1 .0 4 1 .0 3 5 .0 4 2 .0 4 5 .5 4 6 .0 4 6 .0 4 6 .5 4 2 .5 2 1 .2 a .7 2 1.0 2 1 .9 1 9 .4 Mean Corpus­ cu la r Hemoglo­ bin uug 1 2 .5 1 1 .7 1 0 .5 1 0 .9 1 0-7 9 .9 8 .2 7 .5 6 .8 6 .9 7 .3 6 .8 6 .0 6 .1 5 .9 5 .1 5-3 6 .2 6 .6 6 .1 6 .5 6 .7 6 .9 6 .4 6 .9 7 .1 Mean Corpus­ cu lar Hemoglo­ b in Cone. Serum T otal Albumin P r o tein Calcium Plasma In org. Phos­ phorus i gm $ gm mg i mg £ 3 7 .5 5-08 5 .2 6 5 .3 0 5-78 5 .7 6 6 .0 5 5 .6 8 5 .9 6 6 .0 2 5-98 —6 .0 9 6 .5 6 5 -9 5 5-7 8 6 .1 3 3 .3 3 3 .4 0 3*46 3 -9 9 3 .7 7 1 1 .6 7 .7 6 8 .0 1 6 .2 2 7 .0 2 3 8 .4 3 9 -3 3 6 .6 3 5 -0 3 6 .6 3 3 -6 3 5 .5 3 4 .7 3 3 .1 3 4 .1 33-9 3 5-0 3 2 .6 3 3 .2 3 1 .1 2 9 .9 3 6 .5 3 3 .4 3 4 .6 3 0 .5 3 1 .4 3 1 .6 3 0 .7 31*5 3 6 .5 6 .2 6 5.61 6 .5 3 6 .56 6 .9 3 7 .2 4 6 .3 3 6 .4 8 6 .4 0 7 .1 4 3 .8 5 3 .8 5 3 .9 0 3 .8 6 3-8 9 — 3 .9 4 — 4 .0 1 3-90 4 .1 4 4 .2 1 3 .9 1 4 .0 3 3 .5 3 4.2.6 4 .3 6 3 .9 7 4 .1 6 2 .7 1 2 .9 6 1 1 .0 1 0 .8 1 0 .8 1 1 .5 1 1 .4 1 1 .3 1 1 .5 1 1 .3 1 1 .4 1 1 -5 1 1 .9 1 1-7 1 1 .7 1 0 .5 11.0 1 2 .4 1 2 .1 1 1 .0 1 0 .9 1 1 .2 1 3 .1 1 0 .1 1 1 .3 1 1 .1 1 0 .7 8.50 8 .7 4 7 .9 1 7 .9 6 9 .0 6 9 .9 2 7 .0 6 8 .9 3 8 .1 7 8 .0 1 8 .3 9 8 .3 9 9-19 7 .7 1 6 .7 2 4 .8 3 5 .2 3 7 .3 5 6 .6 2 6 .o 4 4 .7 0 3 .3 0 Mag­ nesium A scorbic Acid mg $ mg $ 2.36 0 .4 8 4 0 .3 6 8 0 .4 8 0 0 -3 6 4 0 .6 1 5 0 .5 5 5 0 .5 9 0 0 .6 8 5 0 .6 5 7 1 .6 5 2 .0 1 2 .1 4 2 .1 0 1 .8 9 1 .9 4 1 .9 0 2 .4 3 2 .4 8 2 .0 4 2 .1 5 1 .6 9 2 .2 6 2.18 1 .6 6 2 .7 2 1 .7 7 1 .9 4 2 .0 7 2 .8 3 2 .7 7 1 .8 8 2 .6 9 1 .5 4 2 .1 4 0.600 0.6 6 0 0 .5 7 7 0 .4 5 6 0 .4 9 9 0 .6 8 3 0 .5 0 3 0.6 1 0 0 .5 5 5 0.619 0 .5 0 0 0 .6 0 3 0 .5 7 5 0 .6 3 7 0 .5 2 6 0.451 0 .5 3 7 CD ST .Appendix T able 13 Blood P ic tu r e o f Animal llumber C711— Group 3 Whole Blood Red Blood Hemo­ C e ll Count g lo ­ b in Red Blood C o ll Volume Mean Corpus­ c u la r Volume days Yi/mn? gm 'j> £ u3 Meen Corpus­ c u la r Hemoglo­ b in uug 81 88 101 109 116 123 130 137 144 158 165 172 179 136 193 200 207 10-50 1 1 .9 5 1 5 .2 5 15.57 1 4 .3 4 1506 1 5 .23 15-85 15*30 1 5 .5 5 1 2 .1 5 1 1 .9 5 1 1 .1 0 10.90 8 .9 9 11.4-5 1 0 .6 2 10.87 1 1 .0 7 1 4 .33 13*60 14.43 14.90 16.00 15.60 15.90 16.80 1 3 .3 5 12.83 1 1 .6 7 1 3 .1 7 1 1 .5 5 13.60 13.77 27.0 3 0 .0 4 0 .5 4 0 .5 3 6 .0 3 6 .5 3 8.0 4 1 .0 3 7 .5 4 1 .5 3 4 .5 35-0 3 0 .0 33-0 29.0 34.0 37.0 25-7 25.I 2 6 .6 26.0 2 5 .1 2 3 .8 25.0 25.9 2 4 .5 26.7 2 8 .4 2 9.3 27.0 3 0 .3 3 2 .3 2 9 .7 3 4 .8 1 0 .4 9 .3 ' 9 .4 8 .7 1 0 .1 9 .7 1 0 .5 9 .8 1 0 .4 1 0 .8 11.0 1 0 .7 1 0 .5 1 2 .1 13-2 11.9 13.0 Age Mean Corpus­ c u la r Hemoglo­ b in Cone£ 4 0.3 3 6 .9 3 5 .4 33-6 4 0 .1 4 0 .8 4 2 .1 38.0 4 2 .4 4 0 .5 3 8 .7 3 6 .7 3 3 .9 3 9 .9 4 0 .9 4 0 .0 3 7 .2 Serum Albumin T o ta l P r o te in gm $ —— —— ---- — ---------------6 .1 9 6 .8 9 6.21 6.70 6 .2 3 7 .2 3 6.26 6 .5 4 ------ ------------------------ — ----------3-80 4 .1 6 3 .8 6 3 .9 6 3-78 3*79 -----4 .1 3 —---- Calcium mg $ 11-7 1 2 .0 11-3 11-5 9 .3 1 0 .5 9 .5 11 -5 1 1-5 11-7 ' 1 1 -5 1 1 .2 1 1 -5 1 3 .1 11.8 1 0 .1 1 0 .0 Plasm a In o rg . Mag­ nesium Phos­ p h o ru s A scorbic Acid mg

mg $ 6 .0 2 6 .2 4 5 .96 6.21 5 .9 5 6 .5 8 6 .6 6 6 .59 —---— 3 .0 4 3*28 3 .3 7 3 .37 3 .3 3 3 .6 6 3 .6 6 3 -9 4 1 0 .9 11*5 1 2 .3 1 1 -5 1 1 .5 1 1 .5 13.0 1 1 .1 1 1 .4 1 0 .7 1 1 .6 1 0 .5 1 0 .7 9 .6 9 .5 10-7 1 1 .5 1 0 .6 10.c 1 0 .5 1 1 .2 1 0 .5 9 .3 1 0 .7 1 0 .6 6 .19 6.69 7-31 7 .4 4 7 .62 6.48 6.79 8 .7 4 7.71 9 .4 7 8 .0 6 8 .1 7 8 .3 9 6.0 1 4 .6 6 6 .8 7 7*02 6.13 7 .0 2 6 .1 9 8 .3 3 7 .1 9 6 .1 3 4 .5 1 6 .0 4 T O'** X •w 0.228 0.3 9 9 0 .2 5 5 0.469 0 .4 5 6 0 .6 2 6 0 .4 6 8 0.658 0 .5 0 3 0 .5 2 2 0.2 6 7 0 .5 5 5 0.5 2 7 0.419 0 .3 3 5 0 .3 4 5 0 .4 j4 0*420 0.4 3 7 0 .5 4 4 0 .4 1 4 0.3 6 0 0 .4 4 6 0.550 0.482 6 .2 6 6.30 6 .6 3 6 .1 9 -----—---— 6 .7 4 6 .83 6 .7 4 7 .0 7 6 .7 4 6 .6 5 6.93 6 .7 9 -----4 .1 8 3 .8 6 -----3 .9 5 3 .6 4 ----------3-8 5 3-94 3 .81 4 .1 8 3 .8 2 3 .8 2 4 .0 3 3*86 2 .06 1.S0 1 .9 5 1 .80 2.53 3*89 1 .8 3 1 .9 6 2 .47 2.93 2.28 2.31 2 .4 3 2 .21 2.69 2 .1 5 2.53 2.21 1 .9 7 2 .4 4 1.93 2.23 1 .8 8 2.38 Appendix T able 14 - Cont inued Blood P i c tu r e o f Animal Number C725—Group 3 Whole Blood Bed Blood Hemo­ C e ll Count g lo ­ b in Age days M/mm^ g ° 7= 207 214 220 229 234 2 kl 248 255 262 269 276 282 290 297 304 311 319 326 332 339 346 1 4 .2 2 13*73 1 4 .17 1 4 .9 2 1 6 .51 13*77 13*03 13*63 15*56 16.13 1 4 .2 4 14 .0 8 13-91 13-38 14.10 13*02 12.53 1 1 .3 8 11*55 11.30 11-37 18.00 17*00 17-35 17-80 17-50 18.10 18.30 17-90 1 8 .7 2 19-10 18.40 1 8 .10 17*50 17*50 17*40 17*00 16.60 16.30 15.60 15-40 15.50 Bed Blood C e ll Volume u3 Kean Corpus­ c u la r Hemoglo­ b in uug Mean Corpus­ c u la r Hemoglo­ b in Cone. *4 7= 3 2 ,0 3 0 .2 3 3 .5 3 2 .8 2 8 .8 35-6 3 7 .6 3 2 .3 3 0 .5 3 3 .2 3 5 .1 3 5 -5 3 7 .0 3 2 .1 3 1-6 3 2 .3 2 8.7 3 3 .8 3 4 .2 29.6 29.9 1 2 .7 1 2 .4 1 2 .6 1 1 .9 1 0 .6 13-1 1 4 .0 1 3 .1 1 2.0 1 1 .8 1 2 .9 1 2 .9 1 2 .6 1 2 .6 1 2.3 13-1 1 3 .2 14.3 1 3 .5 1 3.6 1 3 .6 39-6 4 1 .0 3 7 .6 3 6 .3 3 6 .8 3 6 .9 3 7 .3 4 0 .7 3 9 .4 3 5 .7 3 6 .8 3 6 .2 3 4 .0 39-3 3 9 .1 4 0 -5 4 6 .1 4 2 .3 3 9 .5 4 6 .0 4 5 .6 Kean Corpus­ c u la r Volume "" ** 73 4 5 .5 4 1 .5 4 7 .5 4 9 .0 4 7 .5 4 9 .0 4 9 .0 4 4 .0 4 7 .5 5 3 .5 50.0 50.0 5 1 .5 4 4 .5 4 4 .5 4 2 .0 3 6 .0 3 8 .5 3 9 .5 3 3 -5 3 4 .0 J l k Serum T o ta l A ltu n in P r o te in Calcium P lasm a In o rg . Mag­ nesium P hos­ p h o ru s A sco rb ic Acid gm % gm # mg $ mg % mg % mg i 7.23 7 .0 7 7 .3 1 6 .9 4 6 .5 3 6 .8 6 6 .7 5 7-01 6-53 6 .2 6 6 .7 7 6 .37 6.37 6 .3 8 7 .2 6 7-13 7 .1 4 6 .1 4 6.93 6 .5 9 7 .0 7 4 .0 5 4 .1 0 4 .2 2 3 .00 3 .8 6 4 .1 8 3 .8 5 —---3 .9 0 3*75 -----3 .60 3.73 3.53 3 .6 1 3 .7 4 3-98 2 .7 2 3 -8 4 3 .6 4 3 .7 3 9-1 10-3 10-7 9 .9 1 0.0 9 .5 1 0 .5 9 .5 9 .6 8 .8 1 0 .7 1 0 .1 9 .7 9 .5 1 0 .8 1 0.0 1 1 .9 11.4 1 2 .4 1 0 .5 1 1 .1 6 .9 5 7.81 6 .9 1 7.71 7 .4 4 6 .0 1 8.23 7 .0 2 6 .9 8 6 .6 2 5.56 5.90 6.58 5-84 3 .6 7 4 .1 5 5-56 4 .9 4 5.27 5-81 5.81 2.07 2 .0 6 2.11 2 .1 5 2.18 1 .90 2.43 2.11 1 .9 4 2 .3 3 2.58 2.50 2.10 1 .7 4 1 .7 9 2.83 2 .7 4 2 .3 2 3-04 1 .7 6 2 .5 5 0 .4 5 5 0 .3 5 8 0 .4 5 5 0 .5 1 5 0-546 0 .4 8 2 0 .5 6 1 0.613 0.4 9 1 0 .5 0 2 0 .5 3 5 0 .4 6 6 0 .3 4 4 0.480 0.343 0 .2 8 5 0 .4 4 2 0.451 0 .3 8 5 0.420 0 .3 1 9 A ppendix T able 15 B lood P ic tu r e o f Animal Humber C730— Group 3 Whole Blood Bed Blood Hemo­ C e ll Count g lo ­ b in Age day s Uf mm3 ng $ 37 44 51 58 65 72 79 86 93 100 107 114 121 128 135 142 149 156 163 170 177 184 191 198 205 212 11.11 10.81 1 1 .1 5 9 .53 -------1 0 .2 5 10.40 14.90 ------ 1 4 .7 4 15.20 1 4 .0 2 1 4 .99 1503 15-95 ---- —15.60 1 6 .1 4 1 7 .1 2 16.80 16.70 1 6 .8 5 1 6 .3 2 1 6 .3 4 18.03 15-38 10.13 1 0 .13 13.70 12.10 13.60 12.53 14.10 15.90 —-----15.60 17.00 1 7 .50 18.30 17-90 18.40 19.26 19.20 18.80 19-54 18.80 1 9 .6 6 19-54 19.40 18.50 1 9 .50 18 .60 Bed Blood C e ll Volume Mean Corpus­ c u la r Volume * 2 7 .5 28.0 4 5 .0 3 5 .0 3 8 .0 3 4 .5 3 5 -5 4 5 .0 -----4 0 .0 4 9 .0 4 7 .0 52.0 4 8 .0 5 0 .5 5 4 .5 5 3 .5 4 9 .0 5 3 .5 52.0 59.0 5 8.0 57.0 5 3 .5 5 3 .5 52.0 u3 Mean Corpus­ c u la r Hemoglo­ b in uug Mean Corpus­ c u la r Hemoglo­ b in Cone. 2 2 4 .8 2 5 .9 4 0 .4 3 6 .7 -----33 -7 3 4 .1 3 0 .2 9 -1 9 .4 1 2 .3 1 2 .7 ---- 1 2 .2 1 3 .6 10-7 2 7 .1 3 2 .2 3 3 .5 3 4 .7 31.3 3 1 .7 —---— 3 4 .3 3 0 .4 3 1 .3 3 1 .0 3 5 .3 3 4 .4 3 4 .9 3 2 .7 2 9 .7 33*8 1 0 .6 1 1 .2 1 2 .5 1 2 .2 1 1 .7 1 1 .5 —~ — 1 2 .3 1 1 .6 1 1 .4 1 1 .2 1 1 .8 1 1 .6 1 1 .9 1 1.3 10 .8 1 2 .1 3 6.8 3 6 .2 3 0 .4 3 4 .6 3 5 .8 36 .3 3 9 .7 35.3 ——— 39.0 3 4 .7 3 7 .2 3 5 .2 37-3 3 6 .4 35-3 35-9 3 8 .4 3 6 .5 3 6 .2 33-3 3 3 .7 3 4 .0 3 4 .6 3 6 .4 35-8 Serum T o ta l Albumin P r o te in Calcium In o rg . Phos­ p h o ru s Plasm a Mag­ nesium A sco rb ic Acid gm $ gm f> mg i mg j> mg $ mg $ 6 .0 9 5 .96 6 .1 3 5 .6 5 6 .0 5 6 .0 7 5-98 ---- — -----5 .84 6 .0 4 ---- — 6.66 6.33 3 .6 6 3*71 .3.83 3 .4 6 3 .4 6 3 .5 9 3 .7 6 ---- -----3 .8 1 3 .9 1 -----4 .2 1 4 .1 4 ---— 6 .6 8 6.63 6.79 6.9 1 7-18 6.83 7 .1 2 6 .7 2 6 .8 9 7 .0 5 3 -9 4 4 .3 2 4 .3 4 4 .2 1 4 .7 9 4 .1 9 4 .6 4 4 .3 8 4 .8 4 4 .6 6 1 2 .1 1 2 .5 1 1 .6 ll.l 1 1 .5 1 1 .7 1 1 .2 1 0 .7 1 1 .5 1 1 .9 1 0 .4 10 .0 1 0 .4 1 0 .8 1 0 .4 1 0 .0 1 0 .5 9 .6 1 0 .5 1 0 .5 1 1 .9 9 .3 1 0 .7 1 0-5 1 0 .9 1 0 .0 8 .4 5 7 .5 8 7.23 7 .0 2 6 .9 5 7 .9 1 7 .7 1 7 .7 1 8.33 6 .4 5 7 .6 2 7 .3 5 6.98 5.53 5-53 7 .4 9 6.10 7 .0 2 7 .7 1 7 .0 2 8.1 7 7 .6 7 4 .8 3 7 .8 1 5-63 7 .9 6 2 .3 7 2.33 2 .2 5 1 .90 2.29 2.5 5 2.53 2.4 9 1 .5 3 2.49 2.02 1 .9 0 2.21 2.20 2-53 1 .80 2.51 2 .16 2.20 1 .9 4 1 .9 4 2 .46 2.0 2 2.78 2 .47 2.49 0-391 0 .4 9 1 0 .4 7 7 0 .4 7 2 0.273 0-530 0.6 8 9 0 .6 5 6 0 .5 8 5 0 .5 0 7 0 .6 0 2 0 .6 7 1 0 .483 0 .4 0 1 0 .537 0 .4 9 8 0.3 8 0 0 .3 9 8 0.4 9 8 0.6 4 6 0 .2 4 8 0 .5 3 1 0-583 0 .4 7 4 0.471 0.3 2 7 Appendix Table 16 Blood P i c t u r e o f Animal Number 071*1— Group 4 Whole Blood Red Blood Hemo­ C e ll Count g lo ­ b in Age days M/mm^ gm i 46 3 66 74 81 88 95 102 109 116 123 130 137 144 151 158 165 172 179 193 200 207 214 221 228 9 .6 5 10.40 1 0 .2 5 1 0 .2 7 1 0 .7 5 1 0 .5 2 IO .38 10.80 9 .6 0 10.*40 1 0 .7 5 1 1 .3 4 10.90 11.70 1 4 .4 5 1 4 .2 5 1 4 .7 4 1 6.10 -------1 7 .3 0 -------17 .67 1 6 .5 5 1 7 .00 1 9 .6 5 1 1 .6 7 11.00 11 .40 1 1 .30 1 2 .4 5 10.93 12 .7 7 11-75 1 1 .6 0 1 2 .8 3 13.70 12 .7 7 1 3 .7 7 13.70 17.10 15-80 1 6 .50 16.80 -------1 9 .57 1 9 .6 7 19.50 19-70 19.50 2 1 .7 4 Red Blood C e ll Volume Mean Corpus­ c u la r Volume $ 2 9 .0 2 9 .5 3 2 .0 3 4 .0 3 2 .0 2 7 .5 3 1.0 29.0 2 9 .5 3 5 -5 3 7-5 3 5 .5 3 8 .0 37.0 51.0 4 2 .5 4 7 .0 4 9 .0 -----53.0 5 7 .5 5 6 .0 58.0 5 6 .5 6 3 .O u^ Mean Corpus­ c u la r Hemoglo­ b in uug Mean Corpus­ c u la r Hemoglo­ b in Cone. 2 3 0 .1 2 8 .4 3 1 .2 3 3 .1 29.8 2 6 .1 2 9.9 26.9 30-7 3 4 .1 3 4 .9 31.3 3 4 .9 3 1 .6 35-3 2 9 .8 3 1 .9 3 0 .4 - —3 0 .6 -----31-7 3 5 .0 3 3 -2 3 2 .1 1 2 .1 1 0 .6 1 1 .1 1 1 .0 1 1 .6 1 0 .4 1 2.3 1 0 .9 1 2 .1 12.3 1 2 .7 11.3 12 .6 1 1 .7 1 1 .8 1 1 .1 1 1 .2 1 0 .4 -----11.3 -----1 1 .0 1 1 .9 1 1 .5 1 1 .1 4 0 .2 3 7 .3 35-6 33-2 3 8 .9 3 9 .7 4 1 .2 4 0 .5 39-3 3 6 .1 3 6 .5 3 6 .0 3 6 .2 37-0 3 3 .5 3 7 .2 3 5 .1 3 4 .3 -----3 6 .9 3 4 .2 3 4 .8 3 4 .0 3 4 .5 3 4 .5 Serum Albumin T o ta l P r o te in gm £ gm i> — —— ——— ---— — —— ---- — — — -----6.07 6.30 6.23 6 .2 8 6 .4 1 6 .2 4 7-21 7 .4 9 6.93 — -----6 .5 9 7.0 0 7 .1 2 6 .6 6 ---- — —---— ---- — ------ —---— ---- ---- — -----3 .3 9 3-6 8 3-68 3 .7 3 3 .2 9 3 -36 3 .7 9 3 .6 2 3*53 -----4 .0 3 ——--4 .0 9 4 .0 5 Calcium P lasm a In o rg . Mag­ nesium Phos­ p h o ru s A sco rb ic Acid mg mg $ mg $ mg $ 1 1 .7 1 0 .5 1 0 .2 1 1 .7 1 1 .5 1 1 .4 1 1 .0 1 1.0 9 .5 9 .6 1 1 .5 1 1 .5 10.3 1 1 .5 1 0 .1 1 1 .2 10.3 9 .5 9 .8 8 .7 9 -5 8 .4 8 .6 9 .0 8 .3 7 .2 7 7 .4 4 1 0.08 7 .9 6 6 .6 5 6.5 1 7 .4 4 8 .6 2 8 .8 1 8 .4 5 10.41 7 .3 5 7 .6 2 7 .5 8 8 .0 1 6.83 8 .3 3 7 .2 7 7-3 5 7 .4 9 6 .6 5 5.27 5.79 5.37 6 .5 8 1 .1 4 1 .1 6 1 .6 1 1 .5 6 1 .6 7 1 .6 2 2.31 l.? 4 1 .9 9 1.8 0 2.66 1 .8 6 1 .9 8 1 .7 9 2 .77 3.80 1.90 1-58 2 .1 5 2 .8 8 2.44 2 .5 8 3 .1 2 3 .22 2.43 0 .2 1 9 0.323 0 .5 6 2 0 .4 3 4 0.4 0 7 0 .3 3 6 0.5 1 1 0 .4 2 2 0.4 3 9 0 .4 3 9 0.4 3 2 0 .2 7 2 0.576 0 .4 0 8 0.393 0 .468 0 .493 0.440 0 .4 8 2 0.6 1 0 0 .3 1 2 0.238 0 .4 3 4 0 .3 9 4 0.553 00 'O Appendix T able 17 Blood P i c t u r e o f Animal Humber C726— Group 4 Whole Blood Red Blood Hemo­ C e ll Count g lo ­ b in Age $ days M/mm^ gm 32 39 46 53 60 67 74 81 88 95 102 109 116 123 130 137 144 151 158 165 172 179 186 193 199 1 1 .8 5 1 0 .8 2 11-70 1 0 .0 7 11.00 12.00 1 1 .8 2 1 0 .1 7 1 0 .8 5 1 1 .1 2 9 .0 5 11.20 11.00 ----- — 10 .11 10 .5 9 1 1 .0 2 9 .5 0 9 .5 7 12.30 11.13 1 0 .81 1 1 .1 7 10.13 1 0 .6 7 1 2 .90 12.83 1 2 .77 11.50 1 2 .4 5 1 2 .3 5 12.10 U .60 11.50 11.60 9 .7 0 1 1 .1 5 11.00 U .6 0 12.00 1 2 .2 0 11.60 10-50 1 1 .3 7 1 3 .3 5 1 2 .8 3 12.53 12.00 12.83 14.10 Red Blood C e ll Volume Mean Corpus­ c u la r Volume i 3 9 .0 3 7 .0 3 5 .0 3 4 .5 3 6 .5 3 7 .0 3 6 .5 34.0 3 2 .0 3 4 .0 29.0 3 0 .0 3 1 .0 3 0 .0 3 2 .0 2 9 .0 29.0 2 9.0 3 3 .5 3 9 .0 3 5 .5 35*0 3 2 .0 35-0 3 7 .0 u-> Mean C orousc u la r Hemoglo­ b in uug Mean Corpus­ c u la r Hemoglo­ b i n Cone* "T " 3 2 .9 3 4 .2 2 9 .9 3 4 .3 3 3 .2 3 0 .8 3 0 .9 3 3 -4 2 9 .5 3 0 .6 3 2 .0 2 6 .8 2 8 .2 -----31*7 27*4 26.3 3 0 .5 35-0 31-7 3 1 .9 3 2 .4 2 8 .6 3 4 .6 3 4 .7 1 0 .9 1 1 .9 1 0 .9 1 1 .4 11.3 1 0 .3 1 0 .2 1 1 .4 1 0 .6 1 0 .4 1 0 .7 1 0 .0 1 0.0 ---- 1 1 .9 1 1 .5 1 0 .5 1 1.1 1 1 .9 11.3 1 1 .5 1 1 .6 1 0 .7 1 2 .7 1 3 .2 3 3 .1 3 4 .7 3 6 .5 33.3 3 4 .1 3 3 .4 33-2 3 4 .1 35-9 3 4 .1 3 3 .4 3 7 .2 3 5 .5 3 8 .7 3 7 .5 4 2 .1 4 0 .0 3 6 .2 33-9 3 5 .5 3 6 .1 3 5 .3 3 7 .5 3 6 .7 3 8 .1 Serum T o ta l Albumin P r o te in Calcium $ gm % mg 5*51 5.83 6 .7 5 7 .93 8 .6 4 —---- 2 .91 2 .83 1 .8 6 2 .5 4 2 .3 9 9 .3 9 9-01 8 .2 8 8 .6 4 8 .5 6 ---—— -----7 .2 7 7 .8 2 7 .07 7 .1 4 6.89 7 .0 5 6 .8 4 7 .1 2 7 .2 8 7.10 6 .8 9 2.56 2.48 2.50 2 .4 6 2 .4 6 —— ---—— 2.6 8 3 .0 6 3 .1 1 3 .2 6 3 .4 4 2.03 3 .7 8 3-72 3 .8 1 3 -81 3 .6 1 8 .7 ' 9 .1 1 0 .5 U .7 1 0 .7 1 0 .8 1 0 .7 1 0 .9 9 .5 7 .0 1 0 .4 1 1 .7 1 1 .6 1 1 .5 1 1 .3 1 0 .8 1 1 .1 1 1 .9 1 2.0 1 0 .7 1 0 .5 1 1 .2 1 0 .7 1 1 .2 9 .9 gm % Plasm a In o rg . MagP h o snesium ■ohorus mg % 4 .8 4 4 .0 9 4 .2 6 5-30 4 .9 6 6 .3 5 7.23 5-90 4 .3 6 6 .3 8 6 .6 2 4 .9 6 6 .6 5 5-58 7 .0 2 9 .2 6 8 .1 7 8 .23 9 .8 4 7 .8 1 6 .4 8 8 .0 6 8.53 7 .9 1 8 .7 4 mg j> 1 .2 6 1 .5 7 1 .9 7 3 .5 5 1 .5 3 2.33 1.70 2.67 1 .0 8 1 .5 6 1 .6 4 1 .9 6 2 .1 5 1 .8 3 2 .3 6 1 .7 5 1 .7 0 2.08 2.37 2.08 1 .8 1 2 .21 1*95 1 .83 1.63 A sco rb ic A cid mg $ 0.263 0 .2 3 7 0.176 0 .1 6 6 0 .1 9 7 0 .1 7 5 0.2 2 1 0-381 0 .2 5 2 0.193 0 .2 0 5 0 .3 1 8 0 .4 0 2 0.3 8 6 0 .2 8 3 0.491 0 . 291* 0 .4 2 2 0 .3 6 8 0.4 8 0 0.4 9 1 0.440 0-412 0.350 0 .3 8 5 Appendix T able 17 - C ontinued Blood P i c tu r e o f Animal Number 0726—Group 1* Whole Blood Red Blood Hemo­ C e ll Count g lo ­ b in Age day s M/mnr^ gm # 208 213 220 227 23** 21*1 21*8 255 261 269 276 283 290 298 305 311 318 325 13*68 12.01 12*97 13*28 13*22 13*1*0 1 4 . 38 13*83 1 3 .22 li*. 26 15-1^ 13.1*5 1 1 .6 6 1 1 .7 6 1 1.56 U . 91* 1 2 .2 5 1 1 .3 1 11*. 90 1 5 . CO 1 6 .00 17.50 17.10 17 .5 0 18.30 18.00 17.10 16.90 1 6.70 16.30 16.00 15*00 11*. 55 li*. 63 16.10 11*. 10 Red Blood C e ll Volume Keen Corpus­ c u la r Volume I Ur* 1*2.0 1*0.5 1*6.0 1*8.0 1*6.0 1*7-5 50.0 51.0 1*9-5 1*8.0 1*2.5 1*2.0 1*1.0 3 2 .5 3 8 .5 3 8.0 31*.0 3 3 .0 3 0 .7 3 3 .7 35*5 3 6 .1 3l*.8 35-1* 31**8 3 6 .9 37-1* 33-7 2 7 .5 3 1 .2 3 5 .2 2 7 .6 3 3 .3 3 1 .8 2 7 .8 2 9 .2 Mean Corpus­ c u la r Hemoglo­ b in uug 1 0 .9 1 2 .5 1 2.3 1 3 .2 1 2 .9 1 3 .1 1 2 .7 13.0 1 2.9 1 1 .9 1 0 .8 1 2.1 1 3 .7 1 2 .8 1 2 .6 1 2 .3 1 3 .1 1 2 .5 Meen Corpus­ c u la r Hemoglo­ b in Cone. Serum Albumin T o ta l P r o te in Calcium Plasm a In o rg . Mag­ nesium F h o sp h o ru s i gm % gm $ mg i mg % mg $> 3 5 .5 37-0 31*.8 3 6 .5 3 7 .2 36.8 3 6 .6 35-3 31**5 35-2 39-3 3 8 .8 3 9.0 1*6.2 37*8 3 8 .5 1*7.1* 1*2.7 7.21* 6 .8 9 6 .9 6 6 .8 8 7.38 6 .6 5 7.09 6 .9 1 6 . 71* 6 .6 5 6-70 6 .7 2 7 .31 7 .1 6 6 .5 9 6 .7 2 6.63 6.77 3-38 2.91* 3 .7 2 3 .6 8 -----3*72 3 .8 6 2 .2 8 3 .6 8 3*91 3 .5 6 3.91* 3-9 5 3 .9 9 3 .5 8 3 .7 2 3-59 3-73 1 0 .0 10.3 9 .8 1 0 .6 9 .9 9 .0 9 .0 9.C 1 0 .1 1 0 .5 9 .0 1 0 .1 1 1 .9 1 2 .5 1 0 .6 1 1 .6 1 0 .3 ll.l 8 .3 9 8 .71* 7 .2 7 8 .8 7 7.8 1 7-1*9 6 .83 5.90 6 . 1*8 5.79 i* .6 l i*.81 i*.9i* 6.28 6 .6 5 5 .9 5 5.79 1**77 2.06 1 .8 5 1 .9 7 2.1*1* 1 .6 6 2 .07 2.21 1 .9 8 2.08 1 .7 3 1 .6 1 1.91* 2 . 7I* 2.53 1 .8 7 2.21* 1.61* 2.1*7 A scorbic A cid mg £ 0.530 0.1*99 0.1*1*3 0.1*71* 0.582 0.1*21* 0 .5 0 2 0.1*1*0 0.511* 0-381* 0 .1*^1 0.1*11* 0 .5 2 5 o .m 0 .1*71* 0 .1*32 0.1*75 0.1*83 1 n o LJ , Appendix T able 18 Blood P i c tu r e o f M i j n e l Number A73— Group 5 Whole Blood Bed Blood HemoC e ll Count gLobin Age days 371 385 399 1*27 429 436 1443 450 457 464 471 477 484 492 498 512 519 526 533 540 M/nnn3 ------------— 10.13 9 .0 5 8 .5 0 1 2 .7 5 9 .1 8 9 .1 0 7 .9 5 7 .5 5 7 .9 2 9 .5 3 8 .0 5 8 .2 0 9 .1 5 1 0 .3 2 7 .2 4 708 8 .2 8 go 2 1 1 .1 5 10 .0 7 9 .5 5 12.00 10 .9 3 1 1 .8 5 11 .93 11.60 1 1 .23 1 0 .2 7 1 0 .3 5 10-67 10.63 10.00 10.87 15*00 12.10 9 .7 7 10.50 11.00 Bed Blood C e ll V ol. i 2 6 .5 25*5 24.0 3 1 .0 26.0 2 9 .5 29.0 26.0 27.0 24.0 2 4 .5 26.0 26-0 2 4 .5 2 8 .5 3 0 .5 3 2 .5 23.0 25.O 25.0 Mean Corpus­ c u la r Volume u3 Mean Corpusc u le r Hemog lo b in uug -----------— --------- ------------ 3 0 .6 2 8 .7 3 4 .7 2 2.7 28.3 2 9 .7 3 0 .2 3 2 .5 3 2 .8 27.3 3 0 .4 3 4 .8 33*3 31*5 3 1 .8 3 3-9 3 0 .2 1 1 .8 1 2 .1 13-9 9 .4 1 2 .6 1 2 .3 1 2 .9 13*7 13*5 1 1 .2 1 2 .4 13-3 1 6 .4 11-7 1 3 .5 1 4 .2 13-3 Mean C orpusc u l s r Hemog lo b in Cone. C a lcium Plasm a In o rg . P h o s- Magnep h o ru s sium A scorbic Acid i mg % mg $ mg $ 4 2 .1 39*5 40 .0 3 8 .7 4 2 .0 4 0 -2 4 1 .1 4 4 .6 4 1 .6 4 2 .8 4 2 .2 4 1 .0 4 0 .9 4 0 .8 3 8 .1 4 9 .2 3 7 .2 4 2 .5 42 .0 4 4 .0 11.3 1 1 .1 11*5 1 2 .5 5.4 6 6 .2 8 5-81 7 .3 5 1 .8 8 2.18 1 .7 6 2.00 0 .646 0.390 0-504 — ------------------- mg £ - - _ - — ------------ ------------ ------------ ------------ --------------- 11.3 6 .4 1 2 .7 4 O.528 — — ------------ --------------- ------------ --------------- — — - ------------ ------------ ------------ ------------ ------------ ---------------- —— ------------ --------- --- --------------- 0.4 1 2 — 1 1 .7 6 .0 7 1 .9 6 ------------ —— ------------ ------------ — 1 1 .7 1 2 .2 1 0 .8 1 1 .1 1 1.4 1 1 .6 1 2 .3 6 .5 4 6 .5 1 6 .7 9 6 .6 5 5- 48 5-81 8.3 9 2 .0 2 1 .7 8 1*57 2.19 2 .30 2 .2 4 1 .8 6 --------------- -- ------------- 0 .4 0 4 0 .4 9 5 vO r\j Appendix T able 19 Blood P i c t u r e o f Animal Humber A75— Group 5 W iole Blood Bed Blood Hemo­ C e ll Count g lo b in Ace day 8 323 337 351 379 381 388 395 402 409 416 423 429 436 444 450 464 471 478 485 492 M/mm^ gm £ Bed Blood C e ll V ol. i TP Mean Corpus­ c u la r Hemo­ g lo b in i , ,__ Men C ornusc u la r Hemo­ g lo b in Cone. uug C al­ cium Plasm a In o rg . P h o s- Magnep horu8 sium mg £ mg £ £> 1 1 .1 11.0 1 2 .9 1 0 .7 6 .7 6 6 .5 8 5-81 8 .8 1 1 .9 0 1 .98 2 .1 5 2 .46 0.5 9 1 0 .4 3 8 0 .3 6 6 3 8 .5 2 9.9 3 8 .4 14.14 1 2 .6 1 6 .2 ---------- —---------- —------- ---------- 1 2 .6 8 .5 0 2.37 25.0 2 7 .0 2 4 .5 1 9 .0 23.0 2 2.0 3 5 -5 3 0 .0 2 8 .5 3 3 -5 29.0 3 2 .5 3 2 .0 2 8 .6 37.0 3 2 .5 3 7 .6 33-2 3 2 .2 4 4 .3 3 5 .0 3 8 .8 39-5 3 3 -5 39 -5 34 .6 12-5 15-5 13-9 1 6 .2 13-3 1 4.7 1 7-2 1 3 .2 1 4 .9 1 5.0 1 4 .6 1 6 .7 15-4 43-7 4 1 .9 4 2 .9 43-2 4 0 .0 4 5 .8 38 .8 3 7 .7 3 8 .4 3 8 .1 4 3 .6 4 2 .2 4 4 .4 —------- ---------- ---------- ---------- _ — ---------- ---------- ---------- ---------- —-------- ---------- ---------- 1 0 .79 1 0.70 7 .8 1 2 6 .0 23-0 26.0 4 1 .5 3 2 .0 3 0 .0 ------------- ------------- 8 .7 3 7 .3 0 7 .5 5 5-05 6.93 6.83 8 .0 2 8 .5 6 7 .3 4 8 .4 9 8 .6 5 8 .2 2 9*25 1 0 .9 3 11.30 10.50 8 .2 0 9 .2 0 1 0 .0 7 13-77 11.30 1 0 .93 12 .7 7 1 2 .6 5 13-70 1 4 .2 0 -----— . —-----— -----—— A scorbic Acid mg £ 4 4 .6 4 1 .2 4 3 .5 37.3 4 2 .0 4 2 .2 11.60 9*47 11.30 15-50 13.43 1 2 .6 5 ---------- Mean Corpus­ c u la r Volume ---------- ---------- ---------- ------------- ---------- —— - —-------- ------- ----- 0-291 ———■ \i V. 1 1 1 .9 8 .5 6 2 .1 2 0 .3 8 9 ---------- ---------- ---------- — 1 1 .0 1 0 .9 1 1 .1 1 1 .5 1 1 .0 1 0 .5 1 1 .7 8 .6 2 4 .7 2 7-19 8 .8 1 6 .3 8 7-31 6 .9 8 2 .1 5 1-31 1-71 1 .8 6 1 .8 7 1 .5 4 1*53 — —— 0.387 0.371 .Appendix T able 20 B lood P ic t u r e o f Animal Ifumber A78— Grotqp 5 Whole Blond Red Blood Hemo­ C e ll Count g lo b in Age day s 292 306 320 348 350 357 364 371 378 385 392 398 405 413 419 433 440 447 454 46l Red Blood C e ll V ol. Mean Corpus­ c u l a r Volume M/mm^ gm ft 2 u3 8 .9 8 9 .0 5 9 .2 0 9 -35 8 .7 5 8 .0 9 7 .5 0 8 .4 5 7 .6 5 8 .0 0 8 .3 5 7 .9 7 7.60 8 .1 7 7 .9 7 8 .8 7 8 .3 7 9-77 9 .7 0 10.50 1 2 .83 1 2 .3 5 12.20 12.77 10.43 10.13 9»40 9-70 9.33 1 0 .13 9-83 11.00 11.00 10.20 11.50 12.00 11-75 24.0 24.0 2 6 .0 3 4 .0 29-5 2 9 .5 3 1 .0 24.0 24.0 22.0 23*0 23.0 25*5 2 5-5 28.0 29.0 29.0 2 9 .5 29.0 2 8 .5 ----------3 7.9 3 2 .6 3 2 .1 3 3 .2 2 7 .4 2 9 .7 29.3 27-2 3 0 .1 31-9 3 0 .5 35-1 3 8 .2 3 5 .5 37.0 3 2 .7 3 4 .1 Mean Corpus­ c u la r Hemoglob in ung ---------14 .3 13-6 13-3 13-7 1 1 .9 1 2 .5 1 2 .5 1 1 .5 1 2 .2 1 2.7 1 1.8 13-8 1 4 .5 1 2 .5 1 4 .4 13 -5 1 4.0 Mean C orousc u la r Hemo­ g lo b in Cone. * 4 0 .7 4 0 .4 4 0 .4 3 7 .7 4 1 .9 4 1 .4 4 1 .2 4 3 .5 4 2 .2 4 2 .7 4 2 .2 4 0 .6 39-7 3 8 .5 39-3 37 .9 3 5 .2 39.0 4 1 .4 4 1 .2 Cal­ cium P lasm a In o rg . P h o s- Magnep h o ru s sium A sc o rb ic Acid mg ft mg ft mg ft mg ft 1 0 .3 1 1 .1 1 1 .5 12-7 ----------1 2 .0 --------------------12-0 -----1 0 .8 12.0 1 0 .2 1 1 .0 1 0 .7 1 1 .5 1 1 .1 7 .3 5 4 .6 8 4 .9 2 5-71 ------— 6 .6 5 ----------—------- 6 .7 9 1*59 1 .8 7 1 .7 9 2 .5 5 ----------1 .6 2 - —_ ----------_— 2 . 6o — — 2 .1 5 3.10 1 .7 7 1 .67 ?-55 2 .62 2 .0 7 ______ 0.521 0 .5 5 6 0.5 9 1 --------------0 .5 4 4 ------------- -------- 6 .4 8 5-00 6 .5 8 7 .23 6 .28 6 .6 5 6.83 0.4 9 0 -------—,------- ------- — 0 .4 3 8 0 .4 4 6 vXJ ■p- .Appendix T able 21 Blood P ic tu r e o f Animal Humber A80— Group 5 Wh o le Blood___________________ Red Blood HemoBed Blood C e ll Count g lo b in C e ll V ol. Age day 8 228 242 256 284 286 293 300 307 314 321 328 334 341 349 355 369 376 383 390 397 M/mm3 __i _ ---_ 7 .5 2 8 .7 5 9 .9 0 10.80 8 .8 9 7-80 9 .5 5 8 .3 0 8 .0 9 9-63 10.17 9 .9 9 8 .7 8 6.81* 8 .1 6 7.60 7 .6 0 gm £ 1 2 .1*5 10.87 10.1*3 11 .30 11.30 10.80 12.70 1 0 .2 7 11.50 10 .8 7 1 0 .87 10.50 1 0.93 1 1 .0 7 12.27 1 0 .57 9 .9 0 11.50 11.60 11.60 Mean C orpus1r u le r Volume i 3 1 .0 2 7 .5 2 1 .5 29.0 28.0 2 9 .5 31**0 25.O 27.0 21*. 5 26.0 21*.0 21*. 5 2 8 .5 3 0 .0 29-0 27.0 28.0 2 6 .5 2 6-5 Mean C orpusc u l a r Hemo­ g lo b in uug ----- — ------- —--------- — 3 8 .6 3 2 .0 29.8 3 1 .5 28.1 3l*.6 2 5.7 31-3 2 9 .7 2 5 . 1* 28.0 30.0 33-0 3 9 .5 3l*.3 31*-9 34-9 1 5.0 1 2 .9 1 0 .9 1 1 .8 1 1 .6 li*-7 ll .l * 13-1 13.0 11.3 1 0 .9 1 2 .3 12.0 li*. 5 l! * .l 15-3 15-3 Mean C orpusc u la r Hemo­ g lo b in Cone • i 1*0.2 39-5 4 8 .5 3 9 .0 1*0 . 1* 3 6 .6 37.1* 1*1.1 1*2.6 1*4.1* 4 1 .8 1*3-8 4 4 .6 3 3 .8 1*0-9 3 6 .4 3 6.7 4 1 .1 4 3 .8 4 3 .8 C alcium Plasm a in o r g . P h o s- Magnesium p h o ru s A sco rb ic A cid mg $ mg $ mg £ 1 2 .0 1 1 .4 11 -5 1 2.3 7 .4 9 6 .91 7-10 8 .28 2.13 2 .0 6 1-9 5 2.5 5 ------------- ------------- ------------- ----------------- 1 1 .9 7-96 2 .4 2 0.2 4 1 ------------------------- -------------— ------- --------------------------- --------------------------------- ®S ; ___ 0 .5 5 2 0 .7 3 3 0 .5 1 2 1 1 .5 7-19 2 .1 5 0 .4 2 8 ------- -—— ------- 11 .7 1 1 .7 9 .1 12 .1 1 1 .7 1 0 .5 1 0 .7 8 .3 9 7 .8 6 8 .2 3 7-31 5-93 7 .3 1 7 .9 1 3.20 2.19 1.97 2 .2 5 1.6 0 2 .1 2 2.00 ------------------------ —— — 0.533 0 .8 1 4 Apnendix T able 22 Blood P i c tu r e n f Animal Number A72— Group 6 Whole Blood Bed Blood Hemo­ C e ll Count g lo b in Age d ay s 395 409 423 451 453 460 467 474 481 488 495 501 508 516 522 536 543 550 557 564 M/mm3 — ----------------------- 1 0 .5 5 10.90 1 0 .0 2 1 2 .8 5 9 .0 5 0 .8 5 9 .5 0 12.30 12.30 10.70 10-75 9 .9 3 1 0 .2 4 1 1 .0 2 11 0 .1 5 1 0 . 1* gro j> 1 2.00 11.60 1 1 .7 5 1 2 .8 3 12.90 1 2 .00 1 2 .77 1 1 .50 1 3 .2 5 11 .67 1 2 .3 5 1 4 .4 5 15.10 1 7 .2 5 13-70 13-77 1 2 .83 1 3 .1 7 1 4.80 14.80 Bed Blood C e ll V ol. Kean- Corpus­ c u la r Volume $ 29.0 29.0 2 7 .5 3 2 .0 Mean Corpus­ c u la r Hemo­ g lo b in uug . . . . ------------------- ---------- -------- 30.3 -------------- — 2 9 .5 3 0 .0 27.0 3 1 -c 27-5 29.0 3 6 .5 3 6 .0 4 2 .0 3 8 .0 35-0 3 4 .0 3 2 .0 3 5 .5 37.0 2 9 .4 2 3.3 29.8 1 2 .2 1 1 .8 1 2 .0 9 -9 1 2 .7 _ _ _ _ — ---------- 3 1 .1 3 0 .5 29 .7 2 9 .3 39 .3 350 3 5 .2 33-2 29-0 3 5 .0 3 6 .5 13-2 13-0 1 1 .7 1 2 .3 1 6 .1 1 2 .7 13-9 1 2 .5 1 2 .0 1 4 .6 1 4 .6 . Mean Corpus­ c u la r Hemo­ g lo b in Cone. * Cal­ cium mg % Plasm a In o r g . Phos­ p h o ru s mg £ Mag­ ne­ sium mg $ A sco rb ic Acid mg mg f> 1 1 .4 1 1.3 13. c 1 2 .3 8 .6 8 6 .10 6.6 9 7-4 4 1 .8 6 1 .6 4 1 .9 2 2 .13 _____ ----- — ------- —----------- 1 1 .6 -----— ----------- — 0 .3 3 6 0.517 0.4 9 6 ------- ----------------- 7 .3 5 2.20 0 .4 5 5 ----- — —--- — ------—----- ------------------------- ------ — —------—--------- --- — 1 1 .4 6 .1 9 1 .9 0 0 .4 6 7 —----- ------- —--- — --------- 1 2 .7 1 0 .5 1 1 .7 1 2 .1 1 1 .8 1 1 .9 11-3 5-74 6 .4 4 6.79 7 .7 1 5-71 6.41 8 .0 1 1 .8 1 1 .2 6 1 .7 7 1.80 1 .76 1 .9 1 1 .7 4 ---------- -------- 0 .3 8 7 0 .5 3 6 -o "O Apnendix T able 24 Blood P i c t u r e o f Animal Humber A77— Group 6 Whole Blood____________________ Red Blood HemoRed Blood Mean CorpusC e ll Count g lo b in Cie l l V ol. c u la r Volume Age days 294 308 322 350 352 359 366 373 380 387 394 400 407 415 421 435 442 449 456 463 M/ mm-3 ----------------------- ---- 7 .8 1 — ---------- 8 .7 5 8 .7 5 9-07 10.70 8 .5 5 8 .2 1 7 .2 5 7 .5 0 8 .0 2 8.18 7 .2 7 8 .4 4 8 .2 8 -------------- 9 .1 8 gm $> 1 0 .2 7 9-70 10.50 11.60 1 1 .67 12 .23 11.60 10-93 1 1 .50 10-20 9 .3 3 9-47 i c . 13 11.43 11.50 10.87 1 1 .23 1 1.37 13.17 13.43 * 25.0 2 5 .5 25.0 3 0 .5 2 9 .5 29.0 29.0 27-0 27.0 2 5 .O 24.0 24.0 25.0 3 0 -5 29-0 28.0 3 2 .0 29.0 3 2-5 3 4 .0 u3 Mean C orpusc u la r Hemo­ g lo b in uug mm— — — -----------— — ----------- — — ------- 3 9 .1 1 4 .9 — —— — ----------- 3 3 .1 3 3 .1 29.8 25 .2 2 9 .2 29-2 33-1 33.3 38 .0 3 5 -5 3 8 -5 37-9 3 5 .0 1 4.0 1 3 .3 1 2 .1 1 0 .7 1 1 .9 1 1 .4 13-1 13-5 14.3 1 4 .1 15*0 13-3 1 3 .7 — - ------- -------------- 3 7 .0 1 4 .6 Plasm a__________________ In o rg . P h o s- Magne- A sco rb ic slum Acid p h o ru s Mean C orpusc u la r Hemo­ g lo b in Cone • C alcium i mg f> mg % mg j> mg i 4 1 .1 3 8 .0 4 2 .0 3 8.0 3 9 .6 4 2 .2 4 0 .0 4 0 .5 4 2 .6 4 0 .8 3 8 .9 39-5 40 -5 3 7 .5 3 9 .7 3 8 .8 3 5 .1 3 9 .2 4 0-5 3 9 .5 1 1 .0 1 2 .2 1 2 .5 1 1 .2 8 .1 2 6 .9 5 6 .4 4 7 .4 4 1 .8 7 1 .7 3 1 .9 9 2.43 0.433 0 .6 3 4 0-591 -------------- — ---------- --- -------------- --- -------------- -------------- 1 2 .0 7 .9 1 2 .3 6 0 .2 1 5 -------------- -------------- -------------- ---------- — . -------------- -------------- ---------- --- ------------------ -------------- -------------- — ---------- ------------------ -------------- -------------- -------------- -----------— — . ——— ------------------ 1 1 .8 5-34 1 .8 8 0-^93 -------------- -------------- -------------- ------------------ 1 1.8 1 2 .1 1 0 .2 1 1 .1 1 1 .1 10-9 1 1 .5 6 .7 9 5-17 6 .5 8 7 .7 1 6 .5 8 6-04 8 .4 5 1 .9 6 1.18 1.7 7 1 .8 3 2.20 1 .9 9 2 .1 6 ------------------ ---------- ------------------------ 0.463 0 .5 4 4 Apoendix T able 25 Blond P ic tu r e o f Animal Number A79—Group 6 Whole Blood_________ Bed Blood HemoBed Blood C e ll Count g lo b in C e ll V ol. Age days 285 299 313 3*4-1 3*6 350 357 3 64 371 378 385 391 398 4o6 412 426 433 4*40 *{47 454 M' mm3 gm i $ 9-21 7 .7 5 9 -35 1 1 .3 5 10.90 9-30 8 .4 5 7 .*40 7 .2 4 9 .^ 0 8 .6 5 7 .8 0 6 .1 5 7-59 8 .1 0 7-80 10-50 9 .3 3 10.50 11-30 1 0 .93 11-93 12.70 1 3 .2 5 1 3 .0 5 10.80 10 .87 9-70 1 0 . e7 1 2 .4 5 10.80 10.50 10.80 11.30 12.20 2 6 .5 25.0 2 6 .5 3 0 .5 2 6 .5 28.0 3 3 -5 3 3 .0 3*4'.0 28.0 2 5 .5 2 4 .5 2 7 .5 2 8 .5 28.0 29-0 3 1.0 2 7 .5 3 1 .0 ------- --------- ------- .------—--- — A Mean C orpusc u la r Volume u3 Mean C orpusc u la r Hemo­ g lo b in uug Mean C orousc u la r Hemo­ g lo b in Cone. 2 -----------— —----—--- - 33-1 3 4 .2 29.9 2 9 .5 3 0 .3 36 .6 33-1 3 4 .5 3 3 .8 2 8 .6 3 2 .9 35-9 35-6 4 0 .8 3 4.0 39-7 1 2 .3 1 4 .1 1 2 .8 1 1 .2 1 2 .2 1 4 .0 1 2 .8 1 4 .7 1 3-4 1 1 .3 1 4 .4 13-8 1 2 .9 1 4 .2 14.0 1 5.6 3 9 .6 39.3 3 9 .6 37.0 4 1 .2 4 2 .6 3 7 .9 4 0 .2 3 8 .4 3 8 .6 4 2 .6 39-6 39 -5 4 3 .7 3 8.6 3 6 .2 3 4 .8 4 1 .1 3 9 .4 ------- ------- ------- Celcium Plasm a________ In o rg . P h o s- Megne- Ascorbic A cid sium p h o ru s mg $ mg $ mg 5S mg $6 1 1 .0 ii* 5 11*5 1 2 .0 7 .0 6 6.51 8 .1 2 7 .4 4 2 .0 6 1 .9 2 1 .7 6 3-27 O.344 0 .5 2 5 0 .5 4 4 ------------- ------------- ------------- ----------------- 1 0 .9 5-79 1.76 0.4 3 9 ------------------------- ------------------------- ------------------------- --------------------------------- 1 1 .4 6 .58 1 .8 8 0 .551 ------- ------- ------- 1 0 .8 1 0 .7 9-1 1 0 .9 1 1 .2 1 0 .8 11-0 6 .3 2 5-17 7 . *;4 6 .1 0 5.90 5-74 7-71 2-04 1 .9 ? 1 .9 5 1 .8 5 2 .2 5 1 .9 6 2.0 2 ------------------------- --------- 0-387 0.413 - 100 - Appendix T able 26 Growth Record by A nim als ny» it. 186 193 203 195 183 188 196 200 208 227 222 207 234 253 238 262 268 266 281 307 302 309 332 3*+5 345 373 377 392 389 423 434 443 471 480 470 460 468 504 500 517 513 518 515 528 550 587 C681 Age doys Wt. lb . 229 180 250 187 194 237 241 201 208 256 245 215 222 249 229 251 236 253 243 257 252 250 248 256 264 • 262 272 274 283 278 286 287. 274 293 292 299 292 307 317 327 324 341 331 357 338 362 356 3*+5 370 352 392 359 366 *K)7 421 373 424 380 447 387 450 39*+ 470 401 481 4o8 415 477 498 422 4?8 429 494 436 443 535 450 545 457 565 464 561 471 530 4 ?8 567 485 582 492 598 499 634 C684 Age day 8 Wtlb . 166 173 180 187 194 201 208 215 222 229 236 242 250 258 264 272 279 285 293 303 310 317 324 331 338 3*+5 352 359 366 373 380 387 39*+ 401 b08 415 422 429 436 443 387 383 377 392 407 379 396 394 423 425 397 417 415 435 429 442 444 *+33 465 473 463 489 472 497 500 516 531 55 0 5*+9 572 589 605 606 634 628 660 642 675 667 713 C711 Age days Wt. lb . 84 91 98 105 112 119 126 133 140 147 15*+ 161 168 175 182 189 196 203 210 187 182 194 190 199 216 223 247 258 285 289 325 330 358 356 378 383 *K)5 410 C714 Age days 49 56 63 70 77 84 91 98 105 112 119 126 133 lb o 147 15*+ 161 168 175 184 189 196 203 210 217 224 - 101 - Appendix T ab le 26 - C ontinued Grov/th R ecord by Animals w t. lb . 78 85 93 98 101 109 114 118 124 136 133 151 147 163 172 170 182 194 205 216 224 241 C718 C719 C724 Age Wt._______ Age Wt._______Age d ay s l b . days l b . day s 49 56 63 70 77 84 91 98 105 112 119 126 133 143 147 154 163 168 175 182 189 196 203 210 217 164 168 185 195 214 225 242 249 266 269 304 314 303 330 362 379 415 408 455 465 46o 486 506 518 552 21 23 35 42 49 56 63 70 77 84 91 98 105 112 119 126 133 ik o 147 156 161 168 175 182 189 113 105 126 132 142 152 160 170 180 203 214 228 243 259 275 280 295 315 329 360 365 382 396 419 42 6 35 42 49 56 63 70 77 84 91 100 105 112 119 126 133 140 14? 154 160 167 174 181 188 195 202 209 tVt~T lb . 89 96 108 115 128 136 147 163 172 195 196 221 226 2k0 262 27 2 277 288 298 306 320 339 363 370 389 *K)4 i - 102 - Apnendix T able 26 - C ontinued c z ii bg>e _ daya 35 42 49 56 63 70 77 84 91 100 105 112 119 126 133 140 147 154 l6 l 167 174 181 188 195 202 209 217 224 230 237 244 251 258 265 272 279 286 293 301 308 315 322 329 336 343 350 Wt. lb . 133 145 149 167 174 183 197 212 224 249 250 260 266 2 70 282 289 290 305 311 322 342 349 365 373 379 402 410 413 4 l4 420 435 446 450 472 480 492 500 510 500 530 539 525 544 561 581 598 C726 Age days 35 42 49 56 63 70 79 84 91 98 105 112 119 126 133 140 146 153 160 167 174 181 188 196 203 209 C730 w t. lb . 82 77 76 80 85 94 103 110 111 119 121 137 140 150 162 166 185 196 212 222 239 248 265 257 262 266 223 230 237 244 251 258 267 272 280 287 294 301 308 315 322 285 292 292 307 326 335 319 320 340 355 356 390 383 400 413 428 329 436 216 0739 Age days w t. lb . Age days wt • lb . 40 47 54 6l 126 37 44 51 58 65 72 79 120 128 68 75 82 89 98 103 110 117 124 131 138 145 152 158 165 172 179 186 193 200 207 139 140 143 149 161 176 187 212 213 228 238 250 .272 275 281 302 312 320 345 353 353 373 389 397 86 94 101 107 114 121 128 135 142 149 156 163 170 178 185 192 199 206 213 220 227 152 160 160 173 188 198 202 213 227 230 240 255 263 272 285 295 300 313 311 335 339 351 351 356 374 388 - 103 Appendix T able 26 - C ontinued C74l Age d ay s C742 Wt ■______ Age lb . d ay s 0744 Wt._______Age lb . daya 35 42 49 56 63 70 78 85 91 98 105 112 119 126 133 140 147 154 162 169 176 183 190 197 204 211 132 148 160 166 178 182 193 182 203 215 234 249 262 2 67 287 286 303 319 324 342 341 360 362 372 386 413 134 143 158 170 179 191 203 218 235 240 270 281 300 315 335 340 356 388 *»01 393 425 431 467 498 511 527 35 42 49 56 63 70 78 85 91 98 105 112 119 126 133 l4 o 147 154 162 169 176 183 190 197 204 211 14 21 28 35 42 49 57 64 70 77 84 91 98 105 112 119 126 133 141 148 155 162 169 176 183 190 C749 Wt._______Age lb . day s 95 105 118 124 130 142 144 149 157 159 172 172 187 188 200 197 207 207 215 223 241 250 240 253 262 288 77 84 91 99 106 112 119 126 133 140 147 154 l6 l 168 175 183 190 197 204 211 218 22 5 232 W t. lb . 205 220 233 244 254 275 282 306 312 329 347 360 373 389 410 424 4o6 441 443 475 500 511 523 1 - 104 Appendix T able 26 - C ontinued jnrjLi— ib . A75 A73__________ Age Wt.______Age Wt. day 8 lb . day s l b . 547 515 485 551 559 565 575 582 593 634 670 656 694 697 685 691 720 73 6 716 722 695 731 730 721 733 375 382 389 396 403 410 417 424 431 438 445 452 459 466 473 480 487 494 501 508 515 522 529 536 543 433 442 453 440 430 449 473 463 496 532 560 565 572 593 592 597 621 620 681 603 615 609 623 623 637 327 334 341 348 355 362 369 376 383 390 397 404 411 418 425 432 439 446 453 46o 467 474 481 488 495 423 434 448 417 443 431 458 465 466 507 531 530 536 565 550 580 601 581 623 595 655 643 636 658 693 A76 Age days Wt. lb . 307 314 321 328 335 342 3^9 356 363 370 377 384 391 398 405 412 419 426 433 440 447 454 461 468 475 471 451 471 437 471 465 472 500 499 532 543 529 572 578 560 596 621 637 653 612 626 630 649 661 681 A - 105 Appendix T able 26 - C ontinued w t. lb . *4-39 1+32 1+60 422 1+57 1+51 1+66 1+65 1+96 503 51+O 51+7 551 556 600 571 597 612 627 580 591+ 577 592 635 680 "A78 ' Age d ey s A79 Wt._____ Age lb . deys 296 303 310 317 321+ 331 338 3l+5 352 359 1+57 1+56 1+70 1+32 366 373 380 387 391+ 1+01 1+08 1+15 1+22 1+29 1+36 1+1+3 1+50 1+57 1+61+ 1+60 1+51+ 1+91 1+98 509 51+5 569 561+ 601+ 597 602 619 61+5 639 636 631 623 637 660 61+5 662 289 296 303 310 317 321+ 331 338 31+5 352 359 366 373 380 387 391+ i+Ol bO8 1+15 1+22 1+29 1+36 1+1+3 1+50 1+57 A80 Wt.______ Age lb . deys 360 358 371+ 352 363 389 396 391+ 1+07 1+1+2 1+1+0 1+51+ 1+67 1+81 500 505 51+6 539 533 526 575 556 56 7 571+ 60 9 232 239 21+6 253 260 267 274 281 288 295 302 309 316 323 330 337 31+1+ 351 358 365 372 379 386 393 1+00 Wt. lb . 1+09 1+28 1+1+5 i+30 1+55 1+89 508 1+97 525 555 580 580 589 600 615 621+ 651+ 621 672 681 655 652 656 691 711 i - 10 6 - Appendix Table 27 Comparison o f Body W eight w ith th e R agsdale S tan d ard - Groups 1, 2, 3* ana 4 . Weight We ig h t Age A c tu a l S td . Age A c tu a l S td . lb . lb . lb . lb . d eys days Group 1 ---1+9 164 ---552 217 Animal Number B re ed C718 B r . S«- M C719 A yr. M 21 113 95 189 426 338 ------ C724 Je r. M 35 89 82 209 404 325 ------ C?42 H o i. M 35 134 132 2.11 527 458 ------ C749 G uern• M 77 205 132 232 5# 386 ------ C715 Je r. M 35 42 78 85 85 241 88 285 -----Co C739 H o i. M 37 53 120 154 134 155 53 227 154 388 155 489 Co Sex Group 2 42 82 88 182 Supplement ------ C?4l H o i. M 35 133 132 237 132 298 133 211 287 413 298 458 Co Co & Pe C744 H oi. M 14 29 95 119 108 124 29 190 119 288 124 418 Co 84 88 158 18? 184 310 Group 3 88 196 211 158 210 353 184 310 410 211 353 456 Co & S u lfa Co 35 50 165 133 152 317 132 151 368 50 165 350 152 317 598 151 368 722 Co & S u lfa Co 40 47 6l 163 126 139 143 318 138 147 166 363 47 61 163 207 139 143 318 397 147 166 363 450 Co Co & S u lfa Co 49 84 123 155 206 300 Group 4 84 150 204 123 224 276 206 300 470 204 276 483 S u lfa Co 35 57 132 82 81 ?50 118 144 359 81 250 436 144 359 592 0711 H o i. 07 25 H oi. C730 H oi. C714 0726 H oi. H o i. M M M K P 57 182 329 lll , S u lfa Co i - 107 - Appendix T able 28 Com parison o f Body W eight v i t h t h e R a g s d a le S ta n d a r d — Groups 5 ond Animal Number B reed A73 H o i. F 375 1*31 Groun 5 433 652 1*96 720 1*31 5*+3 1*96 637 720 81*8 A75 H o i. F 327 333 423 1*66 539 662 383 1*95 1*66 693 662 796 Sex Age d ays Weight A ctu al S td . lb . lb . Age d ays Weight A ctu al S td . lb . lb . Supplement ------- ____ A78 H o i. F 29 6 352 *+57 509 51*6 622 352 1*61* 509 662 622 763 ------- A80 H o i. F 232 288 1*09 525 1*1*8 535 288 1*00 525 711 535 682 ------- A72 H o i. F 399 ^55 Groun 6 51*7 681 593 752 1*55 56 7 593 733 752 875 Co 562 636 363 1*75 1*99 681 636 776 Co A76 A77 A79 H o i. H o i. H o i. F F F 1*71 6 307 363 4 99 298 35*+ 1*39 1*96 5**9 621* 35*+ 1*66 1+96 680 621* 765 Co 289 3**5 360 1*07 536 613 31*5 1*57 1*07 609 613 75*+ Co