INFLUENCE ON THE GROWTH OF RATS OF VARIOUS MODIFICATIONS OF THE SHERMAN-CHASE DIET FOR A VITAMIN B1 ASSAY Thesis for the Degree of M. 5.. MICHIGAN STATE. COLLEGE Marian R. Weaver 1940 h \ I at.“ l \ .0"' 0 0A .11; '1. ' .fiwmow. I 3" 'J p .-I I, l" -_.~' 7'..‘A V I . Offlg} \. ‘ , ".cl- ' r‘>I . w a x i 469‘}??? »; * L, M ‘ . . ' f . .47 J u/ _ ’ -- ‘.l‘lr .. i“ ' '1 —.' I s l ‘x .' I ‘J ' ‘5 _. f \ .. a?" s. ‘1. .* ..;- . . - .A'. . ‘1' _.'.' ‘ "$Il; ”‘3'" “_- . . ‘.\' a. I .z‘J‘c' , -. “.4 "(.‘$ V, . " ‘<-‘ ’ ‘ ' T I .U’ ‘ ' l .' “:‘. '1, ‘2' - _ ‘ . ‘ -_‘ ‘ R“ i. J “.5213"; Lug-w o .‘_ a o . ‘ ~ ‘ “4 I. ‘ . . “ k "v". . I. "I . ‘-.-‘ . (4.. c3 1‘.’ ." . 2.“ , a . “- . -. 1"; "’v‘ '0..er ‘ ». , ~ 3 -’ “I ' .‘~ ' -.r:{- "f: '- ‘l . I . {:K.‘. >‘:‘_"""‘~.6“ “.1. - ‘ A‘t. -'-.MV u,~.‘.' ,‘ 4 r I l I i L; i _a I :J'-’Li y‘d-"K.)lu u, ’. r‘ a ‘; ":‘ér. ‘4. . . .34: ‘ .... _ . 3'11. .1 £_ '7 E'W ' 1'" . E .' f" \ K 5 "‘1 .pl' '. :3 ._ r‘ ‘. ;' "r 31%. 1»- r, W. “.A., I.“.‘ .. 2-. " w¢ "w 'I -. '0 c-yw. luau - . ‘rl ‘13”). ' “’37 IIIELUEIT E on THE GHQ-v.2: c: RATS CF mmous MODIFICATIOIT- ’ OF '33 SHERMAE—CHIQE DIET FOR A VIT KIN B ASSAY l E by marian R. Weaver A THESIS Subm'tted to the Graduate School of Michigan State College of Agriculture and Applied Sclence in partial fulfilment of the requirements for the degree of EASTER OF SCIEHCE Department of Foods and Nutrition Division f Home Economics 191m TH 5:513 - "Tfi'f‘W‘T r m -;‘7$T A‘czuxmgzp 7.--.‘_.-. The author is much indebted to Kiss Flora H. Hann'ng of the Home Economics Division of Kichigen State College, under whose sugervision this studv was carried on, for her mini criticisms and helpful suggestions, and to Dr, Earle Dye, Dean of the Division of Home Economics of nicnigan itate Colle;e for her heloful suggestions. Acknowledgment is else given to Wilson and Comoany, Chicago, Illinois, for generously donating the Factor W '5 ’1 (D 'd m H “J a H. O {3 0 129323 “A F“ n-wfir'er “at ,1! . ( -iLJHA—l b--. ‘dJIA- (:4. 4 D Introduction .......................... 3 \ fi_ 0 .' a ’L. .- ~ , I.L.'Vl‘3.‘.' O: 1.91" tulre O o o o o o o o o a o o o o o a o 0 LI- fl-"'~ .W A‘ -_- | 7‘. :1..- ,C‘Tl..lel.‘aC.1 ’JTCCCJLT o u o o o o o o a o o o o o o o 13 ~~ 4. ,— . .1 2 ' ‘ .. ' ,. (7 RCQ’JILS (”IF-.A. CLISC‘JSQIJn cocoon-cocoooooo l'J - n n7 SLIITTI’L‘I'}; . o o o o o o o o o o o o o o o o o a o o o o o o o O o o g o L. T.‘ '1. ' . .. —‘.~. "3' .. l ‘IIOJTJ: ”:17 o o o o o o O o o o o o o o o o o o o o o o o o o a _) ALTJEDenI-llx 0000000000.000000000000000-000 33 IL “L nTCE ON THE GROWTH CF RATS OF ViilCUS MODIFICATiCTS 5w OF THE S":ELAn-C" SE DIE T FOR A VITAIIH B ASSAY The Sherman-Che: e (l) diet for assaying food for vitamin B1 (thiamin) by the rat growth method has been used since 1931, \vhen the sntineuritic vitamin and a gr “wti factor contained in 335 st vere tr ought ohe the only vitamin B factors necessary for rat growth, more recent— ly, studi on several other vi :..in B comgoaents have been puoli she', q Some of these fa ctors may be low or lacking in the besel diet, which \J ends on autoclaved yerst for the B factors other than thi.' min. With the evailability of ma ny of th 1688 vitamin 3 factors in crystalline form 't egpeered cossible to test wlietter the antoclered yeast in the Sher- rn-Chese diet was supplying. all these B factors in amounts necesser for normal growth. Therefore this study was undertsken to determine which of the vitamin B factors, in addition to the basal diet contain- ing autoclaved ye net and thiemin chloride,wonld in‘nce better growth under our laboratory conditions, and if sn nlfi te tree ted greest replac- inc tne sutoclaved yeast would give a more conglete basal diet The use of the growth of standardized young rets as a 29:28 of detecting the :rese cc of vi anin 31 in foes“ Wes early erggested, and a method w-s suggested by Sherman and Spohn (2) in l 23 The basal diet develOPed ‘3 them was gen er “11? ~ ed Up to the time when it wrs demonstrated that for norral growth the rat recnired in as oi tio n to vitrnin £1 a heat stable fraction remaining in autoclaved 3e: st. The Sherman-Spohn diet was then modified b*r vhcse (Sherman-Chase diet) (1) that vitarin 31 was considered then to be the first if not the only growth limiting factor. The Use of the preliniizry depletion period "as included as a,;rec anti onarv xea sure, During the 1; st few veg s, it has been definitely established that the heat stable fraction of yeast (calledB , is a mixture of a nerer of vita: ins. niboflavin, nicotinic acid, vitamin D; (pyridox- J ne), Factor W, pantothenic acid, "filtrate factor" are snhstances ’40 I”). :3 (1' SJ (3 r4. '1 H (T) l.) 1* O :3 d' O 4 H cf' ”a to 0 (/1 9 6 C ('9' ’3‘, (D ’1 (‘D which have been studied “3 factcrs a re essential or growth of the rat, others me V or ma V not be essential. The question has been raised whether or not all these frac- tions are sugplied in autoclnved yeast in amounts necc ssarv for ooti- mum growth, and whether differences in laboratory manipulation would influence the anounts in the au ocleved 3east. Following the denonstration bv Goldberger and Lillie (3) and by S:nith and ITer.5r ich (M) of tne indelendent existence of what is now reestnized es the conglex B apb "t from the entinenritic vitamin 31, the bggothes s was widely acceated th at vita :nin 32 wc“ d pre event or \J’] ~ o o ‘ ‘ ‘ , .. r v .~ ~ -.‘ n v 9 Cure rat dermatitis, Cerine blacn tongue, Lnd hun.n Lellepru, ”hng were consequently re5a rded as strictly ene105ous diseases. In 1330, LI) 0 (5) produced exgerinintnl evidence to the efi._ B was a connlex, ocmi>osed of two dietarv essentials-«vitamin B and o “ *1" ‘ f ' nresent in anteclaved veest. Sure, Smith and nlk's (0) experimental work also Showed ty~' Gyor5y Kuhn, end Wegner~Jaure55 (7) showed that riboflavin (lcctofl'vin .N I‘ r:- ': L ‘ “ '3 1‘ ‘: I" ‘: '. . T A y . M ; nd ovoflavin) nosse:,set -ntonse titumin nativitj. Very soon c.9r5d (5) A. (T) dCJOD? treted that rihof CV; was essential for rat growth but distinct from the :it cer1a titis greventive fact or (BG)' Cnicx, oogjin5 and Ed5e ‘ (3) confirmed Gvor*"'s woik, and Le ficovsky and dukes (10) also proved it necessary fci growth in rats and chicks. nitely known that riboflavin is necess: y for normal growth of the rat. Hcflcnry and G avin (11), r Birch (12) used 13 *a.4u of riboflavin in iet, th0t5h they did not include autoclaved ve not, G {2. th ir vitamin B free for satisfactory 5rowth. .J L) When Goldberger and Lillie (3), workin5 on umen pe11e5ra, found a a factor ('-P f'ctor) present in yeast which Lred the disease, they 0 thou5" ht rs t dermatitis wes caused by the lack of the same factor, tlou5h Ht 53 since lWeen proven b;» Gyorg (8 ) h never thst rat dermatitis is vitamin B . So it was not until Elvehjen, laddedn ed nicotinic acid and found that it was U) d H O ,3 0 "3 L.- b O H .-.J (D a, A H V F“ m 0 1—4 $1 (1' snecific for black tonsue in dors, anelesous to human nellecrd that .. -— w 9 exact difference between the tn 0 tzrpes of dermatites was recognized. C (D Oleflwn Bird, Elvehjen and Hart (In) th ou5ht nicotinic acid “i5ht be 0-,: , essential for growth of rats. Howevs r Damn and Suhbarow (15), also oirch us) ad-sd 1 milligram of nicotinic acid to the B1 test diet and cid ('1‘ found no increase in gain in weight, thus concluded that nicotinic was not essential for growth of the rot. Dimick exgresses the theory "the rndonbtedly nicotinic acid must 313* a role in rat metabolis a‘ cince it form a y-rt of the enzvue c"rs—"ten involving hydrogen trsnszer in the metabolism of carbohyarates. Either, like vitamin C it can be synthesized by the rat or its stores are so well consc rved as to render e:’h* stion from the rat body tissues difficult or imzoss ible." - ..-(_ As indice ted shove vitaznin 3 wrevents and cores 5 nutritior Ll fiernititis in rats (Gyorgy (3)), ard Leokcvsky (13 ) sho ; ed that the addition of it to the list caused a res um; tion of growth. It is pro- CCpping (19) with vitamin H o: Ifo.x n (so) and of Booher (21), and with %ctor I crystallized. bv Leak vsky (22) * A recent naner by Dimmich aha Scheffler (2N) showed that rats receiving rystalling Factor I (BS) utilized food better tlsn those on ( the defic 1t diet. ine” ionnd that 73 .na of crystalling vitamin P6 satisfied the rot reruir enent for nc-nrl frOhuh n :35 Elvehj em, Koehn, sni Clcson (£5) srosented evidence of the existence in liver of a thermolahile substance necessary for the growth of rate Mi h they believed to be different from any memsor the B complex previously described, and they progosed the name Factor *In 1939 Gvorg v (23)a1d Eckherdt er“- es te d that in accordance with the che:nicel neture of vitamin 3.: the te r; .1 p;. rido: ine to be an agyprooriate name. This year (13M0)et the AHlC ricen Institute of Intri- tio n it was recommends d hv the Co:uittee on V1 t 21in Nomenclature that vit; min 36 be Ciu led "Pyrido: :ine. W for this substance. It is an alcohol-ether TTeClUI t; t9 from water extract of hog liver, and is n:et labile. Eline, Tolle, and Folsom (Co) believed that autoclaved yeast, as the sole source of vitanin E -- ‘7 ‘\ ‘.r~ FPA. ‘: ‘ "_ ‘ A. 1 'V r ‘ COfi 19* Ct??? tflufi Vitewln will oe lo. or devoid c: :ector a as it ' j . ' ‘fi ‘ I'~ ‘ . "~ \ m.” . v. '1 ov- ‘r‘ ‘N f‘ "‘ - «8d 1m tuG Hehblu TFCCcss. inese n rkers ceionstrdteo tgflt f liver extr: ct to a vitarin free diet nlus thiamin, C’- (D 9 Li {‘1‘ +4. ,1. b4 u ) :3 O synthetic rit01lsvin, carotene, nicotinic coin, choline increased the nos been made of the crvstollization of Factor X. g “e vi;r13ns nreeerstions frtn l1ver may I ‘ (‘9' ’1 c \ ’3 a 9 U) 0 ’1 (’1' Uillia*s, T"m‘n Goodyear, Trues"”ll and Holaday (27) announced the discover? of a groutn Coterninnnt of univer e1 tiolo c l occnrdnce vh1c1 'd~ n'neo "pantothenic acid " It is it~ 1f uni Able h t con oe concentreted from liver extract or rice bran extract. Williams (9?) sncnsihle for tart of the growth pr noting activity of liver extract or rots. Oleson, uoolleg.’ and Elvehjem (EC) found that rats needed ‘ Factor W as well as pantothenic acid for rat growth in ad d6. 1tion to the crystalline factors (thismin, riboflavin, vitamin 3/) which were sup- 1) snoxed thrt tie tn 0 filtrste foc- plied in the basal diet. Jukes tors--rentotien1c iCld b :,”3) et. al., and_ tqc ch icv d9?- 0 0 ~ ' P v ’ 0 1P1 w I ‘ 1 ~_ matit1s factor oy noolley, weisran, and ;lvenJem (34) are proonuly identical. Therefore there are irdicotions t 't not" p:ntothenic acid actor W are necessary for iormsl 3rowth t dough tie quantities "' of foods for vitamin 31, all these (/1 5 necessary Iactors should be included inadequate amounts In the oner- man—Chase die et, autoclaved yeast is the reco Coward, Tarn, Ling, and or3an (E ) found tnat with increased amounts of autoclaved yeast from 3 percent to 23 percent of the hasa diet tne resultin3 3rowth would be 3reater with the s. 0 amount of vitamin E H - a added. This 8.101ed tr e in terdene ncenoe of the vitamins end t1at t‘m o q o estiration of the amount of t‘nev1tanin 31 in a food ne1n3 tested *oald ) vary with the sun ly of the vitamin Bo factors. Some other metho‘ of L'- yeast preparation may he used and hc1s leen su33ested.1he lability of C'- hiamin in neutral medium is well exemo if ied by the ease of cleavage hv sulfite ct pL 5 to 6. (Williams, Z.atermrn, Kenztesv, Buchaan \36)) into two inactive suostanc ces. Kline, Tolle, and Frlson (77) used the above observations as the bases for treatin3 yeast. The1r dietnas toe same as that 13 ed bv Sierman-Chase except the yeastiflustreated with suifite. The animals grew as well as those receiving autoclaved yeast. 1 / ~1\ Booker and Hart; ler \ju/ found that they n vita— *3‘ {1 $1: cr- 0 (D p .3 CD "5 D ”’1 L .1 d H. c—r min 3 assavin3 o1 foods when the diet contained sulfite 'te' "l as when it contained sutoclaved yeast, and that the choice between sul— 1"? r21 (14 1, .1» to n c+ (‘1 ’- fite-trertted yeast and au- oclaved " ast could be decided on the Lesis of convenience of the exnerir en u. Similarily Smith and Cimmich (33) recommended the use of rice bran concentrates treated with sulfur dio- xide s an ition to the Sherman—Chase diet as it nrovided members {11 of the B complex which are often not supplied by the autoclaved ;ree st. {:2 a It is ev vi .en nt from the work reported thrt the Sherman—Chase diet with the addition of autoclaved yeast is probetlr low in Factor W and 1 I. ,antothenic acid and the addition of actor a nrenaration to the diet } .1 C) ‘erlrrr"l Procedure 3}; ; In this expcrinent, l4 litters of white rats of known herititv " " . n .7 ,‘L I ,. A... MI“. ”‘1', ‘r‘~ xere used as ex~er11‘1tu- enlezls. h¢b3 at tne “Q0 of g; to g- on}: .7 p . ' A , ‘lf- r— ‘ _ r 9 O o 5. 0 end I o yew at of “reroiln.telg 3 to E; ora~s c,re pl.ce« 1n 1n11v1- . 0 II 7" ’- O N M111 on 3 With raised screen oottons of 9/10 inch mesa, w clevned t ice weekly. The? mere fed the vitonin 7 deficient diet for ’) shout 2 weeks to defilete the bed? stores 1f the Vite min. water was 9110? ed ad lititum. After one week, harnesses (Page (h3)) known to interfere with a roliatle vitenin 31 assa et. al.). There is evidence thet vitamir 7 is formed if the cecum, probbhly as the re ult of oacte erinl action on the reed residue there. (D Guerrant and Dutcher (hf) found that the vitamin 31 due to refection is not absorbed from the lo er i1mtti11cl tr ct tut the rat may obtain :»er efit fro m it by the simple exp ient of conemi mi 1 his feces. The use of harnesses tends to prevent this. The consumrtion of the basal diet was controlled during the entire test so that the rats witoin each litter were allowed a uniform amount of the basal diet thus containin' the same amount of the B factors of S autoclaved yeast. The food mes weighed twice weekly and each rat re- ceived the amount of food for the next half week that the rat with the lov.est food consumntion }1ad eaten in the previous period. If all the rats in the litter had eaten all the food allowed it would be increased ‘ by 5 grams in the next period. This amount of food was o'vided over ‘ the half week consumption perio o. ll After the first week, the rats were weighed every other day and sometimes each day toward the end of the second week. The weight of the rats usually decreased after harnessing but would increase again my the fourth day. When tLe weight of all the rats in a litter went down the second time or remained stationery, that litter was cons1dered to have been depleted in vitamin B . lne depleted rats Were then placed on sufielenent, weighed weekly, and tne food consumntion recorded bi— 1arnesses were removed at the time of weighing to allow tie animals to clean themselves, and were not reeleced for several hours. When a litter of rats were dEUletEQ of their stores of vitamin s1, .11., x «1—. '6 vw‘ '31 fiv' . I. $.18 n.) 98]? O.L L “431'? Latl'l ndle rats and mean Helght in sec 1 group were distributed equally. The six groups were fed as folloxs: * - -- -- \ ....., n'r‘ - '* r - . ~ -‘ l. easel diet plus 4 Dane; tnidmin chloride (Cantrol Uroug, - q ‘3 R N “- v3 f L1 . ‘rv‘. .I‘ . "1 ‘\ A ‘. ‘ T. Lds l r-ct fills u é-‘:s .Llawln clloride plus 1 mgn. nicotlnic 4‘ r—J 7 -:r r1 . aw ' ‘fiv‘y‘1‘ L. .l‘ .* ..' . " , h vs. ~ . - ;. stal Llet plds r waxed talemil cnlorioe plus lo bemma riuo— P" ”vim. ‘d-Lv I -~ h. Basal diet glue e games tfiiemin chloride rlus lo gamma Vite- o ‘ O C O 1 I." I“ . a ' 4—.-. 'v‘" ‘ ‘- - “V P --~ “N ;. Edsel diet 91‘: e Den l t lo 1n caloriue plus 4.7 on. gtctcr r' , .- a eer nee”. f ~‘~~Q 3‘: ~ . ‘- Q. ‘ ‘K a. V 0" I~ .. .- p. ”allilte JCfiSb" Olet plus 4 Jenna tnltaln Chlorlde. Mn 0 1 n q ‘ ' o n "C V ~4_ lee subplements were continued Ior tn, 4 weeks test seriou. thl tne r"cepticn of the Factor W er.feration, the supvlements were fed daily '4 as 1 -7 12 includinr Sunday by a calibrated oi; J-ette directly into the rioutl of the strength and each day's p02tiin rue diluted with a 1? percent suQQr solution. The rut: ate vith avidity the 0.35 grams of Facter H pre- paration which wns Wei'h d on a sensitive torsion inlance, end fed twice a week in a s all container All sup:lelm'1ts with the exception of Factor B preper :.tion, were male us in stoca solutions and kept in tne re Hfri gerL‘ to and fron the stock solutions the exact dilutions were made up each week. ”‘11 ' "rt“ limo-n thd-aw’“ “1 w'dn v'r‘c "\ "1‘ P 3:3 :- 7‘ J.‘.'e SOl‘mtlofl O: cr‘y’u V. ‘oLQ ‘ALye All]. CIA-510‘] Q ul';+..) Lrt pwrt'hL Lis J-eC'DPP :10 mended by the Inte rrntioral Connittee on Vitamin S dadurd cat ion of th: J solution "as nre_eared l; dissolving 23 milligrams of thiamin clloride (U.S.P. vitamin El refzrcree s 3rd) in 10’ ml. of e 25 percent aqueous ethyl alcohol to hlcn mes "1 A ‘ V"' ‘ "‘ fi , "%V‘ Vw'v P p .E' . — H loued Hol to mane tne eolttion c Froninltely u/5OO ac1d. Then, 0 ml. :" F1 H (D ,0. Q7 C‘" (D "3 O of this stock solution were diluted to 50 ml. with die The rats were fed l/M ml. of the final solution plus l/M ml. of the sugar solution which equalled M games of thienin. The stock solution of crvst lline riboflavin (Merck and Co.) was nrenered by dissolvir; 23 milligrams of the riboflavin in lo~ ml. H/ECO £01. The solution was ke;t in a brown bottle in the refrigerator to prevent deconnosition by light. Each week 13 ml. of the stock solu- tion Has diluted to 50 n with d stilled water Thus l/M ml. of the final solution plus l/h ml. of tie suonr solution equalled 13 gamma riboflavin the daily dosage for the rats. 13 L. equalled l milligram doily. To make the stock solution of crystalline vitamin B, (ovrilozine) , 29 milli;rems were dissolved in 100 ml. of distilled water and kept in a dark bottle. Each week 23 ml. of the stock solu- tion was diluted to 50 ml. with distilled water. Thus, l/M ml. of tie pantothenic acid, and will sup ly the various memoers of the B couplex "1 H- (‘1’ ) J '_\ .1. a one exception of thianin chloride.* The animals received 0.7 1 is about equal to 3 percent of the Leocl O'et us recommendri l" Elvohjem.* T1 SLer.ou-Chaso "‘t"r1o J —~deficient diet used hod the follow— in; composition: i'.-.:.or~ ° 17” fitdnin y iree casein is. Salt mixture (Hank-Cser '31) 4: O \n Filtered butterfat (Jul 0 C) Cod liver oil 2.3 Autoclaved yeast Sulfited ytest 11.0 Cornstarch 53.0 (n (1 H. 9. *Private coununicotion with Professor C. A. Elvehjem and Dr. Di ." 1 The casein nos ireed of vi auin 31 by cola extraction nitn e perc ce. t etnfl alcohol (t3 wei or} . Qf fl ,s‘; ”<9, I: K“,\ '3 17-. o (C‘\ .1/“111 -- r. c A ~' r} .~, ~ 5‘ ' “rm . IL 5 .< t 3'1 Misd- D" b‘JJl.LL L.n L 9-1-1.1.;«fl L.’ ’ J‘s/U Org-~11”): OI C‘Jvrse CM-‘J‘vln ly‘LlS ULe(‘. [rt liters of 03 percent alcohol, and the stole was shsken for l , ‘v. 1 - ‘ ‘3‘ ,' '1' r ‘- 1‘ r: I“ I: '~ - ~‘.‘ A ,. . ., , an ~.,,.‘ ~ .:‘ . I nour, one teen ullOLC& to stlne Ior ,, hours. lie oiltu- “we lilter— 1. l. ' - . 1- . . ., 1, .1 ,. ° ’ c' 7% t. 4- fl ed 11t1 siction and tuoroadhlg IQSne' .1tn 750 ml. 01 'q percent bloo- a ‘ _ f‘ ' ‘ "’ " 1 "L r‘ rs 4? -. 2e cgsein to- returnco to toe Lottl:, “no ti er liter oi so percent , A . 3 , - . .vr -~-r . n,— rjv - n q «a A 'v-n a 0-. ,: . ~ —‘ alconol aLLCu, an“ it yes cglin sullen 101 1/2 “our. Alter stunt/11Q ul. o1 03 7e r cent alco— |-’) (J *‘S H k.) LY D a“. '1 S p... (I‘- I H) P" 1.4 fl ’1) ' 1 7) _ l ' :7 (1‘1 , J CD {:31 Ho (0' ‘4 «4 \ A. ( ) hol, and finallv with 1/? liter of WC fierce.nt (by wei jnt) to fccilit ate .1 Co citrate. L POL 3‘,.:7 Torts fl ( FA ) "\H «1-1 ml- vd. Mr) V}. r). “L 1.1.). c.) g— (- KnHPOy :13, 3 L l K01 125.29 3:01 77.M1 0,. ( nrvdrous ) 73.53 H ('1 , fl . ‘- ‘. ~a,‘9V ‘1' :9 Citrate 1” ion 9w. 0 4 J J .s‘\ (Q B L? () {f ’4 $ P N '- V V V \/ VV V V \J H K x! O 'Urvl C) \krj f“. v 17 K2111 :(SO‘ )‘L 0"." :JCJ'. L \ o 'L'N -4 XI 0 0 KS! () f\-"." -' ‘ -'." ‘+fl VVVQJV 54L~1 U00 F4 \H ' ~' . o‘- I“. ‘, . . ' . .fl '5‘ a 1‘ " ov-~ 4 "3‘ ~' A . '7 r‘ r “- i3? melugea ouantzties o- tn? colts Lure thorond;lv r140; en: E roune in a tall will. Owing to the eli'htly higher content oi xeter I L‘ of crystallization h.5 percent of this fiixture 3&3 used in tlcce Oi L‘~ -: ‘-.s 1‘ 11" - I.» 1“: A: L -‘.~"" Vue OTL~1nfil Csoorn-hsnuel e;lt niutire. l‘ ' ‘2 +1” (9 ’1 0 e L} d O ,4) ”La ‘—- ~ -- - :~ " ‘x, - .. .-,-- . inc QLJJCioVeu Least Tie (rowers; (l, irofi cmuer's Jemgt “Jifix ‘-F, ‘ ‘1 v“ r‘ fl. 6 V ',‘|. a,‘ t,\ 1‘ "‘II'\‘ 5% -‘. ‘ v- t.- -|.‘. C‘ ‘vv(\ a y» - ,—\ n - ‘ “at U88“ u-.l Li- 1...». L--l‘\ QA'-Jk.l«sy~. 1‘1. 81.7.00 - Tuag'v‘? .'.‘;..;3 .lche 01 can. IO-L:;,. Q ‘- ~v .~- (‘ . a ‘1 fl . ' 1" Av, . ‘- . ‘ . we -" . W A. \ ‘ r~ A maler's decot chi v.1 m buU” solution (Uglnq ror 930$ l_o OTJHS or P ““-J~ wh“.u 0. r fl M l) EOQflub gLHSLQre la a thSoure COOAQP. tralized with an equivalent amount of standard 331. It was iried in rw - '5 AL -."\ r ‘ .. ‘3‘, uh cveg e; Lu C. gnu regroinc. J- - i‘- ° :- . z“ -. .. .- .L ‘7 4- A V‘ :“o la Lne ore aretion oi tne sll-ite tr .Vel .‘ ct \-,/, .0 Met / .. .0 '1 ° - 1 . ., - A I - .. - ' .- 5/" . . -. 4 . ‘ L4- -_ QT'VTS C i C-I’1*‘CL :__r)l“1)‘_‘1.".i U ‘ I‘ ‘~ "‘6 ‘-' , 121 0. /-c:\ ”L... \. LLD3 "k‘lu L [.0 u ult: \ , a .. Q Q Q . ‘ ‘- -. ~ .- 1 ~u -L . -- V ' ar- n,‘ "1‘ ‘ . L-- 4" —‘ ‘.‘ -- - tnlOulOO tyre“... u.til a ti Cl no.1“,s r aC“-. -;~ oo.t_r =.is -t0719f- d riK ‘ r' --+' 1 0 (‘Ir‘ 1" (\L ‘1! K -1 -—1 \(‘rr‘ +q1 a l’fi—1 P\ 4‘ 1 .— r1q~r~ [Nt‘h 6 g'lllj». $.- J in Abv --mr-- 91.. dd”. LC .. J|A1\_ \L‘J U] L). j \ fin to ' L... n ‘ V , 3.; , 1 ~ - - ° 9' . - - .( . cort;noo c- the Lo.tle L_s tzen (riec L urlilefi escln e a tCl‘C‘u- r‘,- . _‘L . W‘ . ‘J— M ..."J.‘., : - L' .. t'- . twr: n,u erceeuin _~ u. no tu? no cl t 0 re letion weriow t-1_ sul- Q .-,“‘: ‘7‘ r‘A-v-r, r|t ‘1 11t r‘fi“ ;‘.-‘v 11V, 9:11: ‘d 1. f3 i"“j$va. ‘vI 1"";7‘ pu‘,+ ‘ ‘I.-L.L(‘-4A 9.9-: :_.r--- C!“ O\.‘ ", —L"~L Al-Lk. bA‘r .L 3-,... -Lr1‘ .n L~ 1 us; .A;l-: np.-’ 5‘1... 1. . .-UC4A S cod liver Oil and Argo's co-n:trrc1 were u ea. 0 f3 H ‘0 (f 1...! "I D ’1 O ’1 :3 O (P Cf 13 ab ’3 N ill (‘\ 3 m 3 t e N (7 p.00 3 ’J H. P!) H. a 1 ,5 d‘ :01 ‘_ r‘ O O I V I ‘- 0 fl lo Oulfllu inforeetion VAL fi ‘4 _,‘ . ‘ ',\ - ‘- ,\ -. ‘1‘ -.~ gt OI tn? anim.le on ice VLT101S Sujyle- ments an analveis of variance was ma'e accordin: to onedecor (w ,. F4 ( 0 Ovaf: c 7-.f\.-S‘\ p A .0- ‘r‘ + ‘ ‘O (‘7‘ r1.u-1vo~=c. A? 7ft" .ry'a ‘y‘mflrw‘ c a t‘jfi -:\P)'A"~ 0"- ML}. |_.L..'- V0 LIL; J (Lu... CV4...~.L', .‘_ A g V .-. ‘¢ girl-«BAA JG 0‘; K. \—.Lluo 0.; :.'._, :{.l 1.9(4 p .‘L \~ \ 1 r‘ 1 ..‘---‘ fi‘-. --. .“fl: ’ 1.5‘H‘i ‘~ . . r litter contzoi--ewce or: ;1 ptluv clr591-ie1 accori e, to toe two inle— ‘ 1 : ‘1‘ q.-.‘ .- - -‘ q . ‘- \ ‘- . 'yF-c -~ 7 flr‘ vr‘ . percent cr-ter1u (suwplenent: nxn litters). in fnls Leg tne M~in wiri- 1“." 9 ' 1' . '~ - 'Y, .-. - -,. _,....-‘ .9 u ,. :. - - etion oi cli 4“ unitals CDulfi :u uoeo lhfittuu oi tie tarihtion mitoin - -n r'" 1 .— : ,t 4. ' ' u l- ° .. [- 1- .. . r one Oren.» - 1~ x'wnte 01 v Jul tion is retJPq; .<:1.; tye .v~: r~i i 11r— , u .a -n: ~. - h- . -~ I i ,. ~ « ._ a - e“ unmrer an; tflJE a true~ jvfln. also. Li 0 brouos coulc we inve;t1— Q.” ‘fi " 4— a 1-»-«1 a. u‘ u, r, v ' . . .7 . l‘ .5 “ ‘- LLB tetxoeh rho iemcnt Drones. -30 r;mainin5 variatioi xiv Le observeo J H 1". -'. ' ’ .M 1" 1‘ '1 r 1 w . -‘ . v - ~ . n ‘ 9- . .. ~~ o oroolen Kai, ticn, to mgie a valin GatlflathL o: tn; elmerimentel error “*1” f f (1 fitF‘ Yi‘iT'ID ‘ravv Y'vs'avf.” fl ’1‘ . f "N 1+1 '3 ‘5' ‘5 (—3 ""37‘D O . "" . "p. "' L " 1.. L \J. - J L", ._ r. n ‘.~J ‘JJ a-A\. “Laio 0 A. l J \n._ - 4.1,: r L’ALK r ..4 I.‘. i. '1 01 - ..l i 1C. n»! (- 1 .1 *grmio‘V‘e‘. 'r- - 7'1 u ‘4. ~ 1 n vgfite (: = .m.... , i313 : Vilne rerfl irow stucent 5 tan e dives error ¢~ . (1 . f‘ . "fl O'C‘A ,' 1- - . ‘ . ' ‘5'] 3 -“‘ . r- -- r: . 9-: | ~ u». ‘N - . H r‘ ‘ on lnulcqthu ”Aether “Jere is a si,uiiicAu milierence Let een tge melee. 4-‘A .‘n- -,‘*~- ‘ ‘wv t“; 3 _n v-w'l-l 0“ t I ‘ V -, ‘ rF‘Wflfi 1 fl] 1- Ov;.er £2173.ch u. (A, ‘31“..t'hjum - : ' - . -.i::;O 79. has -.. ..- a... 4. *1 .r‘ x ' '4.-. A A' M °": were also loohtd u“ in two o¢ e taole ior orcoabilita oz :lvulliCaneS. “'--I u r. v if" "I -«- v r 1 " J" 'A- v" -‘ vcv ~ -\ ;‘ r- , ~ r .« ww; L bum Jr}, all tue test groufie “ere Controll & so the : flr‘ ‘ ‘ . ,.\ ‘ — .Qfi‘, ‘fl 1" ‘5 . l. LJCQ .”Oup nod one aninol iron eccd litter. ‘ ,.. _‘ ‘ :4 LR‘- I v AF-x “ O :Q r n‘ am _|O~ 0 _~ 1‘ 3. Eaca Group ham Ln" sewe mac“ JClbuo at toe oevinnin; of the 0 test period. h. Each animal receivei M gamma thiamin chlorite cm.h do? incluulng 1? 3V8?- Sunday. 5. The food cozsumptio“ nos controlled within litters so the age food co sumjtion of eech group would he about eeual. o. The brsal diet figs tFe some for all groups except grouy 6 which recoivef sulfite treated yeast in place of autoclerci yeast in the basal diet. 7. Th? only Viri'tinn can he: the ,1Citisn of tho various vitamin E surplomerts to deterrine which 0795 are low or lrcfiin; in the Sher— ”“ase diet for CSScyin; "itafiin ?l. Ja.) , O gesnlts ado Dismission .. 4.1 ' --- 3., tax. (m 4.‘ ‘ ‘ILI .._°.-,-z: ., o i I .-1 . .. r' 1“ Id v.':1: (1‘5- ‘4“; r" ‘J -Jr \ HL -1‘U',r.llstg' K14J$L 0.1. t.-.) u'-e'~.1-LO C" O; ti.e ~11 -: A _ A o ‘ ' o _ f" a ,. J. a - v ‘ ‘ n‘ “A A sq o'er"un-Cna;e Vit bin 1 Lul1c3~rt fie! in! axcru 9 \e1 wt or tne a" L ‘7‘ "\ u 1 ,N v . “L" . - ~ I ,1 r‘ h‘: s‘ v c v‘ v- I” : A q r Us h.:~ ;lacel on tie bhlfl -H- rce 1et a a} do‘“ oi a e var-~1 3 7‘ r‘fi (”1 0 q q o l . ~ ‘ - a l“ ‘ *””W ‘3 to y; arams. In? aninale were oerleten on litter“ .n« 'e ° :3 , .°,.:! 9 , l 4. f? 1 . 3 ~ 'LL- ,. . ”I," I 'y. 4.‘. ' peric; varieo 1r~m ll b0 14 .ajs ior eacn llbu r, at anion tine their ' ' "“1; ~ vr‘v': «A 11‘“"‘M “T: t .f'. . raw Y1? r r w unp' Jot ~ T" r‘.'- ‘m an‘fl’u‘\] O “91:21:. Q w“.-- LL11, 4.1 U“, ‘1'} O V‘J‘ HI‘ Knit. $.19 grieIL‘l'l “441;”- L;_ ”INC“ .:.A 2;. ”1'" P‘~ V ‘ ‘w ‘ ‘z‘ "'— 1' r‘j’.‘ . r~ v‘. I ‘ 4", ~ ~ A I‘ . .n I‘ . meat VrJto ra‘-»t Iron 3,.7 Jo ;U.9 Ur.L2. In tde ngéin01; are ce— consumjtion and ;rovt3 recorfis c+ ( \ t J. ’4‘ "D (.1 :‘f' )1 l". '4 J [.0 I’\ »2 (I . J ‘- H O H I (-4 i H \J "2 F?) c?- _“ "J F‘) 0 \_J LU O i” ‘) ‘ t; }.J P .— >1. 3—) *4 (1 O ’- ”D n- \J C.) f—‘l‘ S I) f) D E; *‘5 O t—J .J 0 pl ’3 a + :3. Ho {‘5 i .J 0.1. :1- C+ Cr. C I" ;V D *‘5 'D «9 (‘5 ”I 'D {-7338 017. ’ " ' - ‘ 4““ ‘ 'v A . - ,r‘ ’1 l .1 ,-. n~ ‘- v\ r‘. . ~ 4- 15 same aninalg ”we ate all tkeir loou ”nu gone t;ot alc not. . Q- 1‘ V_ _ 1 ,- _ O O . 'A O _‘ +- 5“ ‘ ’ . 1 v‘ ‘ . 'Iq ‘- It Culrt I, tne nern QLIK 1L _e1,1t oi eacn amnrleuent ”road Ior " '0 "‘ P‘- ' -‘ n a . .‘ -. fl" 1 4- F.“ ‘L p: tr ‘-~ . ‘ . o ' ‘. tfip 4OUF neu:€ test oeiiou is eat 2. Ice rat; JulCJ receivel onIJ ‘.° .A.- ‘1 -* mu '. ."H+‘ .- °r v‘ ,L;«' .~ :3 H... til.uln calorlce or LEO 11 tagiulcn IHCQIVFU uicatlnjc aci.. ritl— “'1 ' 7'. "1 "1 1 1W” ‘1 t‘fio "'3'“ "n fiv"? .'--n nvt‘ “ T 5 r;- {-7 ‘wf‘w 74 17 . w .L...;iv 11., C1,... 14,le f._ ;;<4...| wont... fl . p.01 .rl Cary/'3, \VncL !.'.J.a “‘41 \ L“ "2 ( Ht 0.. 4-2. ‘ ',‘ ‘ n H.-- P. um . _- ‘ b 1 0‘- ,- oi g2,jo Jrque Icr t e tel-filn oral , aa.+a ére.s Ior tte rlooflaVIH , rm w‘ ,n- q hi '—- v- ‘- ‘ . ‘i . . I ‘ . 1 ~o \w I“ A Qrou¢, lna 53.x} arses for tie KlCtulnlC ac1a Oriip are clooe. " "' ‘ I. r‘ . -‘ A . . -s O n‘ . . I 4 . "1' 'I V“ T?! . ;r0utn of tne £41311: receiv1ng crystalline Vitanin Ir ano tne lactor ." “ d I" . "‘A '1 I“ . r- I -’ “.9 1' +‘nfi . v a - A #1 - It ‘I J“ ‘ a h piepa ation snotca an increaseo dromth taut ls Jarz~alg bleat;r tflcfl 7 tne other Jrowth curves, Y3ile to ee on ”sulfited ve st" sh ran some . m . . ‘ 'N‘ . ‘-"\ r- . ‘ - . '1 -- PA fir“) 5' A ‘n I a . vmcfi increase. iheir finel Jal”: 1n avlvlt were L9.cl brnms Ior the an fills receivinv vitamin Br, 10.2? grams for those receiving Factor “ pre- 0 ’— .\ naration, and 26.1; for the "sulfite yeast" group. Table VII summarized the mean value of the sunrlement groups, each 1" of which consisted Ci 1% rats, one animal from each litter. There was S ”e variation in the Iood consnnption Between the Q1; 0.4 , IL .'-n ‘ ‘ 1 elent test r)1' aye Average gain in weight in grams 19 CHART I. GAIN IN WEIGHT CURVE OF RATS 011,6 SUPPLEMENTS 30 «— Supplement groups: Thiamin chloride —— Nicotinic acid 4 Riboflavin Vitamin B ..... - Factor W preparation- _ .. "Sulfite Yeast" basal .— 25 20 15 10 5 0 i 2 5 Time in weeks AA “U .fionommo doom mm» Had one macaw node ca mama exp weep on» mo mepCmopew one mpncmmnemn w ma weapon mpqpomm \I. \. .1 . I4 0 o .00 00 000-0 .1 w wm hem .o u.crfi m;.om H~.mm 2H ... . . . .. ga>eflt0lam \. N I. o\. x: \.I. \ 0000... o o. o :J . it .1 u muon.o H UHH :u.«o ma.ou :H . . . . .eaoc carapcoafl emmfl.o man mm.mm mo.wm :H ..............eeatOHmo naeaage .mfim .mfim .mfim. w conmm CampMEScho muoos 2 unawoa madaflud §m.fip doom fimnm voow swam wapfinfi mo muqoamaamdm on pm Mon cfimw omdnm>< omoam>< omMpo>< 909852 mw¢m mo homamUSmhno moon QH4 wnhumw mo «Ham Mu methm Hwb mqmd h .A. .3 l & ..-\ wk... by V l" " 0 Th '1 ‘~ t k) 1 ,, Pu o '21. } tile + .1 C) I A gition or 1 1311 . I wirot .. , uni «‘1- . \.v.. . «3 4‘ ~ r‘.r 05V 7" Ugo Y q KN ater rowt I'I‘ ‘J L.‘ I r6001. 08‘? . r) -.. Quit '0 $— ‘ul 9 f M ice lDt "y A» S .1 fining... I very éram g ‘or e eeent ‘7‘ .L p" II r 'x.’ u . "‘ C" ‘1“? AL 01 Leak/Le '1 q ' v .A. v... Q .1. .1. +v 3 - tood 1 i A 1;. vi.'_'. .I' 11 4- . 96 ~21 "‘1 5'. »,“'~“d.- 0" OJ. .9 Lu) ‘- :rcert .4 1“. E. fl... 0‘ . ‘- lb'n du “»+ .4- _,L, U OT' rvi( yx 'I '7’ d 'actor .l.‘ ., if “Q L C€ i‘dr l 7"- .5 .A .1 r! ./ ‘ ‘ r w— ‘ . » 0'9,pr tor '3. v.3"; -—W hac .1 l a r- ‘ '10 we .A 1. & Us...~t4._.. 1‘ ,~. ('57 1") .l ‘l‘ t :e or autoc A t y~~ rcent o '- h “d ‘ ‘4‘. Q n t7 ‘0 e yeast P.."\‘ L. .J 4. .- U as J. ‘4 -I. nl*1 u l w .Yl to ("C ,,i i ”1 ‘A w n 7‘ 0 $1127. -1- l" \- :51.- r.(;\ -\,. a L‘. of o1. LL 34. 1.“ mu .1 H .1..-& "" .‘v S..- tested a i“ ..-r-, \. Infid- ('2 . tIOE ”54“? ‘ V ‘1 l‘ l 3.21 L I"? .1; .. V 3. H w;"' 9 Q v‘. . . Q \ IIII t 4..! 341 ‘1‘; \I \JHJ .)IO\JI.I‘I-I.‘IIHM.I1 . phyla! 34 \I. Ch ‘I I1- 33 l J\ i1...l eunu‘ HOkV.c.-fi +(r8 (H01? U .fi‘ (. +FPIP‘. W3. "Fri. Wm“ Hrdakl; r +(MQ.IVOFC(.: - |+x~ prp...v WC. WC. ..Ar...l.** AL A); o ., a - :li - - . .. .\ c .s mag H can»: claqrrn "we ra‘oarl I w-4.x JJ-44 w. 4. .y -.-:s 1H“ -3 \ *uc;r p (tr r UL r;; rHH Htawg. s.x+«z 1.) {VI _))4.\u ) Jaw \KJer _ z 1)) JJI IJN .HU ..‘J1 'J)‘-‘3 0L. 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Ho: CHFF‘UrIVHH CHfiIM-u H.P.:.4UH J. flJ _..J.‘) ;N1\ U)\JJ_. t: O r. . . . r . . L. r N... .r-.\ (PM U. H:\41F .qolfl noJII‘NUN I. .1 \ IIIOAJ .< I a-u))1 \J ‘ 1U 4\JJI¢.' \ )4 v «J- r. .....L r :4: :r...: I 0) Call u x.\ (If u\(fi‘ Hr) m frrsvvcu .uwf. .rtflutn .LV .,.4 - :3 D . I .- mondrtca iauomyumq we (on. 0 wooden o If P\ I C .4\ f \ lul‘,yilllz“ lulli. I. -II‘.‘ I I‘ .I.in|t!-’ ~..I.II. - ‘. 0......3‘4: {4‘ .c OMJ. 1) XI). 4..: I. I. InlIIJi.d rlvl.\'vltl_n -Iar Luv-H. c F’\ p‘?’£*(4‘ll‘l ru)\ '0’ It." I.‘ HHH> qufl +- , 1~ -.. ”,— 1 1 v- 'm. '4‘- ‘.‘.4- a ”A. 1. - .clos cedoic txe 1. level, l4xlcit1n_ ttat tue v I :t1;33 1? ‘y\ t“"‘.“‘.\ a ‘ #40 I‘ p 1 ‘1 f, a‘]"\ .1 ." ~'- u\)‘ ’9 n p- ‘(3 3‘.“ a .~~1’ he .I', -- o»)... . UL ~..; bA' .‘ .. :1~JD p, \n. L" 11;}: fissit've ircltase fror cone factor adiod to the diet. The ”t" values ‘be*\'eC“w t we t‘ 1' ‘1r r~»nt1*:l (5r7m1fi -*"' fr c“ oi‘ 3‘»4 otla-r "r:1r“s (Llon the Drou;; witb Vltfl'll :5 ard Factor I vreparaticn Xa‘ values t t mere siv-ifi";ntl" differett, to‘4Q Ureat'r ttan t“c 13 value for 12 ‘ained tna. :mch by c‘nonce. The increase of the "suliite yeast" over tke control may have teen die to chance errors. Thus, the ads: ion of o: Eactorfi “repa ation Sififlifld a necessarv ”rowtn stinulant lackin in the autoclaveo veast. In Teble IX the anm ysis of variance was leternined on the gain yer gram of food eaten, which elirinates tho factor of dif‘. erences in food consunvtion. The ”F" values and ”t" values snow the same signi— ficance as in Tabl eVII I, which in i cates further that vitamin B» ano Factor W or'o'ration not only stinu ated better on but oetter ut l— ization of food. It is evident iron tiese results tart one c~n make the following 1| Th- alivals v1ic.o rece ive ed 4 ganna of txzi l?”ln cmxlor fie cn-v one S I '1. H. 0‘ U I L‘- " IIq .‘- ‘0 Q on -"\ ~_ ' o_ ~_1 o . ._ A «.-.4 LJObC tist receiveu I gamma -1 tnianln onlorlue plus 1, Damn . .. _ .l'. _. ‘h . F! . ‘ fr 0 - "v , "r I‘ ’7“, Q flavin had the same avela e Gain 0: 2.a glans in four “eels. -nns, ‘ .‘ 7 : ' , .C‘ . .f‘ 1‘9 . ‘- p 3 - 03‘ ~.. -— ‘ 0 2‘ I a v '1‘ . ‘ tne accltiun oi riboilalin naa no ei-ect on Urea.» Cl the ani:i s .. .1cn seems to inlicnte that there is nro icient ril flavi- nresent in the autoclaved yeast. a}, ’ F‘ v ‘ A -. r. 3- 'a " -. .- m.- .A., p‘ . ‘. . , it over tnose rCCeivin + QCMLM tmld.Hll c1 loriue only, . "x : -a 1 r- ‘I‘ A ~ r a ‘ . ‘- . 4. r . ~ . a ‘ <. 1 flogtlné t1nt ”1L3 r tne rnts 10 not nee' nicotirio AClQ 1n tneir ‘. O ‘ :0 O ‘ I Q“ 1‘ ‘ 'I. L. O ' :- niet or t1.t tzere 1s snf-1c1ent 1n t,e onsnl oiet to Feet oneir 1etd— , The ro1 , whose basal iiet corte'nefl snlfite treated yeast in nlece of autoclavei yeast showed increase in sin in wei ht. Hcvcver r- . . n v“ .H- sv‘. ' 5‘ . ' r1~ -‘ - r- ) . \ .-', , he st1tlst-Cnl Annl.s1s 01 t1? 't" val1e (Tntle VIII eni IA) snoved that the gein in weiJht was witnin normal limits of experiments.l error of the assay method. It .0 .1d therefor seem that the choice oetween usin; ‘utoclaved or sulfite treated yeast in the basal ‘iet would de- .- " a .- . ..° s“ 1' ”“1 . 1° .5" ‘ pane on contenlence or role at1o n uni uwon a olfilotlt countered. All excejat 3 animals in this stufly on th basal diet using ulfite venvt dovelooel diarrhea. Cne nnimnl diefl as a result of the cf ,_. J (D . r‘ - r‘ h '7’: ‘ . c .A - C. W . I.“ h ‘ . V a“. ierr1ea. 1ven “its one interference oi , alarlnes, t21e anlnlls thi 1n ex:erimentol error the same as the animals receiving the (:31 M f) (D {.11 +1. ideal diet \it 1 autoclaved yeast so the yeast may have been a setter LA - 0 ‘ r A ‘n u... ‘ . .- -- ‘-. - .L‘. ,1. so1rce oi tne Or routh factoro tflnn \as the autoclnved Je&3t, or .ie ~n1— Wnls wouli have geined less emna 01 crvstelline vit-rxin E6 (nvridonine) was adieé C) ' ' v y 3.71811 10 there 15.2.5 an increased growth of 3.1-.) 312.123 above tLIOSL. receiving 111;: thiamin c loride supple em3nt. This was great enough gain over the con- trol not to be Cue to normal variation in growth alone. Hhen tested stotisticoll‘r the ”t” v0 ml 1e was greeter than the "t" value at the 1 percent level. This indicated that the vitamin B Optincl frowt1, and t21€t it w=s present in subootimal amounts in the autoclaved chloride, 3.7 . f‘ .‘x 3"? Circ‘h "I o.» -’. ~.b K": I' :3. weight vLen the- , a O for rat include i? or Pu 33 I‘O‘L’ V L1 1‘ a. n $- u ‘3 LL 5 is V. eived 4 rr "rams OI :actor a “a D 5‘ 'nnv ' 2.1;. L‘ .A ‘ ~ ' -- . . "- hue draftest meal hm 1; sucscgnce l; -..+ .qflt Wm”) :‘ib‘JIg‘§L’I O +.;‘4 0 v... . ion .- ,- 4- }: I") pv 1“... v the pnfioclavc O r- v-1 1 75 MA...‘ a L t. v: A. yd. by the I.‘ q I: (.— 5 39d v. W‘ I' ~ ‘- lgbs \zCI‘fif v0 R Q ,1 and th R.‘ ,. 0;.01‘Liélll-L ’1 11 (.A.. 33833....”11‘9. c-v- eports in ":9 .mV "'3 we}. Ho elm; se a DJ ontains Factcr W l-controllc 1")!“ and so CC uld L:ve been SOLITC‘B ‘ 'iet to the h *‘P‘yfiw-II“ L; ... , -. u.)"" ‘ " fl- '..' - - :uctor n Lre;0 (Jig: P. t 4.x the nreparation {1 H. m ’t'I1C “’33:. l‘ ~,'~ k~¢ «I1 . 1 59.1119“ :“Ct thct tfie i' fiIngfl 0 th9 diet. T‘ factors 4.4 acfl- heretofore been 1331 7733 be ruiha for the of thiamin in foods. ’37 ints boo L‘b‘ . 1 3'1 ‘ l“ v ctor ' I Lrv l [3113 i0 nan-C. 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U W- , J ._ .x (. .'_ . .. . .1 J" ' - J; A; J. o A O O J-‘-0 -L--‘L--’ f. b“, ‘- "/.,.//. ‘ - M * fix - . I w: ' ‘ . “ ‘ H ‘ ‘I‘ ‘ ‘ . V ‘ _.) ..‘ r\rr .1-1~\.«1 all t .‘f' I‘lq'Dy ‘1 .‘J~O~p‘fi ‘f1.v\lo 1 ;+:. T“ L 14:} ‘v it'fi.n1:L ; o A La _ vv 3 \ . - '~+ ‘ ,. " II . r— Al,“ / fl \ .- - . - q I .1 - t 'L ‘ C I k‘ ‘ “ “Vt- t \ L r0. " 1‘~1-'. J V J. .«O ’ Ir \ .0. :1 a J.— |\ 1 -’ t . w- -: ~-. A ‘ n: .n! :.~ 2 +n r. «.1 '» ~ ‘r\'.~j L 0 .A. g -—o L. H‘SCT, FL rqu‘lfi. ClutLJ‘L C—L l tn V: 'P1 r)- ‘ f L - . .. —n. — ~. (- awn w ~* 1‘7‘1 h n . r‘ - ..l L r . Du 10 .L -. 1 , J. [a] ‘1 /*/° C“ .1 "\ "V f"..- A. 1 [J— ' A» ‘ J. - . .: ‘ .f‘ ' fl -‘1-_. .3 U;.‘?'.~ ‘CO.’ 1 j. No , \IL..1“».1-‘_:.‘J 1.011 s."'-“L 1 i'l..CI°‘|I‘Pt£/. -th O... lttu'..-_,v'- 84— r . ‘ --‘ ‘ \vv .4 “A“ '1. .— . «r! A, . (1 A:‘_“‘.". ',<:'1':;.C:' (“f \JVr'l L ZUC‘P. '1’).Ll~'7 1.» {3 79:39.33, I’.)‘..CL \J-LIJbr-lo \ .~‘.x_. TABLE I DATA OF GAIN IN WEIGHT AND FOOD CONSUMPTION IN CONTROL (ECU? OF RATS v 11. Growth L Gain of the Average 0 whole groug Total Rat number 1c? 1067? 174 259 513 3907’ 459 658’ 6667 729 789 919 9569 1066” Total Average Weight Initial weight 61 61 65 56 58 51 58 65 57 59 50 49 55 56 797 56.95 56.95 155 week 14 8 7 7 7 12 9 ' 0 9 7 4‘ 4 11 7 106 7.64 64.50 .33 2nd. week 10 -2 0 7 5 0 1 10 -1 4 9 11 44 7 57 4.07 68.57 5‘53?) 3rd. week 5 5 11 -1 4 4 4 5 6 5 0 ' 5 -4 2 51 5.64 72.25 4th week 7 7 6 9 11 1 6 11 1o 8 2 ' 8 8 5 99 7.07 79.50 Total Gain 56 18 24 22 27 17 20 26 24 24 15 28 11 21 ) 515 22.56 B. Food Consumption ‘ Consumption of whole group Rat number 164 106” 173— 252 519r 598’ 452 633‘ 660’" 722 782 912— 956’? 1068'To‘te1 Average 181; week 56 51 25 29 56 27 29 26 50 25 25 24 25 24 592 28.07 .53 2nd week 52 54 25 29 56 26 28 50 26 26 26 28 24 24 592 28.00 g 3 5rd week 54 29 52 28 55 26 27 29 51 24 21 5o 28 28 402 28.79 § 4511 week 41 29 55 52. 55 24 29 56 52 51 25 50 55 28 458 51.56 Total food. I consumption 143 123 113 118 142 103 113 121 119.106 97 112 110 104 1624 116.0 L A TABLE II 35’ DATA OF GAIN IN WEIGHT AND FOOD CONSUMPTION IN NICOTINIO ACID GROUP OF RATS A. Growth Gain of the Average whole group Total Est number 56“ 113 2067‘ 269 526'7 409 506" 609 6807' 756’7 799 849 996‘1019 “53121 weren’t?” Weight Initial weight 59 55 65 60 58 52 65 59 62 62 49 50 52 51 795 - 56.79 56.79 .., 181; week 15 5 7 7 14 8 8 -1 4 8 8 2 5 5 91 6.50 65.29 33 2nd week 5 -1 8 5 6 4 -5 4 10 5 4 9 6 6 66 4.71 68.00 g5) 3rd week 7 5 4 5 12 8 15 00 1 9 2 7 4 5 80' 5.71 75.71 1- 4th week 4 4 9 8 15 o 19 6 5 4 1 5 15 5 94 6.71 80.42 Total Gain 51 11 28 25 47 20 57 9 18 26 15 21 26 19 551 25.65 B. Food Consumption Consumption of whole group But number 56‘ 113 2087 263 525fl 409 500” 603 683‘ 753‘ 793 843 998‘1013‘56861 Avere’ge 8 1st week 54 27 21 50 52 27 29 25 29 28 26 24 26 21 579 I 27.21 5%. 2nd week 28 54 25 29 55 25 52 30 27 27 27 28 26 24 595 28.07 gg 5rd week 51 26 51 27 55 26 28 26 51 28 24 5o 51 24 598 28.51 3 4th week 40 25 55 55 56 29 59 28. 29 54 25 5o 54 27 442 51.5 —_10t81 food consumption 155 112 108 119 156 107 128 109 116 117 102 112 117 96 1612 115.14 TABLE III 3b DATA OF GAIN IN WEIGHT AND FOOD CONSUMPTION IN RIBOFLAVIN (3017.? OF BATS A. Growth Gain of the Average 0 o o o ‘__ whole group Total Rat number so2 148’ 193 283 569 4267 49c?I 594 65» 74o7 80-0 8807’ 94-. 1056” Tofaf “Average Weight In1t1a1 weight 60 57 57 55 62 45 65 54 61 57 49 56 5o 54 780 55.71 55.71 13’s week 14 6 2 11 9 15 4 6 4 11 10 -1 6 10 107 7.64 65.55 .33 2noweek 5 -5 4 2 5 4 6 8 11 5 6 8 6 6 71 5.07 68.42 0 §%3rdweek 7 6 o -1 5 -5 8 o o 9 5 10 5 2 51 5.64 72.06 4th week 2 2 9 5 10 5 4 10 6 10 -1 6 11 10 85 6.07 78.15 Total mm 28 11 15 15 27 19 22 24 21 55 20 25 26 28 514 22.45 B. Food Consumption Consumption of whole grong: Rat number 6o" 146’1 199 289 563’ 426? 49:? 593 659 746' 803 88& 948 1056755551 Average 2 let week 54 25 20 50 50 27 26 50 51 29 26 24 27 25 582 27.5 0 how-0 3.15% 2nd week 27 54 25 50 56 26 50 51 28 27 28 28 26 24 598 28.45 0 ’g 3rd. week 54 26 24 28 52 26 28 29 55 28 24 50 51 29 402 28.86 0 4th week 41 25 54 24 55 28 52 55 52 55 22 52 55 51 455 51.07 Total food . consumption 156 110 101 112 151 107 116 125 124 117 100 114 117 107 1617 115.50 TABLE IV DATA OF GAIN IN'WEIGHT.AND FOOD CONSUMPTION INVITAMINB6 GROUP OF RATS A. Growth Gain of the Average Rat number 55'7 156* 183 243 559 416? 47o?7 626'7 690” 753 819 89o” 959 1049 Tofiglehgggpage $121111: Initial weight 65 59 64 55 58 55 48 55 62 58 48 52 56 .54 785 56.07 56.07 let week 11 8 10 10 22 14 25 6 9 12 9 9 1 4 148 10.57 66.64 g3 2nd week 11 ~5 9 6 2 2 5 12 7 4 9 7 7 11 89 6.56 75.00 g 90 3rd week 1 11 7 3 12 «1 O 2 5 2 6 11 2 O 61 4.36 77.36 4th week 4 9 5 5 6 10 17 11 5 10 5 6 12 12 111 7.95 85.29 Total Gain 27 25 51 22 42 25 45 51 24 28 27 55 22 27 409 29.21 3. Food Consumption Consumption of whole grou Rat number 505 15a“ 1853 249 559 4133‘7 47o" 6262 696? 759 819 896? 959 1049 m a let week 56 51 24 55 58 28 29 28 51 29 26 25 27 24 407 29.07 .33.: 2nd week 50 54 29 51 58 26 55 52 28 27 29 28 24 24 415 29.05 E; 5rd week 55 54 56 26 56 25 28 29 54 27 24 55 51 50 426 50.45 8 4th week 59 29 55 54 56 51 59 55 55 7 55 27 54 54 55 472 55.86 irate} food I I“ consumption 158 128 122 124 148 110 129 124 126 118 106 118 116 111 1718 122.80 3'1 TABLE V DATA OF GAIN IN WEIGHT AND FOOD CONSUMPTION IN FACTOR W PREPARATION GROUP OF.RATS A. Growth Gain of the Average Rat number 76" 15a“ 253 29c? 579 459 460“ 589 706‘ 769 8267' 869 gamers-5.312319 $23338 $21331; Initial weight 60 58 58 58 65 49 48 51 60 50 64 50 57 55 781 55.79 55.79 L let week 12 5 15 14 14 9 18 7 10 10 -2 4 15 6 155 w 9.50 65.29 33% 2nd week 12 2 12 9 11 2 7 16 5 9 12 10 1 10 118 8.42 75.71 g m 3rd week 2 5 11 5 5 -2 12 -2 2 2 1 8 0 2 51 5.64 77.55 4th week 5 6 1 7 12 11 7 15 4 10 5 8 19 10 121 8.64 85.99 Total Gain 51 18 57 55 42 20 44 56 22 51 16 50 55 28 . 425 50.21 B. Food Consumption Consumption of whole group Rat number 7:71 1561 253 29o? 579 459 466’1 589 706“ 769 826? 869 98631029m‘ let week 56 51W25 55 57 28 29 50 51 29 26 24 50 24 411 29.56 :3 2nd. week 52 54 28 54 58 26 52 55 28 27 29 28 26 24 419 29.95 fig 3rd. week 54 28 56 29 55 26 28 29 54 '26 24 55 55 50 426 50.45 a; 4th week 41 29 55 54 56 51 59 55 55 55 27 56 57 55 479 54.07 Total food consumption 145 122 122 150 147 111 128 127 126 115 106 121 126 111 1755 125.95 38’ TABLE VI DATA OF GAIN IN WEIGHT AND FOOD CONSIIvIPTION IN "SULFITE YEAST" CEIOUP OF RATS A. Growth I Gain of the Average whole group Total Rat number 83 16c? 22o” 509 589 449 486? 619 71‘6"I 779 856‘ 87a” 9753 1006’7"1Tote1 Average Weight Initial weight 55 62 55 59 60 48 65 52 61 49 59 50 51 57 785 55.95 55.95 let week 8 7 15 5 15 8 15 5 6 11 9 6 8 9 125 8.79 64.72 g5 2nd week 2 -5 6 8 5 2 4 5 9 5 6 7 -2 6 56 4.00 68.72 g 4 5rd week 9 5 10 5 9 5 12 1 2 0 1 9 4 5 75 5.56 74.08 4th week 7 5 7 5 10 4 9 16 5' 9 4 9 9 15 112 8.00 82.08 Total Gain 26 12 58 21 59 19 58 25 22 25 20 51 19 55 566 26.14 B. Food Consumption Consumption T of whole group Rat number 83 1607‘ 22o” 503 589 449 486? 619 716’ 779 856‘ 8751 979 10067711313“ Aver-Ego iet week 29 51 25 50 58 28 29 29 51 29 26 24 26 24 597 28.56 5% 2nd week 50 54 28 55 57 26 54 26 28 27 29 28 24 24 408 29.14 {3% 5rd week 55 54 56 29 56 26 28 28 52 26 24 55 52 50 427 50.45 3 4th week 41 29 54 55 54 51 59 55 52 18 27 55 55 54 455 52.56 Total food I consumption 133 128 121 125 145 111 130 118 123 100 106 120 115 112 1687 120.50 ,, . - .- . . .4 a . -.HI 1 ‘ .v-~-~ § D | - Q .. n . o g.‘ . ..,0. a \.w a . . . ...: r . . ...IJ..0.. .9.I0d..0 ..rlle-r’fl.fiu.41~il(vofiy ‘. 0... . . . . 0‘. . d....01...\ ...“... 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