.V6 I C °Py 1 =================================: uiNiix^J STATES PUBLIC HEALTH SERVICE RUPERT BLUE, Surgeon General THE DIETARY DEFICIENCY OF CEREAL FOODS WITH REFERENCE TO THEIR CONTENT IN "ANT1NEURITIC VITAMINE" By CARL VOEGTLIN, G. C. LAKE, and C. N. MYERS u THE GROWTH-PROMOTING PROPERTIES OF FOODS DERIVED FROM CORN AND WHEAT By CARL VOEGTLIN and C. N. MYERS PHOSPHORUS AS AN INDICATOR OF THE "VITAMINE" CONTENT OF CORN AND WHEAT PRODUCTS By CARL VOEGTLIN and C. N. MYERS REPRINT No. 471 FROM THE PUBLIC HEALTH REPORTS May 3, 1918, pp. 647-666 Mat 31, 1918, pp. 843-868 JUNK 7, 191S, pp. 911-917 WASHINGTON GOVERNMENT PRINTING OFFICE 1918 Ponograph ^ ^ <* D* of B. NOV 19 ISlg , The Dietary Deficiency of Cereal Foods with Reference to Their Con- tent in "Antineuritic Vitainine." 1 By Carl Voegtlin, G. C. Lake, and C. N. Myers, Division of Pharmacology, llygienic Laboratory United States Public Health Service, "Washington, D. C. Introduction. — Recent experimental work has called attention to the importance of certain so-called accessory foods or vitamines in their relation to proper nutrition. The work of Stepp, Hopkins, Funk, Osborne and Mendel, McCollum and Davis, and others, has demonstrated that a diet is not complete and does not permit of adequate nutrition unless it contains sufficient amounts of two acces- sory substances. One of these substances is essential in the prevention and cure of beriberi in man or polyneuritis of fowls, and has been termed the "antineuritic vitamine" or the water soluble B. The other substance occurs in certain foods rich in fat (butter, etc.), and has been designated as fat soluble A by McCollum and his coworkers. This paper deals with the occurrence of the antineuritic vitamine in foods derived from wheat or corn. Researches of a similar nature have been made by other investigators, and the fact has been thor- oughly established that all natural cereals contain an abundance of antineuritic vitamine, an amount considerably in excess of that needed when these cereals form the exclusive diet of animals. Valuable as these investigations are in regard to the feeding of animals and the purely scientific aspects of nutrition, the previous in- vestigators have not sufficiently taken into consideration the fact that cereals in the form of the entire grain are only rarely consumed by man. The bulk of the cereal component of the diet of our people is represented by wheat or corn bread. Wheat and corn bread are largely prepared from flour or corn meal which represents only part of the wheat or corn kernel. The commonly used "patent" flour, for instance, contains only about 70 per cent of the wheat from which it is milled. The question, therefore, arises as to whether or not bread made from "white" flour or "highly milled" corn meal includes all the essential food elements contained in the intact grain. The present paper reports a continuation of work done by others with respect to the antineuritic vitamine of corn and wheat products i Reprint from the Public Health Reports, vol. 33, No. 18, May 3, 1918, pp. 647-666 4 V1TAMINE IN CEEEAL POODS. i- used in nutrition. In a succeeding paper certain other dietary attri- butes of these foods will be considered in their relation to growth. Previous work. — Experimental and clinical work on the etiology of beriberi has demonstrated the fact that polished rice may give rise to beriberi in man and to polyneuritis in fowls when used to the more or less complete exclusion of other foods. It has, furthermore, been shown that substitution of unpolished rice for polished rice prevents the occurrence of beriberi and polyneuritis. Moreover, it has been found that the rice polishings contained the antineuritic vitamine, the substance which is responsible for the prevention and cure of beriberi. Thus unquestionable proof was furnished that the "highly milled" (polished or white) rice has been deprived of an essential dietary element resident in the peripheral layers of the intact grain. Because 01 these results with rice, the attention of investigators interested in this field was soon attracted to wheat bread. Thus Hoist (1907), in experimental studies relating to ship beriberi and scurvy, carried out on pigeons a few experiments with rye and wheat bread, baked with yeast or baking powder. The pigeons on wheat bread developed symptoms and nerve degeneration characteristic of polyneuritis. The addition of yeast to the wheat flour seemed to delay the onset of the symptoms, but did not prevent a gradual loss of body weight (30 per cent). The pigeons fed on rye bread or whole wheat bread, however, lived for four months without showing any abnormalities. Hoist does not mention the kind of wheat flour used in his experiments beyond stating that it was "the flour ordinarily used by Norwegian sailors in tropical waters." Simpson and Edie (1911) describe a few additional experiments on the feeding of "whole wheat" bread and "white" bread to pigeons. On the "whole wheat" bread the birds continued active and well, and two pigeons paired and successfully hatched the two eggs. The pigeons fed "white" bread died after an average of 29 days with an average loss of weight of 26 per cent. Typical paralytic symptoms were observed in these animals and marked degenerative changes were found in their peripheral nerves. Several of these birds were treated when extremely weak with either yeast or beans. Great improve- ment in the condition of the birds followed this treatment. The reports of Little (1912 and 1914) concerning his five years' ex- perience with epidemics of beriberi in Newfoundland are very im- portant, as they show that in this locality beriberi was due to a diet limited to "white" bread made from highly milled flour, molasses, a small quantity of fish, and tea. This author furthermore presents evidence to the effect that the substitution of "whole wheat" bread for the "white" bread was followed by a disappearance of the dis- VITAMINE IN CEREAL FOODS. 5., ease, in spite of the fact that the other dietary and hygienic conditions were apparently left unchanged. At Little's suggestion, Ohler (1914) carried out a number of ex- periments with chickens, in order to furnish experimental proof for the contention that a more or less exclusive diet of " white" bread was the chief cause of the occurrence of beriberi in Newfoundland. Fourteen chickens fed on "white" bread, made from " highly milled", flour with or without the addition of yeast, died within 28 to 40 days. Before death the birds exhibited symptoms of polyneuritis, and histological examination of the peripheral nerves revealed consider- able degeneration. Five fowls fed on "whole-wheat" bread and two on whole wheat lived in perfect health for 75 days, when the experiment was discontinued. Similar experiments were carried out by Wellmann and Bass (1913) and by Voegtlin, Siillivan, and Myers (1916). Nature of the present experiments. — Pigeons and chickens were used in these experiments for the reason that they are better adapted for feeding experiments with cereals than other animals. Only full-grown birds were used. The symptoms resulting from a defi- ciency of the diet in antineuritic vitamine are easily recognized in fowls, and curative experiments yield striking results. The obser- vations made on birds were confirmed in experiments on dogs, hogs, and a large number of white mice (these results will be published elsewhere). As a rule, the birds were given as much of the various corn and wheat foods as they cared to eat. In a few experiments, by mistake they did not receive sufficient food, a fact which caused them to lose in body weight (experiments Nos. 18, 20, and 25). The appearance of polyneuritic symptoms was regarded as indicating that the particular food in question was deficient in antineuritic vitamine. The polyneuritic symptoms were always preceded by a gradual loss in body weight. The peripheral nerves of a large number of the birds were examined histologically at the end of the period of observation, and always confirmed the conclusions drawn from clinical observations. Birds exhibiting polyneuritic symptoms dur- ing life always showed marked myelin degeneration of their periph- eral nerves. The corn and wheat products were obtained in the open market and came from various modern roller nulls, old-fashioned buhr nulls and various bakeries. As a rule, the total phosphorus pentoxide content of the dried products was determined, as it had been shown by other investi- gators that the phosphorus and vitamine contents of rice of various degrees of milling run roughly parallel. In order definitely to prove that the symptoms observed in the animals kept on deficient foods were due to a deficient supply of 6 VITAMINE IN CEEEAL FOODS. antineuritic vitamine, some of the pigeons were treated after the onset of the disease with various vitamine preparations (Funk's vitamine fraction) obtained from brewer's yeast, ox liver, or rice polishings. Further evidence was obtained from protective experi- ments in which there was added to the deficient food a small quantity of i vitamine preparation. This material was prepared according to the method described by Seidell (1914). The so-called activated Lloyd's reagent consists of fuller's earth (Lloyd's reagent), which has been treated with autolyzed yeast filtrate. It contains an average of 2.3 per cent of total nitrogen, which is derived from the basic substances of autolyzed yeast filtrate. As it was confirmed in the course of the investigation that white bread was deficient in antineuritic vitamine, a few experiments were made to determine the effect of adding an extract of " whole- wheat" bread to the diet of "white" bread. This extract was pre- pared as follows: Two hundred and eighty grams of " whole- wheat " bread were extracted twice at room temperature with one liter of 50 per cent alcohol containing 25 cubic centimeters of concentrated hydro- chloric acid. The filtered extract was concentrated in vacuo to a small volume, so that 1 cubic centimeter of the extract corresponded to 1 gram of fresh " whole- wheat " bread. The extract was free from fat and contained 11.5 per cent of solids. The animals were kept in wire cages, tap water being supplied daily and grit from time to time. The bread was fed in the form in which it was received from the dealer. Water was added to the flour, which was made into cakes. The cakes were dried at 45° C. and broken up into pieces of suitable size. Throughout this paper the words "highly milled" are meant to indicate that the product was milled in a manner to eliminate most of the superficial layers and the germ of the grain. "White" bread is the ordinary wheat bread made from "highly milled" flour. "Patent" flour is the word applied by the milling industry to a flour which is of very fine texture and almost white in appearance. It contains practically no bran or germ. By "old-fashioned" Graham flour is meant a wheat flour which is obtained by grinding wheat in buhrstone mills with the omission of bolting. "Whole-wheat" flour is a designation of a flour which contains the whole-wheat berry, with the exception of part of the bran. "First-clear" flour resembles "patent" flour but contains a small amount of the periph- eral portions of the grain. Similar designations have been used for the corn foods. The following tables and charts will illustrate the main results obtained in this investigation- VITAMINE IN CEREAL FOODS. Experiment 1. — "Old-fashioned , ' Graham flour milled in the presence of one of authors ( V) in an old-fashioned buhr mill in Spartanburg, S. C. [Appearance: Typical Graham flour, containing considerable bran and germ. P2O.5 content: 0.866 per cent.] (Beginning with the twenty-flfth day of the experiment, a similar Graham flour was substituted for the above product.) No. of pigeon. First appearance of polyneuritis. Period of observa- tion. Change in body weight in per cent of original weight. Notes. 308 Days. 118 118 123 96 113 + 2 + 9 + 21 + 12 + 13 309 .. .do Do. 312 do 313 do Do. 3il do Do. Remarks. — On the 25th day of the experiment the food was changed to another old- fashioned Graham flour (P 2 O 5 =0.82 per cent), obtained from another buhrstone mill in Spartanburg, S. C. This mill had been in operation for at least 50 years. Slight changes were made in its power plant, substituting turbines for the "water wheels." The process of milling, however, was still the same as at the time of the opening of the mill. Both of these flours contained sufficient antineuritic vitamine to prevent poly- neuritic symptoms and loss of body weight. Their P 2 O s content is almost as high as that of whole wheat. Experiment 2. — " Whole wheat flour" milled in St. Louis, Mo. [Appearance: Grayish white. P 2 5 content: 0.61 per cent.] No. of pigeon. First appearance of polyneuritis. Period of observa- tion. Change in bodyweight in per cent of original weight. Notes. 332 Days. 90 90 91 103 96 +21 -28 -33 -22 -3b 333 do Do. 336 do Do. 337 do Do. 335 do Do. Remarks. — Pigeon 337 died apparently from accidental causes, as the necropsy revealed a piece of copper wire, which had perforated the craw and prevented the animal from eating. The other pigeons appeared in perfect health until a few days previous to end of period of observation. No definite paralytic symptoms were observed in any of these birds and the histological examination of the sciatic nerves also yielded negative results. This flour contained sufficient antineuritic vitamine to prevent polyneuritic symp- toms and fatty degeneration ©f the peripheral nerves. Its P 2 5 content is somewhat lower than that of whole wheat. 3 VITAMINE IN CEREAL FOODS. Experiment 3. — Wheat flour obtained from old-fashioned buhr mill, Roebuck, S. 0. [Appearance: White, with yellowish tinge; contains particles of bran. P 2 5 content: 0.265 per cent.J No. of pigeon. 315 317 31S 319 First appearance of polyneu- ritis. No definite symptoms . Twenty-second day . . . No definite symptoms'. No symptoms Period of observation. Died after 109 days. . . Died after 27 days Killed after 158 days . ....do Change in body weight in per cent of original weight. -3G -15 -28 + 4 Notes. No treatment. Do. Do. Do. Remarks.— Sciatic nerves of pigeons 315 and 319 did not reveal any degenerative changes and were to all appearances normal. Sciatic nerve of pigeon 318 revealed a slight degeneration. This flour represents a fairly "highly milled" product. It was holted through a fine silk cloth, which eliminated a considerable part of the germ and bran. In physical appearance this flour is not as refined as "patent" flour. It obviously contains more antineuritic vitamine than "patent"- flour but not as much as " whole- wheat " flour. The P 2 3 content is also low. Experiment 3a. — Wheat flour obtained from old-fashioned buhr mill, Spartanbura, S. C. Appearance: White with yellowish tinge; contains small particles of bran. P : 5 content: 0.27 par cent.] No. of pigeon. First appearance of polyneu- ritis. Period of observation. Change in body weight in per cent of original weight. Notes. 338 Died after 102 days Killed after 118 days do -13 -12 - 6 - 5 -34 Not treated. 339 Do. 340 Do. 341 do Do. 343 Do. Remarks. — Sciatic nerve of pigeon 339 showed slight degeneration. There were never noticed any definite symptoms of polyneuritis in this bird. Sciatic nerve of pigeons 338 and 340 did not exhibit any degeneration; that of pigeon 341 revealed slight degeneration, although the bird seemed to be in perfect health when it was killed. Experiment 4. — "Patent wheat flour" obtained from a Maryland roller mill. [Appearance: Pure white; no bran can be detected. P 2 6 content: 0.25 per cent.] No. of igeon. First appearance of polyneu- ritis. Period of observation. Change in body weight in per cent of original weight. Notes. 326 Died after 26 days -22 -44 -21 -31 - 7 -20 Relieved symptoms on 327 thirtieth day by an extract of rice pol- ishings. Relieved symptoms on 328 do thirtieth and fifty- seventh days by yeast vitamine. Not treated. 329 Do. 330 do Do. 331 Relieved symptoms on twenty-eighth day by an extract of rice polishings. VITAMINE IN CEEEAL FOODS. Remarks. — Pigeon 326 received, on thirtieth day of experiment, 0.5 cubic centi- meter of Funk's vitamine fraction from an extract of rice polishings. Bird was greatly- improved as a result of this treatment, but gradually developed a relapse several days later. Pigeon 327 received, on thirtieth and fifty-seventh days of experiment, 0.5 cubic centimeter autolyzed yeast vitamine, prepared by means of Lloyd's reagent (dose equivalent to 40 cubic centimeters autolyzed yeast filtrate). The pigeons of this series all developed polyneuritis within three weeks. The flour was "nighly milled" and had a low phosphorus content. The administration of an active preparation of antineuritic vitamine promptly relieved the symptoms. Experiment 5. — "Patent" ivheat flour obtained from a roller mill in Virginia. [Appearance: Pure white; no particles of bran or germ. P 2 O t content: 0.20 per cent.] No. of pigeon. First appearance of polyneuritis. Period of observation. Change in body weight in per cent of original weigh* Notes. 344 Fifteenth day Died after 29 days Died after 30 days Died after 39 days Died after 19 days -37 -34 -43 -37 -14 -28 345 Do. 346 Tenth day Do 347 Do 348 Temporary improve- ment after adminis- tration of "vita- mine." 349 Fifteenth day Remarks. — All of the pigeons of this lot developed typical polyneuritis between the tenth and twenty-second days after feeding was begun, and died with marked loss in body weight. Pigeon 348 was given one dose of 0.3 cubic centimeter of a fluid antineuritic preparation, obtained from autolyzed brewers' yeast, which caused a temporary improvement in the symptoms. Experiment 6. — " Patent flour" milled in a Michigan roller mill. [Appearance: Pure white; contains no bran. P 2 Oo content: 0.20G per cent.] No. of pigeon. First appearance of polyneuritis. Period of observation. Change in body weight in per cent of original weight. Notes. 320 Tenth day Died after 38 days do -27 -13 -43 -38 -30 -43 321 Do. 322 Died after 18 days Died after 40 days Symptoms relieved on the eighteenth day by a preparation made from rice pol- ishings. 323 324 do Do. 325 do twentieth and thirty- third days by a prep- aration made from yeast. Remarks. — Pigeon 322 received 0.5 cubic centimeter of a dilute extract of rice pol- ishings (vitamine fraction), which caused complete disappearance of severe symptoms of paralysis. Pigeon 325 was given, on the twentieth day, 0.5 cubic centimeter of an extract of brewers' yeast (vitamine fraction) ; severe paralytic symptoms disappeared within 24 hours. On the thirty-third day another attack of polyneuritis was com- pletely relieved by a dose of 1 cubic centimeter autolyzed yeast vitamine, prepared 69113°— 18 2 10 VITAMINE IN CEREAL FOODS. by means of Lloyd's reagent (dose equivalent to 40 cubic centimeters autolyzed yeast filtrate). The pigeon remained free from any symptoms after this treatment for about 25 days. Experiment 7. — "Straight patent flour" milled from winter wheat at a roller mill in South Carolina. [Appearance: White; contains a few particles of bran. P2O5 content: 0.216 per cent.] No. of pigeon. First appearance of polyneuritis. Period of observa- tion. Change in body weight in per cent of original weight. Notes. 351 Died after 36 days Died after 38 days Killed after 118 days Died after 52 days Died after 57 days -27 -39 i-34 -28 -48 Not treated. 352 do... Do. 353 Relieved symptoms on 354 • twenty-sixth day by a preparation made from ox liver; on thirty-fourth day with yeast prepara- tion. Diet changed on Fortieth day. Relieved symptoms on twenty-sixth day by a preparation made from ox liver. Not treated. 350 1 Fortieth Day. Remarks. — Diet of pigeon 353 was changed, on fortieth day of experiment, to a wheat flour obtained from an old-fashioned buhrmill in South Carolina. This change caused the pigeon to regain some of the lost weight (original weight 249 grams, weight at period of dietary change 165 grams; weight at end of experiment, 43 days later, 188 grams). Pigeon was killed after 118 days, being free from symptoms of polyneuritis. Sciatic nerve did not reveal any degenerated fibers. This experiment, in conjunction with those preceding, shows that a "highly milled " flour with low phosphorus content leads to polyneuritis within about three weeks. The symptoms are due to a deficiency of this food in antineuritic vitamine, as demonstrated by the curative effect of a small quantity of an antineuritic extract from yeast or ox liver. Patent wheat flour" obtained from a Minnesota roller mill through the open market. Experiment 8 [Appearance: Pure white; no bran can be detected. P 2 Oe, content: 0.25 per cent. [ No. of pigeon. First appearance of polyneuritis. Period of observa- tion. Change in bod j- weight in per cent of original weight. Notes. 1 Days. 25 37 38 38 17 36 -39 -40 -33 -35 -22 -24 Died of polyneuritis; sciatic de- 2 generated. Do. 3 Severe paralysis relieved with 4 antineuritic material. Do. 5 Twelfth day Died of polyneuritis; sciatic de- 6 generated. Do. VITAMINE IN CEREAL FOODS. 11 Remarks. — On this flour the birds developed polyneuritis in about the same time as on highly milled rice. Marked degeneration of the sciatic was found on histological examination. The P 2 5 content of the flour was low (less than one-third that of "whole wheat"). Experiment 9. — "White-wheat bread" obtained from a bakery in Washington, D. C. [Appearance: Good; no bran. P 2 ©5 content: 0.20 per cent.) No. of pigeon. First appearance of polyneuritis. Period of observa- tion. Change in body weight in per cent. of original weight. Notes. 37 Days. 45 80 04 40 35 40 -30 -42 -23 -38 -41 -39 Polyneuritis cured by extract of "whole-wheat bread." Not treated; died of polyneuritis; sciatic degenerated. Severe polyneuritis cured by ex- tract of "whole-wheat bread." Do. 38 39 Fifty-fourth day 40 41 Not treated; died of polyneuritis sciatic degenerated. Do. 42 Remarks. — The bread used in this experiment was made from a mixture of ' ' patent " and "first clear" flour, yeast, sodium chloride, compound lard, molasses, and evapo- rated milk. The exact formulas could not be obtained. This bread had a low phosphorus content and led to polyneuritic symptoms in the pigeons of this seriea in aboutthreeto five weeks. It isseen that the incubation period is somewhat longer (three to five weeks) than in birds kept on "patent" flour (2 to 3 weeks). This incubation period is also longer than that observed when polished rice forms the exclusive diet of pigeons. Three of the pigeons were given an extract of the " whole- wheat " bread used in experiment 15. Chart 3 illustrates the gradual loss in body weight on a diet of white bread and the slow recovery after treatment with the extract of "whole-wheat" bread was begun. The addition of yeast and evaporated milk to the flour, in the process of bread making, did net seem to furnish sufficient antineuritic vitamine to prevent polyneuritis. Experiment 10. — White-wheat bread, ordinary baker's bread, purchased on the open market in Washington, D. C, and Spartanburg, S. C. [Appearance: White; contained no bran. P2O5 content: 0.26 and 0.19 per cent.] No. of pigeon. First appearance of polyneuritis. Died after- Change in body weight in per cent of original weight. 110 120 300 304 305 307 401 402 Thirty-second day. Twentieth day Twenty-fifth day . . Twenty-first day . . Twenty-fourth day Twenty-fifth day . . Fourteenth day . . . Fortieth day Days. Not determined. Do. -15. -29. -4. -18. Not determined. Do. Remarks. — From these experiments it would seem that the incubation period of polyneuritis is somewhat prolonged (about 14 days) as a result of the addition of yeast and evaporated milk to the highly milled flour in the process of bread making. Both yeast and milk are known to contain antineuritic vitamine. 12 VITAMINE IN CEEEAL FOODS. Experiment 11. — IJigh grade "'patent" wheat flour + Lloyd' s reagent (inactive). This flour was obtained from a Minnesota roller mill and 1 per cent of Lloyd's reagent in- corporated with it. [Appearance: White; no coarse particles. P 2 Os content: 0.25 per cent.] No. of pigeon. First appearance of polyneuritis. Period of observa- tion. Change in body weight in per cent of original weight. Notes. 13 Days. 38 38 38 38 38 54 -43 -27 -40 -38 -32 -48 Experiment discontinued after 38 14 days. 15 Died of polyneuritis; sciatic nerve 10 degenerated. Experiment discontinued after 38 17 .do da vs. Do. 18 .do On fifty-fourth day pigeon was dying; killed with chloroform; sciatic degenerated. Remarks. — This experiment shows that "patent" flour, with the addition of Lloyd's reagent (fuller's earth) which has not been activated by autolyzed yeast, does not prevent the appearance of polyneuritis in pigeons. These results are in marked con- trast with experiments illustrated by chart 5. VITAMINE IN CEREAL FOODS. 13 * T -fc f i » 4 J - - a ' - - 4 7 --■'-- 4 i - - V V 5- s _J_ . J_ / 2 ■=1 1 IS4 X S3 i 1 _ Q* V ~\ - - L L - ^ 1 t : ... c t \ 1 - - it I- r 1 13 1- [ 1 - - «"t lizt it t£ i ' . -it •- Zt l v 8 . ... - „ f r t - it £i t 4 ~ 5 ^ V 4 ir^ / / SV' ' 7 ~ L ' Z £ * i A %^ A to •* vN^ ^"=> ^5v *" 1 / if ^\ g\ 7l Q \ - - ^Z LL 1 - - J. 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J^J t/ ^L S \£ i ^-%t ±it-tsr n r\ ^^r-C \ ^^ i ^IS :t Jl-^ / uZ ^ r **_ j j* * -L 1 .' ! » i V- Jl^ & ¥ / IF 't JO. st ft .S U :3 ;SSI v\ ^^ 5^^ -!*_-•' '' f o 5 N 3^' ^*i ^ 4^v ' "Q E U^ ^" 5 %^ "*T M^- * 1 i 1 r - ■■ r% V33yj ~ o •s s ^.g -2 a fcs o "3 J} 69113°- 18 VITAMINE IN CEREAL FOODS. Experiment 17. — Whole corn. [P 2 Oc, content: 0.76 per cent.] No of chicken. First appearance of polyneuritis. Period of observa- tion. Change in body weight in per cent of original weight. 6 Days. 222 428 188 -14 10 do - 5 11 do - 2 Remarks. -^The animals of this group were healthy, full-grown chickens. They were kept in individual small cages throughout the period of observation. The lack of cage space may account for the trivial loss in body weight, as it is very difficult to keep fowls in perfect condition in such narrow confinement for so long a time. Experiment 18. — ' 'Old process water-ground corn meal" obtained from an old-fashioned buhr mill in Georgia. [Appearance: Contains large particles of bran and germ. P-.0 & content: 0.659 per cent.] No. of pigeon. First appearance of polyneuritis. Period of observa- tion. Change in body weight in per cent of original weight. Notes. 66 Days. 150 73 -30 -36 67 do Do. Remarks. — Pigeons 66 and 67 remained perfectly well, but lost considerable weight. This loss of weight was due to the underfeeding of these birds. They received only 10 grams corn meal daily, instead of 15 to 20 grams. In spite of the loss of weight, these animals did not exhibit the slightest signs of polyneuritis during the long period of observation. Experiment 19. — Corn grits (hominy) obtained from an old-fashioned buhrstone mill in South Carolina. [Appearance: Practically all the bran and germ mixed with the finer- endosperm. PsOj content: 0.668 per cent.] No. of pigeon. First appearance of polyneuritis. No symptoms No symptoms; escaped No symptoms Period of observa- tion. Days. 152 98 77 Change in body weight in per cent of original weight. (') Notes. Not treated. Do. Do. 1 Not determined. VITAMINE IN" CEREAL FOODS. 19 Experiment 19a. — Same as used in experiment 19, except that the larger particles of bran were removed by passing through a coarsely meshed sieve. [P2O0 content: 0.59 per cent.] No. of pigeon. First appearance of polyneuritis. Period of observa- tion. Change in body weight in per oent of original weight. Notes. 102 Days. 152 152 152 - 8 - 6 -10 103 do Do. 100 do Do. Experiment 20. — Corn meal used at Milledgeville State Hospital, Milledgeville, Ga. [Appearance: Contains considerable bran and germ. P 2 Oo content: 0.(77 per cent.] No. of pigeon. First appearance of polyneuritis. Period of observa- tion. Change in body weight in per cent of original weight. Notes. 72 Days. 52 84 -38 -15 73 Do. Remarks. — Both pigeons were underfed, receiving only 10 grams of commeal daily. This fact accounts for the loss of body weight. Pigeon 73 also received old process corn meal, obtained from the United Stales Penitentiary, Atlanta, Ga. (0.77 per cent P 2 5 ). This bird remained well and apparently in perfect health for a period of 84 days when the experiment was dis- continued. No symptoms of polyneuritis were exhibited. Experiment 21. — Corn grits obtained from State institutions in Alabama and Georgia. [Appearance: Highly milled. P ; ; , content: 0.169 and 0.201 per cent.] No. of pigeon. First appearance of polyneu- ritis. Period of observation. Change in body weight in per cent of original weight. Notes. 61 1 —22 ( 2 ) -48 -33 62 twenty-second day, daily dose 0.2 c. a yeast extract. Beginning with twen- 64 Died after 31 days tieth day, daily dose 0.2 c. c. yeast extract. Not treated. 411 Do. 1 Twenty-second day of experiment. '■ Not determined. Remarks. — Pigeons 61 and 62 received grits which was used in the State hospital in Alabama (P 2 O s content =0.169 per cent). Pigeon 64 received grits used in a hos- pital for the insane in Georgia (P 2 5 content=0.201 per cent). All of these products contained very little bran and germ and were products of roller mills. Pigeons 61 and 62 responded very well to the treatment with yeast extract (disappearance of polyneuritic symptoms) and were in excellent condition at the end of the period of observation. 20 VITAMINE IN" CEREAL FOODS. Experiment 22. — "Highly milled" corn grits obtained from a roller mill in North Carolina. [Appearance: Contains a trace of bran and germ. P 2 5 content: 0.21 per cent.] No. of pigeon. First appearance of polyneuritis. Period of observation. Change in body weight in per cent of original weight. Notes. 91 Died after 72 days Not determined. . . do Symptoms relieved on 92 Twenty-third day sixty-fourth day by yeast extract. Improved on sixty-sev- enth day by alco- holic extract of yeast. 93 do 94 Forty-seventh day do third day by yeast extract. 97 do on sixty-fifth day by 0.2 gin. alcoholic yeast extract. Experiment 2?>. — Highly milled corn grits and wheat bran. The grits were obtained from a roller mill in North Carolina and the bran from, an old-process {stone) mill in South Carolina. The grits were the same as used in Experiment 22. [P 2 5 content of corn grits: 0.21 per cent. P 2 5 content of wheat bran: 2.48 per cent.] No. of pigeon. First appearance of polyneu- ritis. Change in 1m iily weight Period of observation, in per cent of original weight. Diet. 87 149days do + 3 1 35 gm. corn grits+10 | gm. wheat bran. 85 do 86 ...do... do - 9 88 Died after 68 days (') 226 davs +8 1 40 gm. corn grits+ 5 gm. (' wheat bran. 89 90 One hundred and fiftieth day . . . 1 Not determined. Remarks. — Pigeons 85, 86, and 87 remained in perfect condition throughout the period of observation. Pigeons 88, 89, and 90 developed polyneuritic symptoms after a considerable length, of time and two of the birds died of the disease. Pigeon 89, however, recovered from the paralytic attack, regained its lost body weight and appeared perfectly well at the end of the period of observation (226 days). Experiment 24. — Highly milled corn grits with addition of corn bran. The grits were the same as used in experiment 22, and the bran was obtained from old-process corn meal by passing the latter through a wide-meshed sieve. [P 2 5 content of corn grits: 0.21 per cent. r 2 6 content of corn bran: 0.7S7 per cent.] No. of pigeon. First appearance of polyneuritis. Period of observation. Change in body weight in per cent of original weight. Notes. 81 101 do.. . Died after 113 days Not determined... do Symptoms relieved on seventy-third day by 10 c. c. of dilute yeast extract. Do. VITAMINE IN" CEREAL FOODS. 21 Remarks. — Both pigeons of this lot received a mixture of 15 grams of corn grits and 5 grams of corn bran. The addition of corn bran caused a very considerable delay in the onset of polyneuritis (about 40 days). This furthermore seems to indicate that corn bran is probably not as rich in antineuritic substance as either wheat bran or possibly corn germ. On examination of the corn bran it was noticed that very little of the so-called aleuron layer adhered to the cellulose layer of the bran. Both pigeons recovered temporarily from their paralytic symptoms as a result of a single dose of an antineuritic yeast preparation. The birds again recovered their lost appetite and remained free from symptoms for 38 days (pigeon 81) and 41 days (pigeon 101), respectively. In order to discover whether a larger addition of corn bran would offer complete protection from polyneuritis, pigeon 74 was fed on a mixture of 10 grams highly milled corn grits (0.20 per cent P 2 5 ) and 10 grams corn bran (0.78 per cent P 2 5 ). The corn grits used were the same as fed to pigeon 411 (experiment 21) and had caused symp- toms of polyneuritis on the eighteenth day after feeding was begun. Pigeon 74 re- mained well on this diet for two months, when the experiment was discontinued. Experiment 25. — Corn germ obtained from a roller mill in Indiana. [P 2 5 content: 2.81 per cent.) No. of pigeon. First appearance of polyneuritis. Period of observa- tion. Change in body weight in per cent of original weight. Notes. 70 71 No symptoms do Days. 85 92 -17 -20 Not treated. Do. Remarks. — These pigeons were underfed (see p. 649), receiving only 10 grams of food daily. They never exhibited the slightest symptoms of polyneuritis and were lively throughout the period of observation. Conclusions. — (1) The results obtained in this investigation clearly show that for pigeons an exclusive diet of whole wheat or corn fur- nishes an adequate supply of antineuritic vitamine. (2) The antineuritic vitamine seems to reside in the peripheral layers and the germ of these seeds, whereas the endosperm is relatively poor in this substance. (3) If wheat and corn foods containing only a small percentage of the peripheral layers and germ of the seed are fed to pigeons and chickens exclusive of other food, polyneuritic symptoms appear on an average of three weeks after the beginning of the feeding period. The appearance of polyneuritis is preceded by a gradual loss in body weight. The birds can be relieved of their paralysis in a striking way by the oral or subcutaneous administration of a highly concen- trated preparation of antineuritic vitamine derived from "whole- wheat" bread, yeast, ox liver, rice polishings, or beans. (4) The addition of yeast (in amounts used by bakers) in the preparation of bread from highly milled flour does not prevent the appearance of polyneuritis in birds fed on this food, but prolongs slightly the period of incubation. 22 VITAMINE IN CEREAL FOODS. (5) The addition to "highly milled" flour, or bread made from "highly milled" flour, of a small amount of antineuritic vitamine preparation will correct this particular dietary deficiency, and will prevent the appearance of polyneuritis and the loss of body weight. (6) The total phosphorus content of corn and wheat foods is a fairly satisfactory index of the amount of antineuritic vitamine con- tained in these foods. In a general way, it can be said that a high total phosphorus content is an indication that the particular corn or wheat product is relatively rich in antineuritic vitamine References. Holst, 1907, J. Hyg., vol. 7, p. 629. Little, 1912, J. Am. M. Ass., vol. 58, p. 2029. 1914, ibid., vol. 63, p. 1287. Ohler, 1914, J. Med. Research, vol. 31, p. 239. Simpson and Edie, 1911-12, Ann. Trop. Med. & Parasit., vol. 5, p. 321. Seidell, 1914, Public Health Reports, February 18, p. 364. Voegtlin, Sullivan, and Mters, 1916, Public Health Reports, August 14, p. 935. Wellmann and Bass, 1913, Am. J. Trop. Dis. & Pre v. Med. THE GROWTH-PROMOTING PROPERTIES OF FOODS DERIVED FROM CORN AND WHEAT. 1 By Cakl Voegtlin and C. N. Myebs, Division of Pharmacology, Hygienic Laboratory, United States Public Health Service. The authors in a preceding paper have shown that the whole wheat or corn grain contains an abundance of antineuritic vitamine, whereas the "highly milled" products derived from these cereals are deficient in this respect. This conclusion was drawn from experi- ments on adult animals. As growing animals require the presence of all dietary constituents to a greater extent than do adults, it seemed desirable to continue the investigation along this line. Recent studies on growth have furnished the necessary information to answer the question as to what constitutes a diet complete enough to insure growth. It is now generally held that a physiologically suffi- cient diet must contain an adequate caloric value derived from protein of proper composition, carbohydrate, and fat. In addition it must contain a sufficient amount of antineuritic and fat-soluble vitamines and of necessary inorganic salts. The dietary deficiencies of the whole wheat and corn kernel in the diet of growing animals have already been investigated. Thus Hart and McCollum (1914), working with young albino rats and hogs, have shown that normal growth is obtained when the wheat kernel is supplemented by the addition of inorganic salts, fat-soluble vita- mine, and casein. Subnormal growth was observed with rations consisting of wheat plus casein and salts; wheat plus casein and butter fat; wheat plus salts and butter fat. McCollum, Simmonds, and Pitz (1916) have made similar observa- tions with corn. In this case also the grain requires an improvement in its protein moiety, its salt content, and an added supply of fat- soluble vitamine. Of these three additions the correction of the deficiency in certain inorganic salts seemed to be of the greatest importance, inasmuch as this correction in itself furnished a ration on which rats did grow fairly well for several months. Hart, Halpin, and Steenbock (1917) report experiments with pullets weighing 2 to 3 pounds on corn and wheat rations. These investigators found that corn and wheat meal do not support growth in this species of animals; fortifying the cereals with an appropriate i Reprint from the Public Health Reports, vol. 35, No. 22, May 31, 1918, pp. 843-868. 23 24 VITAMHSTE IIST CEEEAL FOODS. inorganic salt mixture, protein of proper composition, and a small amount of fat-soluble vitamine (2 per cent butter fat) leads to normal growth. These authors also call attention to the possibility of intoxication as a result of excessive wheat feeding, which they attribute to the presence of some toxic substance in the wheat kernel. The purpose of the present investigation was to answer the ques- tion as to whether the corn and wheat products used in human nutri- tion exhibit similar dietary deficiencies as those of the whole grains. The bulk of the corn and wheat foods of the American dietary are derived from the wheat and corn kernel by means of a process of milling (roller mills) which is known to eliminate most of the germ and superficial layers of the grain. It, therefore, seemed to us a question of practical importance to determine whether the milling process improves, or causes a decrease in, the dietary value of the milled product. Moreover, it was desirable to decide whether or not the food additions made to flour (yeast, salt, milk) in the preparation of bread improve the nutritive value of this food. Experimental. The experiments were carried out on squabs, young albino mice, and a few hogs. Most of the work was done with wheat, and only a few incomplete experiments were made with foods obtained from corn. We are not aware of any previous records where squabs have been used for studies on growth. For this reason the following details are given, as they may be of interest to workers in this field. The growth period of pigeons is extremely short, as will be seen from the records. Almost maximum body weight is reached, on an adequate diet, within 40 days after the squabs are hatched. The feathers develop gradually, and by the time full growth is reached the body is completely covered. The birds begin to fly at the age of about 40 days. This is usually followed by a slight loss of body weight, which is probably due to the strenuous muscular work performed in flying. It should be pointed out that squabs can not feed themselves during the first two weeks of life. During this period the parents feed the young by regurgitating food which has been softened in their crops. The function of the crop consists in the preparation of the cereal food for gastric digestion. It is very doubtful whether active digestion takes place within the crop, and it is more likely that this organ simply softens the food by means of water in order to facilitate its disintegration by the stomach. In the absence of the crop, gastric digestion of cereals would be a rather slow process, even in the case of the bird's stomach with its powerful muscular wall and the gravel which takes the place of millstones. In order to obtain a sufficient number of squabs for experiments on growth, about 70 to 100 healthy pigeons were kept in a well-ventilated room containing numerous cages (wire screened), the doors of which were left open. Usually the birds built their nests in these cages and after laying the eggs the parents were caught, confined to the cage, and put on the diet to be tested for its growth-promoting properties. When it happened that eggs were laid outside of the cages, the pigeons were allowed to hatch and the young squabs were then transferred, with the nest and the parent birds, to the nearest cage. We found that it was impossible to move the eggs, as the birds refused to sit after the eggs had been handled. The sitting period of pigeons is about 17 days. The male bird sits from about 9 a.m. to 5 p.m.; the female from 5 p. m. to 9 a. m. VITAMINE IN CEREAL FOODS. 25 The stock pigeons were fed for nearly two years, while this work was in progress, on an exclusive diet of corn and wheat, crushed oyster shells, and river sand. Fresh running water was supplied. On this diet the birds bred very well throughout the year, with tho exception of the moulting season. The average egg weighed about 16 grams ; the contents, minus the shell, weighed about 15 grams, and the weight of the squab immediately after hatching was about 13 grams. It was found that squabs which were below the average body weight and those that had difficulty in hatching, were of low vitality. The squabs were weighed as soon after hatching as possible and every three days thereafter during the morning hours (9 a. m.). The growth curve during the first 10 days is almost a straight line, slight deviations being due to variations of crop content. The birds which died as a result of a diet of inadequate composition were necropsied and the sciatic nerves examined for the presence of myelin degeneration. The great advantage of using squabs for growth experiments is due to the possibility of immediately starting the newborn animal on a ration which is to be investigated. It is obvious that this can not be done when mammals are used. The experiments with albino mice were carried out on young animals weighing approximately 6 to 8 grams and obtained from a healthy stock kept in the labora- tory. Recent work has sufficiently demonstrated that mice are as well suited for growth experiments as albino rats. The animals were placed in wide glass jars with a wire screened top. Sawdust was used as bedding. Under these conditions the mice could be kept in excellent health on an adequate diet for nearl- a "ear. The animals were weighed every three or four days. 1 The hogs used were young animals, either purchased on the open market or raised in the laboratory. They were kept in stalls with a concrete floor. The bedding was wheat straw, some of which was eaten by the animals. Tap water was supplied. The experiments on hogs are somewhat complicated, as the straw and tap water may perhaps be considered as a source of inorganic salts and fat-soluble vitamine. How- ever, this factor remained approximately constant in each experiment. Food used.— When the whole wheat was fed to mice it was usually crushed in an ordinary kitchen mill. The wheat flour, either alone or with other foods, was made into cakes by means of water. The wet cakes were dried at 45° C. and broken up into small pieces. The white flour used was bought under the name of "patent" flour and came from one of the largest roller mills of the West. The wheat "mid- dlings" were obtained from a roller mill in Washington, D. C. The "whole wheat" bread used was purchased in Washington, D. C, and the bakery volunteered the following information in regard to the food materials used in the preparation of this bread: Standard loaves were made from crushed whole wheat, with the addition of canned eggs, some salt, olive oil, molasses, compound lard, wheat bran, and pressed yeast. The accurate proportions of the various constitutents could not be obtained. The "white" bread was made from "highly milled" wheat flour, with the addition of sodium chloride, compound lard, yeast, and evaporated milk. For 900 standard loaves of this bread, 588 pounds of flour, 47 pounds of evaporated milk, and 8| pounds of pressed yeast were used. In the experiments where the "white" bread was supplemented by other foods, the bread was fust dried at 40 to 50° C, crushed in a mill, and mixed with the other food in the desired proportions. The casein was a purified preparation made in this laboratory. The crushed oyster shells, fed to the pigeons, consisted largely of calcium carbonate, with traces of organic matter. When the rations contained chemically pure calcium carbonate in place of the oyster shells, exactly the same results were obtained so far as the growth of squabs was concerned. The grit was i The normal growth curve of mice was taken from the article by Mitchell, J., Biol. Chem., 191G, vol. 26, p. 24. 69113°— 18 4 26 VITAMINE IN CEREAL FOODS. well-washed river sand, consisting mainly of silicates. The so-called "activated" Lloyd's reagent was used as a source of antineuritic vitamine. 1 In some experiments the "inactive" Lloyd's reagent was included in the rations. Fuller's earth, which had not been in contact with the yeast filtrate, was used in this case. The highest proportion of activated Lloyd's reagent contained in any of the rations was 3 per cent. In most cases 0.6 per cent or 1.5 per cent were used with equal success. When the activated Lloyd's reagent forms 0.6 or 1.5 per cent of the ration, the total nitrogen derived from this source represents only 15 to 37 milligrams. It should be pointed out that squabs do not well tolerate the addition of consider- able quantities of fat to the diet. In some experiments, where 5 per cent of lard was incorporated in the ration, the birds showed diarrhea and poor growth, followed by decline and death. Summary. It seems superfluous to enter into a detailed discussion of the results obtained in this investigation, as the accompanying charts illustrate the results in a comprehensive manner. For this reason only the main points will be referred to in this summary. 1. The "highly milled" products are, without exception, inferior in dietary value, as regards growth, to foods prepared from the whole grain. It is rather surprising that such delicate organs as the gastro- intestinal tract of young mice can tolerate a diet containing a large amount of bran. This fact, however, does not necessarily mean that it is advantageous to include the bran in foods intended for human nutrition. On the contrary, the experiences with "war bread" would rather indicate that persons with delicate digestion are sub- ject to temporary digestive disturbances as a result of a change from "white" bread to bread containing a considerable percentage of bran ("war bread"). On the other hand, from the standpoint of dietary completeness, a bread including all of the grain, with the exception of the superficial cellulose layer, is undoubtedly superior to the so-called white bread, made from "highly milled" flour, and would not possess the above-mentioned objectionable features. 2. The "white" bread used in these experiments was not adequate for maintaining normal growth, in spite of the fact that it was pre- pared with some evaporated milk and yeast. The most significant defect of "white" flour is the deficiency in antineuritic and fat- soluble vitamine; it is also deficient in adequate protein and inor- ganic salts. • 3. A wheat flour, containing a considerable part of the germ and superficial layers of the grain, supports growth of mice and pigeons especially well when supplemented with inorganic salts. Tho same is true of "whole wheat" bread. i It was prepared from autolyzcd brewers yeast by treatment with hydrochloric acid and nitration. This yeast nitrate was then treated with a special grade of fuller's earth, which removed a considerable part of the active vitamine from the yeast filtrate. The dried preparation was free of protein and gave negative tests for tryptophan, cystin, and tyrosin. No lysin could be isolated but the activated Llloyd's reagent contained a substance which in its reactions resembled histidine. The total nitrogen oontent of the dried reagent was about 2.5 per cent, and this consisted largely of adenine and other basic substances derived from yeast filtrate. VTTAMINE IN CEREAL FOODS. 27 4. "Highly milled" corn grits, forming the exclusive food of young hogs, leads to failure of growth in these animals, whereas the whole corn kernel, supplemented by inorganic salts, promotes growth. 5. Newborn squabs are suitable animals for growth experiments. 6. No evidence of a toxic action of a whole wheat diet was obtained in the experiments on squabs which were fed on whole wheat meal, supplemented by a suitable salt mixture. 7. In the light of our present knowledge, it would appear that bread made from "whole wheat" flour, or old-fashioned corn meal, should be used in preference to "white" bread and "highly milled" corn foods, whenever the diet is restricted to these cereal foods to the more or less complete exclusion of other foods possessing greater dietary values. Bibliography. Hart. Halpin. and Rteenbock. 1917. J. Biol. Chem., vol. 31, p. 415. Hart and McCollum. 1914. J. Biol. Chem., vol. 19, p. 373. McCollum, Simmonds, and Pitz. 1916. J. Biol. Chem., vol. 28, p. 153, Mitchell. 1916. J. Biol. Chem., vol. 26, p. 24. 28 VITAMINE IN CEREAL FOODS. U. 5. Public Health ££fiv/cc Chart 1.— Shows satisfactory growth of mice when "whole wheat " bread is supplemented With 5 per cent of salt mixture. Evidently the Whole wheat bread used in this experiment Was slightly deficient in inorganic salts, as seen from chart 2, where the bread was fed without the addition of the salt mixture. The composition of the salt mixture used in this investigation was as follows: NaCl, 0.50 gm.; KjjHPC^, 1.21 gm.; CaH 4 (PoO, H 2 0, 0.256 gm.; Calcium lactate, 2.944 gm.; Ferric citrate, 0.100 gm. Chart 2.— Illustrates the growth of mice on "whole wheat" bread. The retardation of growth is mainly due to the deficiency of this diet in inorganic salts. (See chart 1.) VITAMINE IN CEREAL FOODS. 29 30 VITAMINE IN CEREAL FOODS. Chaht 4.— Illustrates the subnormal growth of mice on a diet of "under milled" flour. This flour was obtained from a roller mill and was bought as "second clear." It contained 0.E2 per cent P 2 & . From the phosphorous content of this flour it would appear that this product is the grade of flour intermediate between a "first and second clear." Chart 5.— Shows insignificant growth of mice when "white" flour is supplemented with a salt mixture. The orange juice was added to the drinking water with the idea of preventing scorbutic symptoms. The "white" flour was bought under the name of "patent" flour and contained 0.25 per cent of P..0 5 . The sciatic nerves of mouse 102, C6, and £7 showed marked myelin degeneration. These animals probably died of polyneuritis. VITAMINE IN CEREAL FOODS. 31 Chart 6.— Illustrates early failure of growth, followed by death of mice on a diet of "white" flour plus Salt mixture The sciatic nerves of these mice showed marked myelin degeneration. The results of this experiment are therefore identical with the one which is illustrated by chart 5. Chart 7.— Growth of mice. Illustrates beneficial effect of the addition of antineuritic vitamine in the form of activated Lloyd's reagent to a mixture of "white" flour and inorganic salts. Compare with charts 5 and 6. Bread No. 24 is not a complete diet, probably deficient in fat-soluble vitamino and certain essential amino acids. 32 VITAMINE IN CEREAL FOODS. Chart 8. — Practically no growth (except in mouse 149) was obtained on a diet of ''white" bread and salt mixture. The addition of 5 per cent butter fat (as indicated by arrows) caused considerable growth in mouse 150. Mice 145 and 149 showed somewhat better growth as a result of the butter-fat addition, but both animals died later on in the experiment. The sciatic nerve of one animal of this group and which was examined after death showed considerable myelin degeneration. (Indication that this diet is deficient in antineuritic substance.) Chart 9.— Shows beneficial effect on growth of mice as a result of the addition of casein and lard to bread No. 27 (see chart S). All of the animals died, however, with the exception of mouse 174. The " white" bread is evidently deficient in antineuritic vitamine and amino acids, essential for normal growth. Bread No. 2 is also deficient in antineuritic vitamino (compare with chart 11). Bread No. 2 is same as bread 27, except that the former is supplemented by casein and lard. VITAMINE IN CEREAL FOODS. 33 Chart 10.— Illustrates deficiency of bread No. 1 in antineuritic vitamine. Compare with chart 12. All the mice of this series died after periods ranging from 16 to 93 days. Chart 11.— Bread No. 7, which is deficient in fat-soluble vitamine, leads to fairly good growth in mice. Mice 196 and 201 showed normal growth for a considerable length of lime (120 days) and reached nearly maximum weight. The other mice of this series stopped growing after having been on this diet for 40 days. 34 VITAMINE IN CEREAL FOODS. Chart 12.— Illustrates the fairly good growth of mice on bread No. 8. Completion of growth was not obtained with all the mice of this series. Mice 204 and 206 did not grow well after having reached about 12 gm. oi body weigh'. Chart 13.— Growth of mico. VITAMINE IN CEREAL FOODS. 35 Chart 14. — Shows retardation of growth of mice on bread No. 10, deficient in inorganic salts. 36 VITAMINE IN CEREAL FOODS. Smjs. 38o 36o 34-0 320 3oo 2 So z6o Z4o zzo Zoo /So /6o /4o 120 loo fa (0 4o to Ch *RT 15. ^S Dm CON 7 W4 "S >- ,t/j A f/ ' f i tf\ / T 1 k\ r ■--y, Discc NT'N. JEO- - ^ t * L> ^ ^ IP y f i l f /J ! 4 J / J {/ 1 \ 1 / / \ \ 1 | ' / \o e/>o. l i i 1 1 ' ?' J 1 f i | 1 I / / HH ■.AT SO ' i / Co ?/V. SO J / Or 0,1 5T£i* '/r. S/i£ '-LS. i i | I* 1 - V M<> Chart 15. — Shows satisfactory growth of squabs on a diet limited to the corn and wheat kernel, supple, mented by the addition of calcium carbonate (oyster shells). The oyster shells were crushed and fed to the parents ad libitum. Squab B22 reached normal body weight in 20 days, but died suddenly at the age of 75 days. The cause of death is unknown. The other two squabs of this series showed normal growth and development and lived for 150 days, when the experiment was discontinued. The appearance of the birds at this time was normal in every respect. VTTAMINE IN" CEREAL FOODS. 37 f hart 16.-Shows failure of growth of squabs on a diet of whole wheat alone. The birds evidently suffered from privation of mineral salts. The skeleton was poorly developed, calcification being very deficient. When the wheat kernel was supplemented by the addition of calcium phosphate practically normal growth was obtained. Sciatic nerve of Squab Bll, B5, and B6 shows no myelin degeneration. S3 VITAMINE IN CEREAL FOODS. 3So 36o 34o 3Z0 3oo 28o 2 60 240 220 Zoo /So /bo Mo /20 /ao So 60 !4o 20 . Ck \RT • 7 nam i ^ P 7 t V / w >^ "»/ CJ 1 $7 1 1 r I / / / 7 \ / / \ / i 1 | A 1 r / 1 1 / / < / Hho i*/tf tf/«r ftWW. 3. j / \ / / a 1 * # Chart 17.— Shows delayed growth and failure of growth of squabs on "whole wheat bread." This food is deficient in calcium salts. Compare with chart IS. Sciatic nerve of squab A15 did not reveal any myelin degeneration. VITAMINE IN CEKEAL FOODS. 39 Sms. 3to 36o ■34-0 520 3oo 180 Z6o 2-4 o Ch \HT 18. ^ /O&CA . A s ^r / \ A s /■ / %-<■ V V aV X "h \ / * V ,-■*" —7 ••s^ ..-' ...»• v .~4_ § p J> ( ^-' .-■•- ^ V I i 1 1 > ?/ i • i i - ^' - 9t13. 320 *£* - / < % « - V •lOQ j / : ^ lio /(So 1 - :- 1 i • I . / --• <* /So loo So 6o 40 zo O / / I ... / Tvta LE t <|V£* -'/9* '.no- J i « ?.S/»A ire. J / j U* Chart IS.— Illustrates normal growth and development of squabs on a diet of "whole wheat" bread and calcium phosphate. The parents of these two squabs raised two other pairs of squabs while being fed on this diet. This proves that growth, reproduction, and maintenance of normal nutrition are possible on a simple diet as "whole wheat" bread supplemented by calcium salts. 40 YTTAMHSTE IN" CEEEAL FOODS. SflS. 38o d bo Ch. \RT n WOAYS "^ \ 30O Z8o Z6o 14o 110 Zoo /8o /bo 140 IZO 100 So bo 40 ?o y t / f V- $ /v* V \ / f V ^._ / i $ *} II i\'HI r £ F . /*■/■ 'MTC0 IS ( I I A* v P fi f Chart 20.— Illustrates failure of growth, followed by death of squabs, on a diet of " white " bread and cal- cium phosphate (A33, A37, and A3S). When the "white" bread is further supplemented by casein and a preparation containing antineuritic vitamine, normal growth and development results. Squabs B70, 72, 811, 833 reached nearly maximum normal weight at the age of about 30 days. Sciatic nerves of A33, A37, and ASS exhibit myelin degeneration. 42 VITAMINE IN" CEREAL FOODS. 38e 36o SAO 32X> «3oo Z2o Z60 240 220 Zoo l2o J60 140 IZo loo So 60 40 2.0 Ch ART 21. iODm j \ l/ \/ / \ \ / V *~r^ v "-— ♦ t f ^ I \ y\, A $ r n S2 !>"'— # ^ it r 1 f 1 1 it / \ 1 / f 1 / / \ I | 1 \ [ \ • / I 1 \ 1 1 | \ 1 1 j f \ 1 /V«/7 1. IAO" )?2 1 1 / \ I r//v ■ 'Mix TURL 1 ! | i 'Llo\ /l£/>C ■oil/ em) CTIW reo 30 1 / / f .* r A*' ^ Chart 21.— Shows satisfactory growth of squabs when the "white" bread is supplemented by protein of proper composition, inorganic salts, and antineuriticvitamine. In this experiment the amount of tha latter food accessory is twice as large as in the experiments illustrated by chart 20. VITAMINE IN CEREAL POODS. 43 Sms. 39o 06o 040 3Z0 3oo 28o I60 140 220 2 00 /So /So 140 /ZO loo So 60 4o 2o Ch M*T 22. » S5£! t f •"«■. ~°5*i — ~. h * >■••■*, j.y \ r 1 1 y v- r u fl 1 1 / A V r* 1 > 1 ■■ / / ! f 1 / / 1 / / t / 1 r ■ / / 1 / / / / \ 1 I 1 j j \ i / 'h/nr re" a iEAD 7^-5 / / 15.0 S.c SALtMi ~rom / / PCACt Iff :r/vA *a I.S 1 1 1 f 1 ^ 1 v£ # ^ Chart 22.— Growth of squabs. The diet used in this experiment mug be considered as physiologically complete. The "white" bread evidently contains sufficient fat-soluble vitamine (derived from evapo- rated milk) to render this ration adequate for growth. 44 VITAMINE IN" CEREAL FOODS. Sns. 3So 3&0 340 320 300 28o 26o 140 zzo Zoo I80 /bo 140 IZo 100 So 60 Zo Cha \T 23. -\ 1 1 ,oa •w } 1 \ 1 f f\ \ J~ V if 1 1 \-* *v / - 1 fl / r i / [[ 1 i I 1 1 1 • ,' / HHITt "bke to — t> o *> j s r> i i I I* 3 — — * i .* & II \ K ,$ ft K "*** ~ — * •^^ ^"**« ""■— — ■—__._ < o &vs 'W*2 "t553 3^T" =Q (3 ft • 0°, ■s .an fe ft § 2 a ° S a 'S pot* a o « lift o ": g lid 7 *3 c3 a ; i s J f8 *> O S ^ >o < ' sf ~TPi> 'VOfy/ ll§Iall§ll c 1 § i o o o o o CO -O * M P O CD | sl II 3 ! ~ n 2 « tj o Q => F 85 a do g &-; S « S •S o, § -J s ft .13 1 3 u, b «i w « B -I- 3 rn ft 2 a ." o (I ■° U Q,'-3 K a> a ° I "3 o a S js ft I « c if -2 VITAMHSTE IN CEREAL FOODS. 47 Chart 29 : We CHT C URVf a , p G ] ii" 40 U 3o in to OS) r-s too 200 |i $ ?» ? f I' a — *? § ; J k q 5 5* 2 ~i 1 ■« 15 5 I V $* q ' / ■-• \ r * * — 1 ' ' /' N -J- A t 1 / s~ \ /"» s >i V 1 s "'•^ 1 / / I \ ""S « r "1 e L, i J - f ( ' f©W r w ,,. ) * > c ? t / / 2 § ;- g / *J~ 5 6 / ? 9 ; i / 5 ■■■z fOa W « ?■< « •^ Chart 29.— Illustrates the failure of growth of young hogs on a diet of "highly milled'' corn grits. This food was obtained from a roller mill and represents the endosperm of the corn kernel. The animal was born of a hog which had been raised in the laboratory on a diet of cracked corn, wheat straw, and tap water. On the corn grits the animal did not gain more than a few pounds during nine months, in spite of the addition of a supplementary salt mixture and antineuriticvitamine. The animal finally died. The necropsy revealed the following abnormalities: Emphysema of lungs, chronic gastritis, small injected areas in small intestine, chronic passive congestion of liver, congestion of spleen, no scorbutic changes. Sciatic shows marked myelin degeneration. Another pig of the same litter showed a similar growth eurve on a diet of corn grits. Here also correction of the salt content and antineuritic vitamine of tho diet did not prevent death. Necropsy findings were the same as in Fig IVB. 48 VITAMINE IN CEREAL FOODS. Chart 30 FOLLOWED : -Weight Curve: of by Whole Corn. pigh oh Diet of Corn OilCake, LBS. 190 180 170 ibO 150 140 130 (2o no too go 80 70 60 60 40 30 20 10 o Days 100 200 300 400 <500 bOO 700 80O 900 . *1 1 1 1 vl B 5 J t 1 * 1 cs < uat " f> X C> I