TX 556 .115 J48 ; Copy 1 TREASURY DEPARTMENT UNITED STATES PUBLIC HEALTH SERVICE HUGH S. GUMMING, Surgeon General ANTINEURITIC VIT AMINE IN SKIM MILK POWDER BY J. M. JOHNSON Chemist AND C. W. HOOPER Pathologic Physiologist Division of Pharmacology, Hygienic Laboratory United States Public Health Service REPRINT No. 689 FROM THE PUBLIC HEALTH REPORTS August 26, 1921 (Pages 2037-2043) 7 -'^ L WASHINGTON GOVERNMENT PRINTING OFFICE 1921 A"* ANTINEURITIC VITAMINE IN SKIM MILK POWDER.' By J. M. Johnson, Chemist, and C. W. Hooper, Pathologic Physiologist, Division of Pharmacology, Hygienic Laboratory, United States Public Health Service. Reconstructed milk has recently come into use in localities where an ample suppl}^ of fresh milk is not available. This milk is made from skim milk powder, butter fat, and water, mixed in the propor- tions necessary to re-form liquid milk. The mixing is carried out in a specially designed machine, so that a separation of the butter fat takes place only upon long standing. It became necessary to decide whether skim milk powder used in making reconstructed mil'c retains its water-soluble vitamine (B) content or whether th;s is destroyed by the process of drying. We have, therefore, attempttM to determine the amount of skim milk powder necessary to prevent polyneuritis in pigeons. Vedder and Clark ^ found that 5 c. c. daily of fresh cow's milk or 5 c. c. of canned milk did not prevent polyneuritis in fowls. E. A. Cooper ^ fed fresh milk to pigeons and found that 35 c. c. daily only delayed the onset of polyneuritis until the fiftieth day. Protection was not secured by this amount. Gibson and Concepcion ^ found that the minimum protective amount of liquid cow's milk for fowls is between 100 and 200 c. c. daily. They concluded that milk con- tains such a small amount of antineuritic substance that care should be used to extend the diet of infants as early as possi])le. They did not find any advantage of raw milk over autoclaved milk in this respect. EXPERIMENTAL. Skim milk powder made by the spray process was used. The pol- ished rice was picked over by hand and all incompletely polished grains were carefully removed. The rice was then ground in a small mill and mixed in varying proportions with the skim milk powder. A small amount of water was added to make a sticky dough. This was dried out under the electric fan and broken into small pieces. The pigeons ate this food very readily. The table shows the results obtained from the feeding of rice and various percentages of skim milk powder. All of the 6 control pigeons fed on 100 per cent rice developed symptoms of polyneuritis and 1 Reprint from the Public Health Reports, vol. 36, No. 34, Aug. 26, 1921, pp. 2037-2043. ' Philippine Jour. Sci. VII, B, 423 (1912). •Jour. Hyg. XIV, 12(1914). 'I'hilippine Jour. Sci. XI, B, 119 (1916). 2 67777°— 21 LIS«ARY OP CONGi^ESS DOCUMENT* UiViilON ANTINEURITIC VITAMINE IN SKIM MILK POWDER. 3 died within 48 days. Histological examination of their sciatic nerves showed the presence of fatty degeneration. Of 7 pigeons fed on a diet composed of 80 per cent rice and 20 per cent skim milk powder, 5 showed fatty degeneration of the sciatic nerve and 1 showed a normal nerve; the seventh bird escaped, but had shown no symp- toms of polyneuritis after 139 days. The onset of polyneuritis in 1 case of the 5 cases was delayed until the one hundred and twenty-eighth day. Of 4 pigeons fed on a diet composed of 75 per cent rice and 25 per cent skim milk powder, 3 showed fatty degeneration of the sciatic nerve and 1 a normal nerve. However, the onset of polyneuritis w:as delayed in the 3 cases from 57 to 82 days. Of 16 pigeons fed on diets CmHTl :- (00^ RicE. composed of 30-100 per cent skim milk powder and 70-0 per cent rice all showed normal sciatic nerves. However, in 3 cases, symptoms of polyneuritis with subsequent recovery were observed. An examination of the table will show that in order to secure com- plete protection from polyneuritis the skim milk powder must consti- tute at least 30 per cent of the diet. A diet containing 25 per cent skim milk powder will delay the development of polyneuritis but will not protect against it. Even 20 per cent skim milk powder delayed the onset in a few cases. The pigeons consumed about 20-25 grams daily of the mixture of. skim milk powder and rice. Therefore it requires about 6-7 grams daily of skim milk powder to protect from polyneuritis. This corresponds to about 75 c. c. of liquid milk. ANTINEURITIC VITAMINE IN SKIM MILK POWDER. ANTTNEURITIC VITAMINE IIST SKIM MILK POWDER. \ \ 1 • 1 • < 1 \ • 1 / • / 1 Jd t Y 1 / / / • \ ; • 1 < Q o -1 f r 1 • / • / « % i; • 1 • / • / i i / • / r o \ / • • 1 < z • 1 1 t o o o 1 «-:t/wtoi7-* ANTIlSrEURITIC VITAMINE IN SKIM MILK POWDER. Results of experimental feeding of pigeojis^ Food. Symptoms of polyneuritis. •Length of life. Termi- nation. Pigeon No. Per cent rice. Per cent skim milk powder. Sciatic nerve. 32 100 100 100 100 100 100 80 SO SO 80 80 80 80 75 75 75 75 70 70 70 70 60 00 60 50 50 50 40 40 40 After 16 days After 31 days After 32 days After 24 days After 14 days After 39 days After 128 days.... After 27 days After 24 days Emaciated by tenth day. Days. 24 37 48 31 16 41 130 46 28 12 7 139 84 187 99 65 84 102 102 102 102 102 102 101 101 101 101 102 101 102 S3 83 44 Died.... ...do ...do ...do ...do ...do ...do ...do ...do ...do ...do Escaped Died.... Killed... Died.... ...do..... ...do Killed... ...do ...do ...do ...do ...do ...do ...do ...do ...do ...do ...do ...do ...do ...do Died .... Extensive fatty degeneration (.Marehi). Do. 33 34 Do 38 Do. 43 Do. 40 Definite fatty degeneration (Marehi) Do. Moderate fatty degeneration (Marehi). Do. Definite fatty degeneration (Marehi). 42 35 36 37 39 20 20 20 20 20 20 20 25 25 25 25 30 30 30 30 40 40 40 50 50 50 60 60 60 100 100 100 41 do 47 44 45 46 48 19 After 81 days After 121 days; re- covery. Emaciated after 60 days. After 57 days After 82 days None Definite fatty degeneration (Marehi). Normal. Moderate fatty degeneration (Marehi). Definite fatty degeneration (Marehi). Extensive fatty degeneration (Marehi). Normal. 20 do Do. 21 . ..do Do. 22 23 After 32 days; re- covery. None Do. Do. 24 do Do. 25 do Do. 26 do , Do. 27 do.. . Do. 28 29 After 32 days; re- covery. Do. Do. 30 do Do. 31 do Do. 10 do Do. 11 ..do Do. 12 Questionable at 42 days. Do. Since Cooper found that 35 c. c. of fresh milk delayed the develop- ment of polyneuritis only until the fiftieth day, and Gibson and CH/j«r4 :- 70% Rice - 5oy» Milk Powder. ¥^ N^ ^^ \^- A ^ ~^^ o '<^' 'V 1.0 t>fiVS-> \ •-»-. N._c^ / Concepcion found that fowls required 100-200 c. c. of fresh milk daily for protection, it appears that even fresh milk contains very little antineuritic substance. Our work shows that the amount of ANTINEURITTC VITAMINE IN SKIM MILK POWDER. 7 this substance in skim milk powder is of the same order of magni- tude as in fresh milk, and that therefore the process of drying skim milk does not lead to an appreciable destruction of the vitamine in question. ' CHARTS:- 60% Rice - +0% Skim Milk Powder. i .1 " ^^ \ V v.--- .' — \ v .'^y V... ^--^y ^*«— — •^•- ^^ .^ /-^ \z .r ♦- 2o Days- :;^ „,•-•— s^ 'n,^«-«»v«^ .^P^ Chart 6:- 5o% Rice-5o7« 3kim Milk Powder. ^#26 *2V^.. W28 •\. -.>*v ./V v.^. N. ./ \ v ,^ V / ^.i?' ,•—• ^.^ ChartT:- 407o Rice - 6o7o Milk Powder. »30.., ^\. ^7^ \ N^^ V o W3/ v.^'-^/N/V^-N / ^-"^, 20 cays \ The feeding of rats on a purified basal ration plus milk, by Osborne and Mendel ^ and one of the present authors," shows that a large amount of milk is required to give nonnal growth to albino rats. If '•Jour. Biol. Chem. XXXIV, 537 (1918). «J-. M. Johiisou, unpublished work. 014 487 243 5 ANTINEURITIC VITAMINE IN SKIM MI we accept that the antineuritic and water-sok same, we would expect to find that it'requhea milk to prevent polyneuritis in pigeons, an assumption which is in accord with the facts presented in this paper. The pigeons weighed about 300 grams each. With a requirement of 6 to 7 grams daily for the pigeons, a child of 4.5 k. body weight would require 1,125 c. c. daily of reconstructed milk in ordpr to receive an adec{uate amount of antineuritic substance, provided that the require- ments of the two species were the same. The charts give the picture of the weight changes of the pigeons. The curves are plotted upon the basis of per cent, starting with the initial weight as 100 per cent. CHARTS:- »oo % Skim Milk Powder. •**, If 10 V \x *l/\ .y'^.- =\ V ../•--'V ..-/\ /\_..-' \^.^.,: o \ Mil 2o Ocys — ^ ^» ■X.--' /■\ SUMMARY. Pigeons fed upon mixtures of spray process skim milk powder with polished rice require 30 per cent of the food in skim milk powder in order to get full protection from polyneuritis. This corresponds to about 75 c. c. daily liquid milk. ACKNOWLEDGMENTS. The thanks of the authors are due to Dr. Carl Voegtlin, Chief of the Division of Pharmacology, under whose directions this work was carried out and who aided in many ways during the progress of the work. o LIBRARY OF CONGRESS e 014 487 243 5