we pointed out that the nutritional anemia induced in rabbits by a whole milk PDF
Preview we pointed out that the nutritional anemia induced in rabbits by a whole milk
IRON IN NUTRITION. 1V. NUTRITIONAL ANEMIA ON WHOLE MILK DIETS AND ITS CORRECTION WITH THE ASH OF CERTAIN PLANT AND ANlMAL TISSUES OR WITH SOLUBLE IRON SALTS.* BYE. B. HART, C. A. ELVEHJEM, J. WADDELL, AND R. C. HERRIN. (From the Department of Agricultural Chemistry, University of Wisconsin, Madison.) (Received for publication, December 21, 1926.) In an earlier publication (1) we pointed out that the nutritional anemia induced in rabbits by a whole milk (cow) diet could not be corrected by the addition of inorganic iron (Fez03) but that such an anemia could be corrected when in addition to the Fez03 there was added to the diet either fresh cabbage or a cold 95 per cent alco- holic extract of desiccated cabbage or an alcoholic extract of corn- meal. These extracts were practically free from iron. A chloro- phyll preparation free from iron fed in the presence of added ferric oxide was also an effective supplement to a whole milk diet as a prophylactic for anemia. From these experiments we deduced the conclusion that some organic nucleus ‘necessary for hemoglobin building was absent or deficient in whole milk. This deduction was in harmony with the generally accepted theory (but for which there was no experimen- tal proof) that there may be a deficiency in whole milk of some nucleus constructed from a multiple of pyrrole groups and which is necessary for hemoglobin building. It was this idea which led us to add a preparation of chlorophyll to whole milk supplemented with FenOa and which gave us positive results in respect to anemia prevention. As our work on this problem progressed, we had accumulated dat,a showing that not only were the corn grain and fresh cabbage * Published with the permission of the Director of the Wisconsin Agri- cultural Experiment station. 299 This is an Open Access article under the CC BY license. 300 Iron in Nutrition. IV effective agents in the prevention of anemia in the rabbit con- fined to a whole milk (cow) Fez03 diet, but that lettuce was also especially effective as a preventive of anemia. We had included lettuce in our category of supplements because Evans (2) and as- sociates had used it with marked success as a source of vitamin E and we were not sure at that time but that there might be some direct relation between vitamin E and our problem of milk defi- ciency and nutritional anemia. TABLE I. Potency of Fresh Lettuce in the Cure of Nutritional Anemia. - Rt-zTit HISlO- Data Diet. globin. I Zrythrocytes Weight. - -- -- -- 198.5 per e.mm. om. 94 June 12 Whole milk only. 365 “ 25 8.9 4,500,000 640 July 1 5.9 3,550,ooo 780 “ 2 Added Fe203 (0.015 gm. daily) + 50 gm. fresh lettuce daily. “ 10 8.5 6,300,OOO 990 “ 16 9.8 6,275,OOO 1170 “ 17 Discontinued; in perfect condition. - - - We tried to feed dried lettuce in order to demonstrate that drying would not injure its potency, but the dried material seemed to be very unpalatable and the rabbits would not eat the preparation. In Table I is shown a typical record of the cure of an anemia induced in the rabbit by a whole milk-Fe203 diet, by the use of fresh lettuce. The amount of hemoglobin and the erythrocyte count increased almost twofold in a period of 2 weeks. Before making further attempts to separate the active principle of these plant materials functioning as cures or preventives of anemia on a whole milk-Fe203 diet, it was decided to determine whether the ash of these materials would be effective. We had tentatively assumed that the preventive principle was of an organic character. Our work up to the present has been mainly with lettuce ash although the ash of other materials has also been tried. Hart, Elvehjem, Waddell, and Herrin 301 So successful have been the cures or preventions of nutritional anemia induced by a whole milk-Fe203 diet with the ash of certain materials, that we feel certain that the deficiency of cow’s milk in respect to its anemia-producing properties is not an organic principle but something of inorganic character resident in the ash of certain plant and animal tissues. These ashes of plant and ani- mal materials were prepared in an electric mufRe operating at a temperature of 650-750°C. and for a period of 20 to 30 minutes and were devoid of carbonaceous materials. On occasions the ashes were prepared in gas-heated furnaces at temperatures of a cherry- red. The use of such temperatures must make it certain that we are dealing in these experiments with inorganic materials and not with organic substances. EXPERIMENTAL. For all the experiments to be described in this paper rabbits have been used. We have also used chicks confined to a special diet as checks on the work with the rabbit, but the ration used and the results secured with chicks will be described in a future publication. The young rabbits were always weaned from the doe at 4 weeks of age and placed on the experimental diet. The stock ration fed the does during the period of gestation consisted of 100 parts of rolled oats, 10 parts of wheat bran, 1 part of common salt, and whole milk ad libitum. Green materials (cabbage, green grasses, or carrots) were given the mother until the young rabbits were about 3 weeks old, but were withheld during the last week of the suckling period. The experimental animals were confined individually in cages fitted with $ inch mesh screen bottoms to prevent feces consumption, and were weighed regu- larly throughout the experiment. They usually weighed 300 to 400 gm. when placed on the experiment. In some cases we have periodically irradiated the experimental animal with ultra-violet light for the purpose of protecting against rickets. We have found that this irradiation had no influence in protecting the animal against an anemia on a whole milk diet. The practice of irradiating our experimental animals has not been general because on the diet used, there has been no direct evidence that our animals suffered from rickens. Sodium citrate, used in our earlier experiments to Iron in Nutrition. IV prevent the formation in the stomach of large curds and possible gastritis, has not been used in our later experiments. We have found it unnecessary. Our experimental colony has suffered some from intestinal diarrheas terminating in death, but the records presented in this paper are from animals free from such disturb- ances. Hemoglobin was estimated by the Fleischl-Miescher hemoglobinometer and expressed as gm. per 100 cc. of blood. A discussion setting forth the desirability of expressing the hemo- globin content of blood by a uniform standard will be found in a paper by Elvehjem and Waddell (3). Erythrocytes were deter- mined in an American Standard hemocytometer with Levy count- ing chamber. The young rabbits 4 weeks old were given whole milk ad libitum until the gm. of hemoglobin per 100 cc. of blood had decreased to 4.7 to 5.8 and the erythrocyte count to 3.5 to 4.5 millions. This generally took a period of from 5 to 6 weeks. At this time the milk was supplemented daily with 0.015 gm. of Fe20a and the lettuce ash. The ash from 5 gm. of ground dried leaf lettuce was triturated with the Fez03 in a small quantity of milk and was fed each morning. This insured a more complete consumption of the supplement. The animals were fed individually. During the re- mainder of the day the rabbits were allowed an excess of milk. Table II contains the records of two of the rabbits and is typical of the improvement noted after feeding the lettuce ash to anemic rabbits. Rabbit 180 after showing definite anemic symptoms was fed lettuce ash plus FezO$ directly. Rabbit 164 upon show- ing similar symptoms was fed Fe203 alone for a period of 19 days to again show that Fez03 fed as such was ineffective in improving the blood stream. Then the lettuce ash was added to the diet. One may readily see from the table that as soon as the ash of lettuce was added there was a steady improvement in the blood stream and after a period of approximately 30 days the amount .of hemo- globin and erythrocyte count was normal. After the blood stream became normal, it remained so, provided the ash of lettuce was fed. R.abbit 180 was maintained in perfect condition for 118 days and Rabbit 165 for a period of 111 days. Although discontinued at 1 The Fez03 used was the same material used in all of our work and was a Baker and Adamson product marked C.P. Hart, Elvehjem, Waddell, and Herrin 303 TABLE II. Potency 0f . Lettuce Ash in the Cure of Nutritional Anemia. Date. Diet. gHlo.Cb?inK. UJ- Erytbrocytes Weight. 1966 per c.mm. gm. act. 5 Whole milk only. 425 “ 16 7.2 4,600,OOO 540 Lt 22 5.9 4,050,000 660 “ 23 Added Fez08 (0.015 gm. daily) + ash of 5 gm. dried lettuce. “ 29 6.3 3,950,ooo 725 Nov. 6 7.4 4,820,OOO 770 “ 12 9.3 4,900,000 870 “ 24 9.8 5,150,000 1150 Dec. 9 10 .o 5,970,ooo 1200 “ 29 12.3 6,650,OOO 1065 19.26 Jan. 6 11.0 6,170,OOO 1300 “ 13 11.5 6,700,OOO 1385 “ 20 11.7 6,200,OOO 1510 “ 29 11.5 6,350,OOO 1560 Feb. 10 11 .o 6,250,OOO 1790 “ 18 11.9 6,450,OOO 1850 19% Sept. 29 Whole milk only. 590 Oct. 9 8.1 5,250,OOO 660 “ 20 7.0 5,300,000 840 ‘I 26 7.6 5,200,OOO 850 Nov. 5 7.6 5,100,000 860 “ 8 6.3 3,700,000 860 “ 18 Added Fez03 (0.015 gm. daily). I1 27 5.1 3,100,000 975 Dec. 2 5.1 2,770,OOO 1020 “ 7 5.1 2,920,ooo 1020 ‘I 8 Added Fe203 (0.015 gm. daily) + ash of 5 gm. dried lettuce. “ 14 7.2 3,520,OOO 1040 (( 22 7.6 4,000,000 1160 “ 29 7.8 4,200,OOO 1225 304 Iron in Nutrition. IV TABLE II-conduded. - - - RFFt Date. HgleoRbiIno.- 1X rythrocytes Weight. _- - - -_ g?n. per 1996 100 cc. per c.mm. 8”. blood 164 Jan. 6 8.3 4,600,OOO 1335 “ 13 9.1 5,150,000 1460 “ 20 11.1 6,070,OOO 1470 “ 29 10.4 5,800,000 1625 Feb. 10 10.4 5,700,000 1900 “ 18 10.2 5,800,000 2060 Mar. I 8.5 5,500,000 2250 “ 17 9.8 5,900,000 2310 ‘( 29 10.2 6,000,OOO 2450 that time there is no reason to believe that the normal condition of their blood stream would not have been maintained indefinitely. Although similar results were obtained with several other rab- bits, the rapid improvement of the blood stream and the continued maintenance of their good health produced merely by the addi- tion of the inorganic elements in the lettuce ash with the Fe&& was almost unbelievable. We were slow in being convinced that this was true since our earlier results and those of several other investigators had led to the conclusion that the factor limiting hemoglobin building on a milk diet must be of organic nature. Therefore we immediately started further experiments in order to substantiate our data. More animals were used and greater pre- cautions taken to minimize the role of possible genetic factors entering into the experiment. An entire litter of young rabbits at weaning time was placed directly on the diet plus the various supplements. The preventive type of experiment was used. The litter consisted of eight animals. Each animal was given daily the whole milk plus 0.015 gm. of Fez03. Two rabbits were given no further addition, two were given the ash of 5 gm. of dried lettuce, two the ash of 5 gm. of dried cabbage, and two the ash of 5 gm. of yellow corn. This gave us an opportunity to compare rabbits from the same lit.ter, all of which had received the same preexperimental feeding. The ex- periment also made possible a comparison of the potency of the ash of lettuce, cabbage, and corn on an equal basis. Two such Hart, Elvehjem, Waddell, and Herrin 305 series were started at different times and the animals in each ex- periment represented a single litter. The results of the entire experiment were exceedingly uniform. The data obtained from the two rabbits in each group and the two groups in each series were so comparable that we present only the data from one rab- bit in each group. These results are shown in Table III. Again it was found that the ash of lettuce was exceedingly beneficial in the prevention of nutritional anemia and the main- tenance of a normal blood stream in a rabbit on a whole milk- Fe203 diet. Rabbit 301 on the basal diet survived only 148 days, during which period the average amount of hemoglobin was 8.0 FIG. 1. Rabbit 302, reared on a ration of whole milk-FepOa plus the daily administration of the ash of 5 gm. of dried lettuce. Appears to be a perfect specimen. Hemoglobin normal and erythrocytes per c.mm. about 5,700,OOO. Started on the ration at 450 gm., and after 9 months on this ration weighed 2400 gm . and the erythrocyte count 3,810,OOO. Rabbit 302 receiving in addition to its whole milk-Fe203 diet the ash of 5 gm. of dried lettuce made good growth and maintained a normal blood stream over 200 days or 7 months and at this writing is continuing its growth. The average gm. of hemoglobin per 100 cc. of blood dur- ing the period was 10.4 and the erythrocyte count 5,950,OOO. The rabbit appears normal in every respect. Fig. 1 is a photo- graph of this animal taken September 13,1926. The results seem to us exceedingly remarkable. This animal received no roughage, TABLE III. - lhtc. Iht. Erythrocytcs. Weight. yw per lHZ6 100 cc. ym. blood 301 1”cl). 17 \Vholc milk + Vc,Os 530 (0.015 gm. daily). “ “7 s.7 4,700,000 GGO I\lar. 10 7.0 3,000,000 980 “ l!) 8.7 4,100,000 1090 “ 26 x5 4,600,OOO 1190 :\I”‘. 5 7.G 3,850,OOO 910 “ 16 7.2 3,iOO,OOO 860 ‘I “-1‘( 5 5 'L,GOO,OOO 950 Alay 3 72 3,800,000 1100 “ 12 4,s 2,400,000 1200 “ “4 i.G 4,000,000 1360 JUllC s 9 .3 4,800,000 1540 (( L’, s.5 4,300,000 li50 July 5 7.4 4,~00,000 1)520 “ 15 G.8 3,000,000 1750 July ‘24 Dead. ‘I’ypical record on \vhole rnilk-l”c,08-lettuce ash diet. 302 Pcl,. 17 \Yhole milk + FelOy 4i0 (0.015 gm. daily) + ash of 5 gm. dried lettuce. (6 "7 10.0 6,200,OOO G80 hlar. 10 0.3 5,400,000 1000 “ 19 S.9 5,600,OOO 11'70 “ 36 10.6 6,200,OOO 1320 i\Pl'. 6 10.0 6,400,OOO 1420 “ 15 10.4 5,800,000 1460 “ 23 10.8 6,500,OOO 1560 May 3 12.1 7,200,OOO 1650 “ 12 12.1 7,400,000 1670 “ 3I 4 10 2 5,700,000 1730 June 8 10.0 5,600,OOO 1800 (I 21 10.4 5,700,000 2020 July 5 10.4 5,400,000 2200 “ 15 10 2 5,900,000 2310 “ 30 s.9 5,500,000 2280 Aug. 17 10.0 5,700,000 2450 “ 30 10 2 5,600,000 2510 Sept. 14 10.4 5,400,000 2520 Oct. 4 0.3 5,500,000 2520 “ 19 98 5,500,000 2400 Nov. 3 10.2 5,700,000 2390 306 Hart, Elvehjem, Waddell, and Herrin 307 TABLE III-Concluded. RF2it Date. Diet. Erythrocytes. Weight. Typical record on whole milk-FeeOs-cabbal ash diet. gm. per 1966 100 cc. per c.mm. blood 305 Feb. 17 Whole milk + Fez03 470 (0.015 gm. daily) + ash of 5 gm. dried cabbage. “ 27 8.3 4,400,000 640 Mar. 10 8.9 4,700,000 840 ‘( 19 9.6 5,800,OOO 990 “ 26 8.3 5,000,000 1170 Apr. 5 10.2 5,700,000 1300 “ 15 10.6 6,000,OOO 1460 “ 23 10.6 6,200,OOO 1770 May 3 9.1 5,800,OOO 1900 “ 12 9.6 5,600,OOO 1900 (‘ 24 11.1 5,800,OOO 1880 June 8 10.0 5,600,OOO 1980 “ 21 9.3 5,400,000 2070 July 5 9.3 5400,000 2170 “ 15 9.8 5,800,OOO 2370 “ 30 10.0 6,000,OOO 2400 Aug. 17 9.8 5,500,000 2370 “ 30 8.5 4,500,000 2430 Sept. 14 10.0 5,600,OOO 2440 Typical record on whole milk-Fe203-corn grain ash diet. 295 Jan. 22 Whole milk + Fe203 (0.015 gm. daily) + ash of 5 gm. corn. Feb. 3 8.5 5,825,OOO 570 “ 10 8.9 6,000,OOO 600 “ 18 8.1 4,700,000 590 ” 27 6.6 4,500,000 640 Mar. 9 6.8 4,600,OOO 810 “ 26 6.3 3,200,OOO 830 Apr. 3 6.1 3,100,000 830 “ 13 6.6 3,500,000 930 May 3 7.8 4,200,OOO 960 “ 18 7.0 3,800,OOO 1050 May 26 Dead. 308 Iron in Nutrition. IV having been kept on a wire screen, and yet from all outward ap- pearances, such as sleekness of coat, appetite, etc., appeared as a normal specimen. Reproduction has not as yet been studied. The ash of 5 gm. of cabbage was nearly as effective as that of lettuce. This is shown in the record of Rabbit 305 (Table III). Although the amount of hemoglobin and erythrocyte count was TABLE IV. Typical Record on Whole Milk-Fe203 + Ash of10 Gm. of Dried Cabbage Diet. Date. Diet. gIIloebmino.- Zrythrocytos Weight. gm. per f9M 100 cc. per c.mna. om. blood 314 Mar. 9 Whole milk + FezOx 580 (0.015 gm. daily) + ash of 10 gm. dried cabbage. “ 20 7.2 4,300,000 660 “ 29 11.1 6,500,OOO 860 Apr. 15 10.6 6,000,OOO 980 “ 23 11.3 7,000,000 1180 May 3 10.6 6,500,OOO 1400 “ 12 9.6 5,900,000 1610 “ 24 8.7 4,700,000 1700 June 8 10.9 6,000,OOO 1950 “ 21 10.6 6,400,OOO 2090 July 5 10.4 6,100,OOO 2120 “ 15 10.0 5,700,000 2230 “ 30 10.2 6,100,OOO 2170 Aug. 17 10.6 5,800,OOO 2270 Sept. 14 9.3 5,100,000 2370 Oct. 4 9.1 5,000,000 2450 “ 19 10.6 5,900,000 2340 Nov. 3 9.8 5,700,000 2290 - slightly lower for this animal than for Rabbit 302, it is still in our colony, continues to grow, and appears to be normal. Table IV shows the results secured with Rabbit 314 which was one of the individuals in the later series. This rabbit received the ash of 10 instead of 5 gm. of dried cabbage. It was found that, on this higher level of feeding the amount of hemoglobin and the
Description:The list of books you might like
