UNIVERSITY OF CALIFORNIA 
 
 COLLEGE OF AGRICULTURE 
 
 AGRICULTURAL EXPERIMENT STATION 
 
 BERKELEY, CALIFORNIA 
 
 VITAMIN-A DEFICIENCY 
 
 AS RELATED TO REPRODUCTION 
 
 IN RANGE CATTLE 
 
 G. H. HART and H. R. GUILBERT 
 
 BULLETIN 560 
 
 OCTOBER, 1933 
 
 UNIVERSITY OF CALIFORNIA 
 BERKELEY, CALIFORNIA 
 
Digitized by the Internet Archive 
 
 in 2012 with funding from 
 
 University of California, Davis Libraries 
 
 http://www.archive.org/details/vitaminadeficien560hart 
 
VITAMIN-A DEFICIENCY AS RELATED TO 
 REPRODUCTION IN RANGE CATTLE 2 
 
 G. H. HAET 3 and H. E. GUILBERT 4 
 
 INTRODUCTION 
 
 For a long time after the discovery of vitamins and the subsequent ad- 
 vance in the knowledge of mineral metabolism, the livestock industry 
 tended to regard these findings as largely technical matter with little or 
 no practical application. Evidence in recent years has shown, however, 
 that certain conditions of very great economic importance, spontaneous- 
 ly appearing in livestock, can be explained only by a knowledge of the 
 role of vitamins and minerals in maintaining the well-being of domestic 
 animals. The energy and protein values of f eedstuffs, based on digestible 
 nutrients, are an entirely satisfactory means of evaluating feeds when 
 the rations happen to contain the additional essentials of an adequate 
 diet in sufficient amounts, and account for the importance placed on 
 variety by the husbandman. Unfortunately, however, these additional 
 feed essentials are liable not to be present in adequate amounts in the 
 semiarid southwestern parts of the United States during the dry season 
 when animals are kept entirely on the natural vegetation of the range. 
 
 Experiments with small laboratory animals show that the delicate 
 physiological phenomena of reproduction may be more or less seriously 
 affected by general undernutrition or by specific deficiencies. The repro- 
 ductive failure may vary in character with the nature and degree of the 
 deficiency, and there are four general types of its manifestations : 
 
 1. Failure of the females to come into estrum. 
 
 2. More or less regular occurrence of estrum and mating, with failure 
 in fertilization. 
 
 3. Mating and fertilization followed by death of the fetus and abortion 
 or resorption at various stages of gestation. 
 
 4. Failure in lactation. 
 
 Reproductive failure in cattle induced by lack of phosphorus has been 
 clearly demonstrated in different parts of the world. The evidence on 
 
 i Eeceived for publication May 16, 1933. 
 
 2 The experimental work reported in this paper became cooperative with the United 
 States Department of Agriculture Bureau of Animal Industry, July 1, 1929. 
 
 3 Professor of Animal Husbandry and Animal Husbandman in the Experiment 
 Station. 
 
 * Assistant Animal Husbandman in the Experiment Station. 
 
 [3] 
 
4 University of California — Experiment Station 
 
 phosphorus and protein deficiencies and on general undernutrition as 
 related to reproduction under range conditions in California has been 
 discussed by the authors in previous publications/ 1 ' 2) 
 
 This bulletin summarizes information on the nature and occurrence 
 of vitamin A, the experimental evidence on the relation of vitamin A to 
 reproduction in cattle, data on the symptoms and differential diagnosis 
 of vitamin-A deficiency, and evidence that this deficiency contributes to 
 reproductive failure in range cattle. 
 
 NATURE AND OCCURRENCE OF VITAMIN A 
 
 In 1914 McCollum and Davis (3) found that vitamin A was contained 
 in the unsaponifiable fraction of milk fat — that is, the portion that does 
 not form soaps when heated with alcoholic potash. This evidence that 
 the essential substance was not a fatty acid glyceride but something dis- 
 solved in the fat formed the basis of the terminology, "fat-soluble vita- 
 min A." 
 
 Subsequent investigations by several workers demonstrated that the 
 vitamin could be extracted from plant sources by fat solvents, such as 
 ether and alcohol. 
 
 The occurrence of the vitamin associated with yellow pigmentation, 
 especially in yellow corn, and its absence in white corn, led to research 
 on the possible relation between the vitamin and these yellow pigments. 
 As early as 1826 Wachenroder (4) had extracted and described the yel- 
 low-orange pigment of carrots. In 1864 Stokes (5) discovered a method of 
 separating the yellow pigments of plants from the accompanying green 
 chlorophyll, and in 1866 Piccalo and Lieben (6) extracted and crystallized 
 the yellow pigment from corpora lutea (yellow body of the ovary) of 
 cows. 
 
 In 1911 Tswett (7) proposed the name "carotenoids" for all pigments 
 related chemically to carotene, the pigment of carrots. Escher (8) in 1913 
 demonstrated that the pigment of the corpora lutea of cows' ovaries was 
 identical with carotene from carrots. 
 
 During the years 1914 to 1920 Palmer (9) and coworkers found caro- 
 tene to be the principal pigment in the blood serum, body fat, and milk 
 fat of cattle, and also in the skin secretion of Guernsey cattle. In chickens, 
 on the other hand, the principal pigment of skin, shanks, body fat, and 
 egg yolk proved to be xanthophyll, a closely related substance. Goats, 
 swine, and sheep have little or none of either pigment, while human adi- 
 pose tissue and blood serum contain both pigments in amounts tending 
 to vary with the quantity of each in the diet. Palmer further established 
 that the source of these pigments in normally pigmented species was the 
 
Bul. 560] Vitamin- A Deficiency in Range Cattle 5 
 
 feed. Chickens of breeds which are normally pigmented were raised de- 
 void of yellow color, and they produced eggs devoid of yellow yolk color 
 on carotenoid-free rations. Mead and Regan, (10) experimenting with 
 Jersey and Holstein cattle, have produced body and milk fat without 
 yellow pigment on a ration consisting solely of concentrates. 
 
 Steenbock and his associates, early impressed with the relation of vita- 
 min-A potency to yellow pigment in plants, published a series of articles 
 showing the close correlation between carotenoid pigment and vitamin 
 A in roots, (11) maize, (12) leaves, (13) and peas, (14) as determined by feed- 
 ing pigmented and colorless varieties to albino rats. About the same 
 time, however, Drummond (15) failed to improve the condition of rats 
 suffering from vitamin-A deficiency by feeding pure crystalline caro- 
 tene. Later experiments by Steenbock, Sell, and Buell (16) with milk fats 
 of high and low color, practically colorless cod-liver oil, and light-colored 
 egg yolk failed to show any relation between vitamin-A potency and the 
 amount of pigment present. 
 
 Meanwhile, workers were following a new line of investigation that 
 has led to the present advance in knowledge. In 1920 Rosenheim and 
 Drummond (17) in England stated that the well-known color reaction of 
 sulfuric acid with cod-liver oil applied also to liver fats of other animals, 
 and suggested a relation of the color-producing factor to vitamin A. 
 Drummond and Watson, (18) after three years, were convinced that the 
 relation was more than a casual one but, because the color produced was 
 transient, could not obtain quantitative results. In 1925 Drummond, 
 Rosenheim, and Coward (19) found that arsenic chloride gave, with cod- 
 liver oil, a more stable blue color, the intensity of which indicated a rela- 
 tion to the vitamin-A content. Later that year Rosenheim and Drum- 
 mond (20) gave the first definite technique for a quantitative chemical 
 assay of vitamin A in oils. In 1926 Carr and Price (21) substituted anti- 
 mony trichloride for the arsenic chloride and otherwise improved the 
 technique for estimating vitamin A. 
 
 These discoveries, presenting a simple, rapid method of assaying oils 
 for their vitamin-A content, in contrast to the tedious, expensive, time- 
 consuming feeding tests with animals, stimulated widespread and inten- 
 sive research concerning its validity. Spectrographic studies were made 
 on solutions containing the vitamin and carotenoids, and there was re- 
 newed interest in the relation of the vitamin to the carotenoid pigments. 
 
 In 1928 Euler, Euler, and Hellstrom (22) reported that feeding of 
 carotene to vitamin-A deficient rats restored growth. Their findings 
 were confirmed by Moore (23) and others in 1929. Duliere, Morton, and 
 Drummond, (24) however, using a technique similar to that employed in 
 Drummond's 1919 experiments, failed to induce improvement in vita- 
 
6 University of California — Experiment Station 
 
 min-A deficient rats by carotene feeding". Their failure was subse- 
 quently shown to result from the employment of a solvent (ethyl oleate) 
 in which carotene rapidly deteriorates. 
 
 In 1930 Moore (2rj) fed rats on a vitamin- A deficient diet until they 
 showed signs of the deficiency; at autopsy the livers of a number of 
 them were shown to be devoid of vitamin A by means of the color test 
 with antimony trichloride. The diet of the remaining- animals was sup- 
 plemented with purified carotene. Deficiency symptoms disappeared, 
 growth was resumed, and the livers of these animals were subsequently 
 shown to be rich in vitamin A. Thus it was demonstrated that the pig- 
 ment carotene is converted within the animal to vitamin A, which is 
 practically colorless. 
 
 It is now clear that such species as swine, sheep, goats, and rats al- 
 most completely convert these carotenoid pigments to colorless sub- 
 stances and store in their bodies or secrete in their milk the practically 
 colorless vitamin A. Cattle, on the other hand, convert part of the caro- 
 tene in the feed to vitamin A and store part unchanged in the liver and 
 adipose tissue or secrete it in the milk. Breed differences in this regard 
 are typified by the highly pigmented milk fat and body fat of Jersey 
 and Guernsey cattle as compared with that of Holstein cattle under the 
 same feeding conditions. The biological value of the milk fat is there- 
 fore not necessarily correlated with the degree of color. 
 
 Whatever the origin of the widespread prejudice against yellow fat 
 in the beef trade, it does not appear justified in light of present knowl- 
 edge of nutrition. Aside from hereditary variations, the degree of pig- 
 mentation is governed largely by the quantity of carotenoids ingested 
 with the feed and by a tendency to accumulate pigment with advancing 
 age. Since carotene, the yellow pigment in beef fat, is a source of vita- 
 min A in the human diet, discrimination against it appears inconsistent. 
 The common appearance of yellow color associated with grass-fed beef 
 lacking in finish and with dairy cattle has led to the use of color as an 
 index of quality. Cattle fed heavily on grains containing little or none 
 of this pigment, will eat relatively small amounts of forage normally 
 rich in pigment, thus producing carcasses with light-colored fat. That 
 color is not necessarily associated with low quality is shown by the fact 
 that animals raised at the University Farm and awarded high honors at 
 the Chicago International Livestock exposition were found on slaughter 
 to have yellow fat. The grading of beef based upon conformation of the 
 carcass, quality, smoothness, and degree of finish or covering with fat is 
 sufficient to place all carcasses, including excessively colored carcasses 
 of Jersey and Guernsey cows, in their proper place without regard to 
 the color of the fat. This fat color may be the one point of superiority of 
 
Bul. 5G0] Vitamin- A Deficiency in Range Cattle 7 
 
 the carcasses of these breeds from the standpoint of human nutrition, 
 though the concentration of the vitamin or its precursors even in highly 
 colored fat is relatively low as compared with that in liver or green 
 plant tissue. 
 
 Occurrence of Vitamin A or Its Precursors. — Vitamin A or its pre- 
 cursors are widely distributed in nature. Carotene is present in all 
 green plant tissues, in carrots, and in other yellow roots. Seeds in gen- 
 eral contain little vitamin A. Yellow corn contains considerable amounts, 
 but wdiite corn contains very little, while barley, wheat, rye, oats, and 
 sorghum grains contain only small amounts, entirely inadequate for the 
 nutritional requirements of animals. In animal tissues vitamin A is 
 much more highly concentrated in the internal organs and glands than 
 in the lean tissue or body fat. Liver tissues of most animals are so rich in 
 it that the liver has come to be looked upon as its principal storehouse. 
 The liver oil of cattle which have been fed rations rich in vitamin A has 
 a much higher concentration than does cod-liver oil. 
 
 Conditions Which Destroy Vitamin A or Its Precursors. — Vitamin A 
 and carotene are easily oxidized in the presence of air and light, and the 
 rate of oxidation is accelerated by heat. The curing of hay under condi- 
 tions of exposure to sunlight, which involves bleaching of the green 
 chlorophyll, rapidly diminishes its vitamin-A potency/ 20 ' Coward (27) 
 found that vitamin A is completely destroyed when the leaves of plants 
 dry up, become brown, and die. It is relatively stable to heat in the ab- 
 sence of oxygen. (28) According to Dann (29) it is resistant to oxidation in 
 ethyl alcohol, alcoholic potash solutions, and ethyl acetate; and its po- 
 tency is not diminished by aeration at 98° Centigrade for one hour in 
 alcoholic potash solution. In this connection the influence of antioxygens 
 in stabilizing vitamin A must not be overlooked. 
 
 Because of wide variations occurring in naturally cured hays accord- 
 ing to the degree of exposure, sweeping generalities regarding the differ- 
 ence in vitamin-A potency of field-cured and artificially dried hays 
 should not be made on the basis of experiments involving a few samples. 
 Likewise the widely quoted statement of Jones, Eckles, and Palmer (30) 
 that straw is a good source of vitamin A for cattle requires qualifica- 
 tion. In the midwestern and eastern United States grains are cut and 
 bound or headed as soon as possible after seed maturity. The plants may 
 not be completely dead when harvested, some stems are partly green, 
 and other green plants may be included with the straw. In California, 
 grains ripen after the close of the rainy season, little or no green growth 
 of any kind is present in some areas when the grain is ripe, the plants 
 die, and the standing grain may be exposed to intense heat and sunlight 
 
8 University of California — Experiment Station 
 
 dor two to eight weeks before harvesting. The straw consequently may be 
 very deficient in vitamin A. 
 
 Corpus Luteum and Carotene. — The presence of carotene in the cor- 
 pus luteum is of historical interest, since this was the first animal tissue 
 from which the pigment was extracted in crystalline form by Piccalo 
 and Lieben. (G) Since the corpus luteum is so intimately associated with 
 the rhythmic functioning of the ovaries and with pregnancy, one may 
 logically inquire into the possible function of carotene in this body. 
 Moore (31) recently stated, in connection with an investigation on the 
 presence of carotene and vitamin A in corpora lutea of cows, "the pres- 
 ence of such considerable amounts of carotene in the corpus luteum, 
 apparently unaccompanied by vitamin A, must present an interesting 
 problem in the mechanism of reproduction." Definite proof that caro- 
 tene is the corpus-luteum pigment is available only for cattle and sheep. 
 Bergh, Muller, and Broekmeyer, (C2) and Palmer (9) found no caroten- 
 oids at all in the corpora lutea of swine — an argument against any spe- 
 cific reproductive function. We have examined an ovary carrying the 
 corpus luteum of pregnancy from an animal in the experiment of Mead 
 and Regan (10) on rations devoid of roughage for dairy cattle. The body 
 and milk fat of these animals, as previously stated, were devoid of yellow 
 pigment. Neither carotene nor vitamin A could be demonstrated in the 
 corpus luteum from this particular cow. Others fed similarly have com- 
 pleted normal reproduction. Cod-liver oil was practically the sole source 
 of vitamin A in these rations. Carotene would therefore appear to be 
 present in the corpora lutea of cattle rather because of an affinity for 
 the fatty substances in these bodies than because of a specific function 
 in the mechanism of reproduction. 
 
 REVIEW OF LITERATURE ON RELATION OF VITAMIN A TO 
 REPRODUCTION IN CATTLE 
 
 Definite disease syndromes have been described as caused by lack of 
 vitamin A in certain species of farm animals. Thus Beach (33) in 1924 
 demonstrated that nutritional roup in chickens results from this defi- 
 ciency. In areas where yellow corn is not available, its incidence is rather 
 high unless a green feed is regularly supplied. Hughes, Aubel, and Lien- 
 hardt (34) in 1928 described typical symptoms in hogs which failed to get 
 sufficient vitamin A in the diet. Their elaborate experiments covered a 
 period of seven years. 
 
 Evidence regarding the necessity of this vitamin in reproduction in 
 mammals has been definitely established, but the findings become par- 
 ticularly confused in the case of the bovine species. 
 
Bul. 560 j Vitamin- A Deficiency in Range Cattle 9 
 
 Hart, McCollum, Steenbock, and Humphrey (35) in 1911 published 
 their original work on the physiological effect on growth and reproduc- 
 tion in cattle of well-balanced rations from the wheat, corn, and oat 
 plants respectively, and from mixtures of the three. Reproduction was 
 most seriously interfered with in the animals on the wheat plant, next in 
 those on the oat plant ; while those on the corn plant produced young of 
 normal weight and vigor. At the time no definite solution of the observa- 
 tions made was given ; but the possibility of toxic bodies being carried 
 in the rations or produced in the intestinal tract was thought to deserve 
 consideration, as was also the poor mineral content of the diet. 
 
 In 1917 these authors (36) published further work along the same line. 
 In the interim between the two publications, knowledge about the essen- 
 tials of a ration had been expanded by their own work and that of others 
 to include the fat-soluble A and water-soluble B vitamins. 
 
 In their effort to improve the unsatisfactory results with the wheat 
 plant, milk fat was added at the rate of 2 pounds per 100 of the grain 
 mixture. The ration, composed of 6.7 pounds of wheat grain, 0.3 pounds 
 of wheat gluten, plus the milk fat, and 7 pounds of corn stover was given 
 to each of two cows. One of these produced a 46-day premature weak 
 calf that lived 10 hours ; the other produced from the first gestation, a 
 healthy calf 13 days premature ; from the second gestation a weak calf 
 at term which grew strong. These findings were somewhat confusing to 
 interpret at the time and it was concluded that in the previous experi- 
 ments the absence of vitamin A had not been the causal factor in repro- 
 ductive failure. It was considered that the ration was so much improved 
 by the mineral addition through the better roughage used (corn stover 
 in place of wheat straw) and by a more abundant supply of fat-soluble 
 A that successful resistance to the real factor, the toxicity of the wheat 
 kernel, was maintained by the second cow during the first gestation. The 
 toxicity was considered to be accumulative and to show its effect some- 
 what on the offspring from the second gestation. 
 
 This work also showed that rations restricted to the wheat plant did 
 not sustain the growth of Holstein heifers. They failed to come into es- 
 trum and could not be bred. They showed pathological conditions such 
 as blindness, feebleness, and emaciation, and abnormal excitability fol- 
 lowed by collapse. 
 
 In 1920 Hart, McCollum, Steenbock, and Humphrey (37) published 
 further work on the influence of rations restricted to the oat plant on re- 
 production in cattle. At this time they showed that breeding cows were 
 inadequately nourished on this ration and that their offspring were born 
 prematurely, very weak, or dead. In the earlier experiments the oat 
 
10 University of California — Experiment Station 
 
 straw had been grown on an alkaline soil and contained 0.84 per cent 
 calcium oxide, and with this roughage better offspring were produced. 
 In the later work the oat straw contained but 0.47 per cent calcium oxide, 
 and the calves were far inferior. 
 
 Addition of vitamin A to this diet in the form of 2 pounds of milk fat 
 per 100 pounds of grain mixture, or of improved protein by the addi- 
 tion of casein separately or combined did not help reproduction. When 
 calcium salts were added in the form of carbonate, phosphate, or ace- 
 tate, offspring of fair vigor were produced, even without the addition 
 of a better protein or more vitamin A. 
 
 Although this much was accomplished with the experimental ration, 
 the offspring were inferior to those produced from cows fed natural 
 roughage such as corn stover, clover, alfalfa, or even marsh hay grown 
 on an alkaline marsh. It was concluded from these data than an other- 
 wise complete ration for a dry breeding cow should contain at least 0.45 
 per cent of calcium oxide, but that this may not apply when the ration 
 contains some fresh green materials. 
 
 Little was mentioned in this publication regarding toxicity of the 
 plant although just why low calcium intake should be the determining 
 factor in normal or abnormal reproduction was not clear to the workers. 
 They suggested that "on low-calcium rations there can be especially 
 favorable conditions for continual absorption of products of intestinal 
 origin, among which may be bacterial toxins or amines." 
 
 In 1924 Hart, Steenbock, Humphrey, and Hulce (38) reported new 
 observations with reinterpretations of their previous experiments on 
 the nutritive value of the wheat plant. They showed that when yellow 
 corn grain was substituted for the wheat grain on the wheat-plant ra- 
 tion, reproductive failure continued. When the ration of yellow corn, 
 wheat gluten, and wheat straw was supplemented by a salt mixture 
 containing calcium, reproduction was normal. If, however, the original 
 wheat plant ration (wheat grain, wheat gluten, wheat straw) was sup- 
 plemented by the salt mixture containing calcium, nutrition was inter- 
 fered with and there was a disturbance of the reproductive cycle. At the 
 time of these earlier experiments the investigators had not known that 
 yellow corn is liberally supplied with vitamin A, whereas wheat grain 
 contains a very small quantity. They state : "In the light of our modern 
 views of nutrition it is perfectly clear that the wheat ration was deficient 
 in vitamin A and in calcium." They conclude that the addition of these 
 substances makes the wheat-plant ration complete for growth and repro- 
 duction and that therefore the presence of an inherent toxic factor need 
 not be assumed. 
 
Bul. 560] Vitamin- A Deficiency in Range Cattle 11 
 
 While the above classical experiments were being carried out on nu- 
 trition and reproduction, a very large amount of bacteriological investi- 
 gation was conducted on bovine infectious abortion. This latter condi- 
 tion was conclusively demonstrated to be an infectious disease. Natur- 
 ally, however, some conflict of opinion arose between the workers in 
 nutrition and bacteriology on the interrelation of nutritive regimens 
 and specific bacterial infection in reproduction in cattle. 
 
 The discovery by Evans and Bishop (n9) of vitamin E, a hitherto 
 unrecognized dietary factor essential for reproduction, the first note on 
 which appeared in 1922, increased interest in the importance of nutri- 
 tion in reproduction. 
 
 Working on this new information the United States Bureau of Dairy 
 Industry conducted a limited series of experiments in which slight evi- 
 dence was secured that sprouted oats overcame sterility in cows. These 
 animals had been examined previously and showed no evidence of path- 
 ological or diseased condition of the genital tract. The work was pub- 
 lished by Winters (40) in 1926 and by Graves and Miller (41) in 1927. The 
 latter attributed success to the presence of vitamin E in the sprouted 
 oats. Further data on the subject were included by Miller and Graves (42) 
 in a history of reproduction of the Beltsville Herd of the Bureau of 
 Dairy Industry. These meager experiments did not justify the assump- 
 tion made regarding vitamin E and have not been confirmed. 
 
 Infectious abortion in cattle had been given some attention by the 
 National Research Council beginning with 1920. In 1925 the Council 
 appointed a joint committee of the Divisions of Biology and Agriculture 
 and Medical Sciences under the chairmanship of Theobald Smith. The 
 personnel consisted of workers in both the nutritional and bacteriolog- 
 ical fields, with one geneticist. The committee held its first meeting in 
 Chicago June 11 and 12, 1926, and outlined procedures for the further 
 study of nutrition and Brucella abortus separately or in combination as 
 related to reproduction in cattle. 
 
 At this time (1927), from South Africa, Theiler, Green, and Du 
 Toit (43) reported a significant investigation on minimum mineral re- 
 quirements in cattle. These authors state : 
 
 The paper presents a summary of a preliminary series of orientation experiments 
 concerning minimum requirements of growing cattle for calcium, phosphorus, so- 
 dium, potassium, and chlorine. Main attention was concentrated upon phosphorus 
 deficiency in the hope of producing clear clinical cases of "aphosphorosis" for sub- 
 sequent pathological study and comparison with the naturally occurring disease 
 Styfsiekte; but since the question of ratio, of minerals in dietaries has attracted so 
 much attention of recent years, this factor is also considered. The vitamin factors 
 come in for incidental review. 
 
]2 University of California — Experiment Station 
 
 The vitamin factors caused us to review these studies comprising 
 eight duplicate experiments on sixteen heifers. The basal ration con- 
 sisted of 3% pounds of hay, poor in minerals but otherwise of fair qual- 
 ity ; 2 ounces of blood meal ; and 5 pounds or more of Fanko. This last, 
 locally produced, consisted of the rolled endosperm of maize, high in 
 calorific value, fair in protein content, but very low in mineral constit- 
 uents. All the animals used, except those in experiment 8, were on diets 
 low in vitamin A, which was limited to the amount in the 3% pounds of 
 hay. Fanko, it was stated, contained even smaller quantities of vitamins 
 than polished rice, and blood meal also very little. In experiment 8, a 
 check on the vitamin factor, part of the hay was replaced by 4 pounds 
 daily of young, fresh, green forage. 
 
 The experiment, therefore, from the standpoint of this vitamin ar- 
 ranged itself into 2 control animals (experiment 8) and 14 animals (ex- 
 periments 1 to 7) on diets low in vitamin A. The 2 control animals in 
 experiment 8 gave birth to normal calves. Of the 14 heifers low in vita- 
 min A along with other deficiencies, 8 produced weak or dead calves, 2 
 died without having calves, and 4 produced normal calves in June or 
 July, 1926, after having been on the diets from September, 1924. 
 
 The summary leaves open the explanation of the observed abnormali- 
 ties in calving except in the case of phosphorus deficiency, in which defi- 
 nitely abnormal calves may be born. According to present knowledge 
 vitamin A was probably a factor in the failure of reproduction, the 3% 
 pounds of poor-quality ripe hay not furnishing enough for normal re- 
 production in all cases. Similar reproductive failure in sheep fed this 
 ration was reported in two subsequent experiments by Du Toit, Malan, 
 andGroenwald. (44 > 45) 
 
 In 1929 Hadley and Hawn (46) and in 1932 Hart, Hadley, and Hum- 
 phrey^ 7 ' reported experiments of the Wisconsin group following pro- 
 cedures outlined by the committee on abortion of the National Research 
 Council to answer the question : "Is it possible to lower the resistance of 
 cattle by feeding a ration low in both lime and protein so that they be- 
 come more susceptible to contagious-abortion infection?" 
 
 Forty-four Holstein heifers were divided into two lots. Lot 1 received 
 the good ration consisting of alfalfa hay, corn silage, and a grain mix- 
 ture consisting of corn, oats, oil meal, wheat bran, bone meal, and cod- 
 liver oil. Iodized salt was used, and in summer the animals had access to 
 both alfalfa and sweet-clover pasture. Lot 2 received a relatively poor 
 ration consisting of corn silage, timothy hay from acid soils, and a grain 
 mixture of corn, oats, and gluten meal. Common salt was used and in the 
 summer they were pastured on timothy and blue-grass. Both groups did 
 well, and eventually all were bred and calved at term. 
 
Bul. 560] Vitamin- A Deficiency in Range Cattle 13 
 
 They were milked for 3 months, bred again for second calves, and 
 when proved by rectal examination to be pregnant were divided into five 
 groups, each containing approximately the same number of animals 
 from lots 1 and 2. Four of these groups were exposed to infection with 
 the abortion organism while the fifth was left as controls. 
 
 This experiment showed that the good ration did not increase resist- 
 ance to infectious abortion, nor did the poor one increase susceptibility. 
 There were eleven abortions on the good ration; eight on the poor. The 
 experiment also showed that cows could adjust themselves to a low lime 
 intake and that the efficiency of lime utilization increased as the amount 
 in the ration decreased. There was also evidence that cod-liver oil in the 
 good ration depressed milk-fat production. E. B. Hart, who had charge 
 of the nutrition of the animals in this experiment, commented in the dis- 
 cussion of the 1929 report as follows : 
 
 We chose this poor ration because the roughage used was grown on acid soil, which 
 is a condition tliat is common in the northern part of the United States. For milking 
 cows it is no doubt too low in its lime content. The phosphorus content, as Dr. Hadley 
 said, is at a rather low level. If you analyze that and compare it with the alfalfa ra- 
 tion, you are sure to have one distinct deficiency with two others on the border line. 
 The one distinct deficiency in dairy-cow nutrition in the northern states is the use of 
 these poor roughages with their Ioav lime content. You may not know, but it is a fact 
 that with rations still poorer than this poor one that we have, those made from straws 
 and grains, you can disturb reproduction. The deficiency may be lime or it may be one 
 of the vitamins, particularly vitamin A. It was that set-up that led us to this experi- 
 ment. But we put the poor ration as one that is not uncommon on farms. You may 
 say that is not a very poor ration. Indications are that these animals have gotten 
 along very well on it. There has not been much difference in the results from the two 
 rations up to date, it is true. Maybe we will have to revise our idea that the ration 
 we call the poor ration was not so poor after all.( 46 > 
 
 These experiments demonstrated clearly that the specific disease, bo- 
 vine infectious abortion, was not materially influenced by nutrition. 
 Because of the controversial nature of the subject, the results left in the 
 minds of many the idea that restricted nutritional regimens could not 
 interfere with normal reproduction in the bovine. The cited experiments 
 of Hart and his associates, however, had already proved that restricted 
 nutritional regimens involving low lime and vitamin A could prevent 
 normal reproduction. In the experiments on the effect of nutrition on 
 susceptibility to infectious abortion, an abundance of vitamin A was 
 supplied in the poor ration for lot II, while attention was being focused 
 on low calcium. The result showed increased efficiency of calcium utiliza- 
 tion at low levels and indicated that vitamin-A deficiency was the more 
 important cause of the failure of reproduction in the Wisconsin nutri- 
 tion experiments between 1907 and 1924. 
 
14 University of California — Experiment Station 
 
 In 1926 Jones, Eckles, and Palmer (so) showed vitamin A to be an in- 
 dispensable factor in the diet of calves. They concluded that wheat 
 straw is a good source of this vitamin for ruminants. In a calf from a 
 stall-fed cow consuming less than 40 per cent wheat straw in its ration, 
 however, symptoms of vitamin- A deficiency developed. The vitamin-A 
 storage in the liver of calves at birth, which depends on the diet of their 
 dams, was considered to be an important factor. We believe that the 
 amount of the vitamin in the milk, the time of cutting the grain after it 
 has ripened, and the method of handling the straw are also important 
 factors. 
 
 In 1928 Bechdel, Honeywell, and Dutcher (48) studied the effect of 
 feeding five heifers, one to two and a half years of age, a ration deficient 
 in vitamin A. They produced edema in the front legs with declining ap- 
 petite and increased respiratory rate. One animal went completely 
 blind. In two cases of pregnancy the calves were born dead, one being 
 more than two months premature. No statement is made regarding the 
 possibility of abortion infection in these cases. 
 
 Halverson and Sherwood (49) in 1930 reported investigations on the 
 feeding of cottonseed meal to cattle. They showed very conclusively that 
 the so-called poisoning of livestock from cottonseed feeding was due to 
 deficiencies rather than to the gossypol content, which had generally 
 been held to be the cause since Withers and Carruth (50) had isolated this 
 phenol-like substance from the seed and showed it to be toxic for ani- 
 mals. The deficiency primarily responsible for the abnormalities was 
 vitamin A, although vitamin B and calcium were low in the diets and in 
 some cases when yeast and calcium were supplied some improvement 
 was observed. Though these experimental animals showed reproductive 
 difficulties similar to those in the Wisconsin experiments, the data are 
 confusing because infectious abortion existed in some of the cows. Never- 
 theless, the lowered body weight at birth, the weakness and high mor- 
 tality in the offspring, and the presence of eye lesions in the calves from 
 cows on the rations low in vitamin A as compared with the controls are 
 significant. 
 
 In 1932 Meigs and Converse (51) reported the physiological effects of 
 rations containing low-grade roughage on cows free from infectious 
 abortion. After fairly long periods on grain and U. S. No. 3 timothy hay 
 their animals uniformly gave birth to premature dead or weak and blind 
 calves. The hay was graded No. 3 because it had been cut in the seed 
 stage without other damage. When a better grade of timothy was fed to 
 other cows in the same herd the proportion of normal calves was larger ; 
 and when the roughage consisted of U. S. No. 1 alfalfa hay, reproduc- 
 tion was quite satisfactory. 
 
BUL. 560] VlTAMIN-A DEFICIENCY IN RANGE 'CATTLE 15 
 
 Deficiency of vitamin A in the No. 3 hay was considered an important 
 factor in producing the results obtained. Experiments on rats showed 
 that U. S. No. 1 alfalfa contains thirty times as much vitamin A as U. S." 
 No. 3 timothy hay. Even the best timothy hay was found to contain only 
 about one-tenth as much vitamin A as the best alfalfa. 
 
 All critical students of the subject, therefore, must realize that even 
 in cattle vitamin A is a necessary food essential for normal physiolog- 
 ical processes. Reproduction is affected when vitamin A is too low in the 
 diet, and the result is the birth of premature, dead, or weak calves with 
 or without eye lesions, accompanied by retention of the placental mem- 
 branes. These may be the only symptoms noticed. They are so similar to 
 those of infectious abortion of cattle, a widespread infectious disease 
 which is far more likely to be the cause, that possibility of the latter must 
 always be eliminated by bacteriological and serological procedure be- 
 fore the existence of vitamin-A deficiency can be established. All pre- 
 vious evidence of the existence of this condition has been based on the 
 work with experimental groups of cattle on more or less highly restricted 
 nutritional regimens. 
 
 We now desire to present evidence that conditions to which range 
 cattle are more or less regularly subjected may be such that they will 
 not be getting sufficient of this vitamin in their forage and supplements 
 and that severe losses may result. 
 
 EVIDENCE OF VITAMIN-A DEFICIENCY" UNDE& RANGE CONDITIONS 
 
 According to the evidence previously cited, range feed loses its vita- 
 min-A content upon drying and bleaching. Tn the bovine, as in other 
 species, the concentration of vitamin A in the liver varies with feed con- 
 ditions. The reserve supply that has accumulated in the body during 
 the green-feed season, together with the limited additional supply ob- 
 tained in different areas from browse, moss, grass from moist places 
 around streams and waterholes, etc., is sufficient to carry animals 
 through the dry season under average conditions. 
 
 The amount of this substance required by an animal varies with age, 
 rapidity of growth, pregnancy, and lactation. With unusual prolonga- 
 tion of the dry-feed period, therefore, manifestations of the existence 
 of deficiency may occur. During the five-year period in which our range- 
 feed investigations have been carried on, more or less severe losses of 
 calves were reported over wide areas on the range in California. In the 
 fall and winter of 1929-30 and again in 1932-33 these losses were par- 
 ticularly heavy. In these years the feed dried early in the spring, and the 
 
16 University • of California — Experiment Station 
 
 first rains came in late autumn followed by cold weather, so that green 
 feed was not available until January or February of the following year. 
 Under such conditions, the calves are born dead or are weak and die soon 
 after birth. Retention of the placenta by the cows and diarrhea in the 
 weak calves are also common. In general the calves born dead are late in 
 gestation ; but evidence exists that they may be expelled at any time dur- 
 ing the second half of pregnancy. Reports have not been infrequent that 
 as many as one hundred head of pregnant animals have aborted in this 
 way. At the same time stockmen have generally agreed that the losses 
 cease and the remaining pregnant cows calve normally after green for- 
 age has been available for a short time. Blood samples, collected by us 
 and by official veterinarians from a sufficient number of cases in widely 
 scattered herds, have been negative to the abortion agglutination test, so 
 that bovine infectious abortion can be eliminated as the cause of the 
 trouble. Even though some of these herds contained an occasional re- 
 actor to the blood test for abortion, the number of abortions, weak calves, 
 and retained placentae occurring in the nonreactors shows that some 
 other cause besides infectious abortion is operating. 
 
 These manifestations are identical with those described in the con- 
 trolled experiments cited. Further to substantiate the lack of vitamin A 
 in the diet as the cause of this condition, livers of calves were studied 
 chemically by the colorimetric method of Carr and Price (21) during the 
 winter of 1932-33. 
 
 The livers of new-born animals tend to be comparatively low in vita- 
 min A irrespective of the diet of the dam during gestation. Under favor- 
 able conditions of intake, storage accumulates with age and reaches a 
 maximum in mature animals. We have demonstrated that in normal 
 new-born calves solutions representing from 25 to 100 milligrams of their 
 liver tissue are necessary to produce a color reaction with antimony tri- 
 chloride, while as little as 1 milligram of adult cow liver produces the 
 same reaction. Pieces of liver tissue were examined in four aborted calves 
 whose dams were under dry-range conditions in widely separated herds, 
 where heavy losses had been reported. Solutions representing as much as 
 10 grams of liver tissue from these fetuses failed to give any color reac- 
 tion, and this quantity justifies the assumption that no vitamin A was 
 present. In a band of ewes on dry range feed and bean straw about 40 
 head of lambs were born dead. The liver of one brought to" the labora- 
 tory was negative for vitamin A. When the ewes were changed to alfalfa 
 hay, the loss of lambs ceased. Negative results were also obtained with 
 liver samples from swine and poultry on vitamin- A-deficient rations. In 
 chickens and turkeys definite vitamin-A deficiency and high mortality 
 have existed while the color reaction could still be demonstrated in the 
 
Bul. 560] Vitamin- A Deficiency in Range Cattle 17 
 
 livers of the dead fowls. We recognize the nonspecific nature of the color 
 reaction with antimony trichloride. Negative reactions are valuable be- 
 cause they demonstrate that no vitamin A, carotenoids, or any other sub- 
 stances that may produce the reaction are present. From the quantita- 
 tive standpoint positive reactions must be interpreted with caution. In 
 our experience this test has checked sufficiently well with clinical obser- 
 vations to be of value quantitatively even in the absence of spectro- 
 graphic confirmation. 
 
 Despite the apparently large accumulation possible in the livers of 
 adult cattle, if they subsequently go on rations deficient in vitamin A 
 there will be a utilization of reserves and a reduction in the quantity 
 secreted with the milk that can be definitely demonstrated after a period 
 of a few weeks on the deficient ration. Also Halverson and Sherwood (49) 
 showed that cows on sufficiently low intake of the vitamin may manifest 
 symptoms and be at the point of death in about 200 days. Steers fed 
 heavily in dry lot on cottonseed cake and hulls will break with vitamin-A 
 deficiency as soon as 100 to 125 days. Evidently, then, under range condi- 
 tions, reserves could be depleted to the point where reproductive failure 
 would occur without other definite clinical symptoms in the dam. This 
 condition probably results because the rapidly growing fetus in utero 
 is utilizing relatively large amounts of the vitamin. Its greater sus- 
 ceptibility to eye lesions as compared with adult animals is further 
 substantiation. 
 
 NATURALLY OCCURRING VITAMIN-A DEFICIENCY IN EXTREME FORM 
 
 During the past winter we have been able definitely to diagnose vita- 
 min-A deficiency in cattle under natural conditions and to study its va- 
 rious manifestations in animals of different ages. This was possible be- 
 cause weather conditions and management on one ranch in southern 
 Tulare County caused its development in such a severe form that 100 
 animals died and many more showed symptoms in a herd of about 250 
 head. The ranch comprises 3,000 acres, practically all farmed to wheat 
 and barley. The owner maintains 75 head of dairy cows. Calves from 
 these cows, together with others purchased when a few days old from 
 dairy farmers, nurse these animals. The calves are finally sold for veal. 
 
 In the spring of 1932 rains ceased early, so that the feed was dry and 
 the grain fields were yellow by May 15. In the fall the first rains came 
 about the middle of December, followed by such cold weather that green 
 feed did not become available until the middle of February, 1933. 
 
 During this unusually long dry-feed period, covering nine months, 
 part of the cattle grazed the unharvested grain fields through the sum- 
 
IS University of California — Experiment Station 
 
 mer. Thirty-five of the cows were left on a leased adjoining natural 
 range until August 10, when they were placed on stubble from which 
 wheat and barley had been harvested. Early in the fall and through the 
 winter they were fed roughage from a stack adjoining the corral. This 
 consisted of straw mixed with wheat and barley hay showing some traces 
 of green color. The hay came from the borders of the fields that were 
 harvested for grain. Barley was the main concentrate feed. This was 
 later supplemented with whole cottonseed. A group of about fifty steers 
 purchased in October was brought in from native grass range and fed in 
 dry lot. These steers, in addition to the straw, grain hay, and barley, 
 received 2 pounds per head daily of a proprietary concentrate feed con- 
 taining 12 per cent protein. 
 
 Symptoms. — The adult cows manifested less marked deficiency than 
 the young stock. Their general condition was poor, although 6 to 7 
 pounds of barley was fed per head daily in addition to the straw and 
 grain hay. When two parts of cottonseed and one part of barley were 
 substituted for the barley alone there was evidence of improvement, but 
 deficiency symptoms were not relieved. The most interesting symptom 
 was night blindness noticed in the twdlight and after darkness when an 
 electric light was turned on in the corral. If attempts were made to han- 
 dle the cattle at this time they ran into each other or into objects, becom- 
 ing nervous and excitable. A few showed ophthalmia with ulceration of 
 the cornea. Their milk and cream entirely lacked color, as did butter 
 made from the latter. 
 
 These cows started to calve the latter part of August. The early calves 
 were normal and although they became unthrifty, showed no definite 
 symptoms until about 12 weeks of age. Birth of normal calves continued 
 till late in the fall. During December and January, however, when 25 to 
 30 cows calved, all the calves were born weak, developed severe diarrhea, 
 and died when one to five days old. At this same time, calves purchased 
 from dairymen on alfalfa ranches remained normal for six to eight 
 weeks after being brought on the ranch and nursing the same cows. This 
 fact indicated the noninfectious nature of the diarrhea. 
 
 One bull was kept on the ranch. He was in poor condition during the 
 winter, but no data regarding his fertility are available. Ability to serve 
 cows was not completely lost. Many of the cows did not come in estrum 
 for five or six months after calving, when green feed became available. 
 
 The feed-lot steers varied in age from yearlings to two-year-olds. 
 The younger cattle were most affected. Eye lesions were the principal 
 symptom, beginning with profuse lacrymation followed by clouding and 
 ulceration of the cornea. 
 
Bul. 560] 
 
 Vitamin-A Deficiency in Range Cattle 
 
 19 
 
 The young animals, varying in age from a few weeks to one year, 
 showed the most marked and varied symptoms and the mortality was 
 high. The eye lesions varied from profuse lacrymation and slight cloud- 
 ing of the cornea to extensive ulceration, loss of aqueous humor, lense 
 opacity, and shrinking of the eyeball, with complete and permanent 
 blindness. 
 
 The animals exhibited capricious appetites, intermittent diarrhea, 
 general unthrifty appearance and pulmonary complications. With per- 
 iods of inclement weather and exposure, pneumonia was the terminal 
 condition causing death in nearly all cases. 
 
 Fig. 1. — One-year-old steer in advanced stage of vitamin-A defi- 
 ciency. Feeding of cod-liver oil in addition to the feeds it had been re- 
 ceiving resulted in a gradual improvement and a gain in weight of 125 
 pounds in 2 months. 
 
 Liver tissue from four of the dead animals was brought to the lab- 
 oratory and was tested colorimetrically for vitamin A. In all cases 
 solutions representing as much as 10-gram samples were negative. 
 
 On February 28, 1933, three animals were taken to the University 
 Farm. No. 1 was a steer born on the ranch in March, 1932, and weaned 
 September 1. After weaning it was fed straw, grain hay, barley, and 
 salt. It first showed symptoms early in January. On February 28 it was 
 extremely gaunt, totally blind, discharging profusely from the nostrils, 
 very unsteady on its legs, and without appetite (fig. 1) . Drenching with 
 reconstituted dried skim milk was resorted to, and in addition 30 mils of 
 good-quality cod-liver oil was given daily. After four days the appetite 
 began to improve, the discharge from the nostrils ceased, and the muz- 
 zle became moist. The steer was continued on straw, barley, and cotton- 
 
20 University of California — Experiment Station 
 
 seed meal, plus the cod-liver oil, and made a complete recovery except 
 for the loss of sight. 
 
 No. 2, a steer born December 1, 1932, came from an alfalfa ranch and 
 grew normally during the first six weeks. While still nursing it devel- 
 oped severe diarrhea and eye lesions and showed evidence of lung in- 
 volvement on January 19. On February 28, it was extremely emaciated ; 
 the left cornea was badly ulcerated, the right cornea clouded, and res- 
 piration rapid (figs. 2, 3, and 4.) It was selected for pathological study 
 without any thought of its surviving. It died on March 3, and showed 
 extreme lung involvement which had gradually progressed over a 
 period of six or seven weeks to be the final cause of death. 
 
 Fig. 2. — Three-month-old steer in moribund condition from vitamin- A deficiency. 
 It died of pneumonia 2 days after this picture was taken. 
 
 No. 3 was a heifer born on the ranch November 15, 1932, and still 
 nursing its dam. Although it was in nearly normal condition of flesh, 
 the right eye was badly affected with severe ulceration of the cornea 
 (fig. 5). On February 28, its appetite was good, and it was kept on the 
 same diet as at the ranch except that reconstituted dried skim milk was 
 substituted for the milk of its dam. No cod-liver oil was given. It grad- 
 ually became worse, showing emaciation, intermittent diarrhea, irregu- 
 lar appetite, and some evidence of lung involvement by rapid breath- 
 ing under slight exertion. Its regression was very gradual, considering 
 the low vitamin- A intake and the condition on arrival. It was killed 
 March 10 in order to have fresh tissues for pathological study. 
 
Bul. 560] Vitamin- A Deficiency in Range Cattle 
 
 21 
 
 Fig. 3.— i-Lung of steer shown in figure 2. Notice the nodules in the pneumonic 
 area. These contained air and pus and are evidence of the subacute nature of the 
 lung involvement. 
 
 Fig. 4. — Left eye of steer No. 2 removed after death 
 for photographing. It shows the severe ulceration of the 
 cornea. 
 
22 University of California — Experiment Station 
 
 All the animals on the ranch have recovered since the advent of green 
 feed except for blindness in two cases, which will be permanent. This 
 heavy loss could have been obviated entirely had the owner purchased 
 alfalfa hay instead of cottonseed in the fall. 
 
 The feeds on this ranch were inadequate in vitamin A, rather than 
 completely devoid of it. The diet was also relatively low in calcium. The 
 liberal feeding- of grain, coupled with adequate protein when cotton- 
 
 Fig. 5. — Animal No. 3, a heifer, three and one-half months old, showing 
 the typical appearance of the advanced eye lesions in cases of vitamin-A defi- 
 ciency. 
 
 seed was added, stimulated growth and milk production and probably 
 explains the severe incidence of deficiency manifestations. Under nat- 
 ural range conditions with deficient protein and phosphorus and with 
 low energy intake, the manifestations have been limited largely to re- 
 productive difficulties, although the dry range forage is probably 
 equally low in vitamin A. 
 
 It is evident that vitamin-A deficiency in cattle is manifested in a 
 variety of ways, and some of its most outstanding symptoms are such 
 that one must carefully differentiate it from other well-known and 
 widely prevalent diseases. 
 
BUL. 500 J VlTAxMIN-A DEFICIENCY IN RANGE CATTLE 28 
 
 DIFFERENTIAL DIAGNOSIS 
 
 Vitamin-A deficiency in some of its manifestations, may be sympto- 
 matically similar to one of two and possibly three other troubles. Thus 
 the ophthalmia is readily confused with infectious keratitis (pink eye), 
 the abnormalities in reproduction with bovine infectious abortion, and 
 possibly the diarrhea with white scours in calves. Careful observations 
 and, in some cases, laboratory tests, coupled with history and knowledge 
 of conditions under which the animals are being kept will serve to dis- 
 tinguish which trouble is present in most cases. 
 
 Infectious keratitis is a rapidly spreading condition that affects all 
 ages of animals irrespective of condition and feed supply. The infec- 
 tion is commonly transmitted by flies, the conjunctival sac is first in- 
 volved, and clouding of the surface of the cornea quickly ensues. Both 
 eyes are usually affected, and there is general systemic disturbance with 
 fever followed by recovery. Ulcerations of the cornea occur in some cases. 
 
 In ophthalmia from lack of vitamin A, young animals will be af- 
 fected more than mature cows. One eye only is more frequently affected ; 
 the animals are usually emaciated and are on feed lacking the vitamin. 
 Clouding of the cornea may start as small opaque spots in the deeper 
 layers and not cover the entire surface so quickly as in infectious kera- 
 titis. It progresses much more slowly, and ulceration of the cornea is 
 common. There may be complete blindness with no abnormalities of the 
 visible structures of the eye (amarosis) because of involvement of the 
 optic nerve, and night blindness is common. After ulceration has oc- 
 curred it is impossible to differentiate the eye lesion alone from the ul- 
 ceration that sometimes follows infectious keratitis, but the general con- 
 dition of the animal and knowledge of its surroundings will greatly 
 assist in diagnosis. 
 
 The abnormalities of reproduction resemble those of infectious abor- 
 tion, and from field observations alone, when the fetus is aborted, are 
 quite indistinguishable. In range cattle terminating their gestation 
 periods before green feed becomes available, the manifestations are simi- 
 lar to those obtained in the controlled experiments of other investigators 
 as outlined at the beginning of this article. Under these range conditions, 
 however, the cows are subjected to other deficiencies besides vitamin A, 
 of which low phosphorus and low protein are the most outstanding in 
 addition to possible low total energy intake. The actual expulsion of the 
 fetus before it is viable or while it is too weak to live, long attributed to 
 infectious abortion or to some other local cause such as eating acorns or 
 mistletoe, we now believe to result from vitamin-A deficiency. The dif- 
 
24 University of California — Experiment Station 
 
 ferential diagnosis from infectious abortion requires laboratory exami- 
 nation. Blood samples must be taken from the aborting cows and the 
 serum tested in the bacteriological laboratory for the presence of agglu- 
 tinins with Brucella abortus antigen. If any positive reactors are found, 
 infectious abortion is present and must be eliminated before the identity 
 of the vitamin-A deficiency can be established. In testing for vitamin-A 
 deficiency, liver samples from the aborted fetuses or from dead calves 
 are forwarded to the nutrition laboratory to be examined chemically by 
 the colorimetric test. For this purpose samples consisting of about % 
 pound of tissue placed in pint fruit jars must reach the laboratory before 
 decomposition has occurred. Decomposition can best be prevented by 
 chilling or freezing before shipment. A negative chemical test, if the 
 agglutination tests of the cows are negative, indicates that vitamin-A 
 deficiency is causing the trouble ; if the agglutination tests are positive, 
 vitamin-A deficiency may be present along with abortion infection. The 
 diagnosis of vitamin-A deficiency is further strengthened by the finding 
 of ophthalmia in the new-born calves. Though the presence of this condi- 
 tion has been definitely reported on the ranges, we have so far not found 
 it in any of the limited number of cases we have examined. 
 
 Animals in advanced stages of vitamin-A deficiency develop a severe 
 diarrhea. If these are young calves born deficient and getting practic- 
 ally no vitamin A in their mothers' milk, the diarrhea may develop early 
 enough to be confused with white scours. History of the cases and analy- 
 sis of liver tissues may be necessary for positive diagnosis. 
 
 DISCUSSION 
 
 The progress of the authors' studies on range forage as related to nu- 
 trition in animals shows more and more clearly that single deficiencies, 
 uncomplicated by other factors, do not usually occur under natural 
 conditions. We have demonstrated that range forage becomes deficient 
 in phosphorus, protein, and vitamin A. The energy intake may be a lim- 
 iting factor when feed is scarce or when the appetites are diminished by 
 specific deficiencies in the forage. 
 
 Since the rigorous conditions to which animals on unsupplemented 
 range forage may be subjected vary from mild deficiencies to actual 
 starvation, the symptoms vary widely. Some knowledge of these symp- 
 toms, together with information on the feed essentials in supplements, 
 is necessary in order to meet most efficiently and economically the nutri- 
 tive requirements of the animals. 
 
 The ability of cattle to store vitamin A in times of abundance against 
 periods of privation is extremely important under California range con- 
 
Bul. 560 J Vitamin- A Deficiency in Range Cattle 25 
 
 ditions. Thus cottonseed cake, rich in protein and phosphorus, will main- 
 tain cattle on dry forage in thrifty condition as long as sufficient re- 
 serves of vitamin A remain. If, however, the period on forage deficient 
 in vitamin A is extended until the reserves are depleted, this essential 
 then becomes the limiting factor ; and alfalfa hay, green forage, carrots, 
 or other substances containing it must be fed if failure in reproduction 
 or other manifestations of its deficiency are to be avoided. 
 
 According to evidence from varied sources, manifestations of single 
 deficiencies are produced more rapidly by the stimulating effects of an 
 otherwise complete ration. The feeding of cottonseed cake, for example, 
 will result in vitamin-A deficiency after a shorter period than when 
 growth and production are more severely limited by the multiple defi- 
 ciencies of range forage alone. 
 
 Experiments are in progress to determine the period required to de- 
 plete animals of vitamin A after the most ideal conditions for storage 
 and to ascertain the minimum amount of good alfalfa hay necessary to 
 prevent the abnormal termination of gestation. Until more information 
 is available, the range livestock man can do much to prevent losses by 
 observing his animals closely and by so supplementing the range as to 
 keep them in thrifty condition. 
 
 Experiments have clearly shown that when cod-liver oil is incorpo- 
 rated into a finely ground feed, vitamin A is rapidly oxidized. This oc- 
 curs because the thin oil film surrounding the particles presents a rela- 
 tively enormous surface exposed to the air. Obviously, therefore, cod- 
 liver oil or a similar source of vitamin A, mixed in so-called vitamized 
 minerals for animal feeding, would not long retain its potency. Even 
 though it did, the amount ingested with the small amount of mineral 
 consumed would be insignificant in relation to the requirements of the 
 animals. 
 
 When animals are not doing well or when definite losses occur, some 
 reason exists. Bacterial diseases, parasitism, nutrition, and hereditary 
 conditions are all to be considered. Stockmen confronted with these 
 conditions can do their part in adding to the knowledge of the range by 
 reporting their losses to those who are investigating the subject. Fail- 
 ure to do so is often prompted by a feeling that such losses are a personal 
 reflection on the owner, and thus the industry is deprived of the in- 
 creased knowledge that might be obtained through investigation of such 
 reports. 
 
26 University of California — Experiment Station 
 
 CONCLUSIONS 
 
 The existence of vitamin-A deficiency in cattle under natural condi- 
 tions lias been demonstrated. Conditions are present during" the dry- 
 feed season which if unusually prolonged will cause the manifestation of 
 various degrees of this deficiency. The symptoms most commonly seen 
 are as follows : 
 
 1. Birth of dead or weak calves with or without eye lesions and asso- 
 ciated with retention of the placenta. This condition simulates infec- 
 tious abortion. 
 
 2. Severe diarrhea in weak newborn calves which simulates white 
 scours. 
 
 3. Eye lesions, particularly in immature animals, which simulate in- 
 fectious keratitis. 
 
 The manifestation in animals is more severe when the diet is other- 
 wise complete and supplied in amounts above maintenance. Under the 
 multiple deficiencies of the range the manifestations are commonly lim- 
 ited to reproductive failure toward the end of the dry-feed season. 
 
 Data on the nature and occurrence of vitamin A and the experimen- 
 tal evidence on the relation of vitamin A to reproduction have been re- 
 viewed. 
 
 The best practical source of vitamin A now available for livestock 
 feeding is green feed or hay. The vitamin-A potency of hay is associated 
 with the degree of green color. 
 
 ACKNOWLEDGMENT 
 
 The writers wish to express their appreciation to H. C. Jackson, As- 
 sistant Farm Advisor of Tulare County, for help in collecting data on 
 the ranch where the natural outbreak of vitamin-A deficiency occurred. 
 
Bul. 560] Vitamin- A Deficiency in Range Cattle 27 
 
 LITERATURE CITED 
 
 1 Hart, G. H., and H. E. Guilbert. 
 
 1928. Factors influencing percentage calf crop in range herds. California Agr. 
 Exp. Sta. Bul. 458:1-43. 
 
 2 Hart, G. H., H. R. Guilbert, and H. Goss. 
 
 1932. Seasonal changes in the chemical composition of range forage and their 
 relation to nutrition of animals. California Agr. Exp. Sta. Bul. 543:1-62. 
 
 3 McCollum, E. V., and M. Davis. 
 
 1914. Observations on the isolation of the substance in butter which exerts a 
 stimulating influence on growth. Jour. Biol. Chem. 19:245-259. 
 
 * Wachenroder, H. 
 
 1826. Ueber das Oleum radicis Dauci Aetherum, das Carotin, den Carotenzucker 
 und den officinellen suceus Dauci: etc. Diss, de Anthelmintics, Gottingen; 
 Geiger's Magaz. Pharm. 33:144-172. 1831. [Cited by Palmer.] 
 
 s Stokes, G. G. 
 
 1864. On the supposed identity of biliverdin with chlorophyll, with remarks on 
 the constitution of chlorophyll. Roy. Soc. London, Proc. 13:144. [Cited by 
 Palmer.] 
 
 6 Piccalo, G., and Ad. Lieben. 
 
 1866. Giorn. Sci. Nat. Econ. Palermo 2:258-275. [Cited by Palmer.] 
 
 7 Tswett, M. 
 
 1911. Ueber den makro- und mikrochemischen Nachweis des Carotins. Ber. 
 Botan. Ges. 29:630-636. [Cited by Palmer.] 
 
 s Escher, H. H. 
 
 1913. Ueber den Farbstoff des Corpus Luteum. Ztschr. Physiol. Chem. 83:198- 
 211. [Cited by Palmer.] 
 
 9 Palmer, L. S. 
 
 1922. Carotinoids and related pigments. Amer. Chem. Soc. Monograph Series. 
 316 p. The Chem. Cat. Co., New York. 
 
 io Mead, S. W., and W. M. Regan. 
 
 1931. Deficiencies in rations devoid of roughage for calves. Jour. Dairy Sci. 14: 
 283-293. 
 
 ii Steenbock, H., and E. G. Gross. 
 
 1919. Fat-soluble vitamine. II. The fat-soluble vitamine content of roots, to- 
 gether with some observations on their water-soluble vitamine content. 
 Jour. Biol. Chem. 40:501-532. 
 
 i' 2 Steenbock, H., and P. W. Boutwell. 
 
 1920. Fat-soluble vitamine. III. The comparative value of white and yellow 
 maizes. Jour. Biol. Chem. 41:81-96. 
 
 13 Steenbock, H., and E. G. Gross. 
 
 1920. Fat-soluble vitamine. IV. The fat-soluble vitamine content of green plant 
 tissues together with some observations on their water-soluble vitamine 
 content. Jour. Biol. Chem. 41:149-162. 
 
28 University of California — Experiment Station 
 
 j 4 Steenbock, H., M. T. Sell, and P. W. Boutwell. 
 
 1921. Fat-soluble vitamine. V. The vitamine A content of peas in relation to 
 their pigmentation. Jour. Biol. Chem. 47:303-308. 
 is Drummond, J. C. 
 
 1919. Researches on the fat-soluble accessory substance. I. Observations on its 
 nature and properties. Biochem. Jour. 13:81-94. 
 
 i6 Steenbock, H., M. T. Sell, and M. V. Buell. 
 
 1921. Fat-soluble vitamine. VII. The fat-soluble vitamine and yellow pigmenta- 
 tion in animal fats, with some observations on its stability to saponifica- 
 tion. Jour. Biol. Chem. 47:89-109. 
 
 17 Rosenheim, O., and J. C. Drummond. 
 
 1920. On the relation of the lipoehrome pigments to the fat-soluble accessory 
 food factor. Lancet 198:862-864. 
 
 is Drummond, J. C, and A. F. Watson. 
 
 1922. The testing of foodstuffs for vitamins. Analyst 47:341-349. 
 
 19 Drummond, J. C, O. Rosenheim, and K. H. Coward. 
 
 1925. The relation of sterols to vitamin A. Jour. Soc. Chem. Indus. 44:123T- 
 124T. 
 
 20 Rosenheim, O., and J. C. Drummond. 
 
 1925. A delicate colour reaction for the presence of vitamin A. Biochem. Jour. 
 19:753-756. 
 
 2i Carr, F. H., and E. A. Price. 
 
 1926. Colour reactions attributed to vitamin A. Biochem. Jour. 20:497-501. 
 
 22 Euler, B. von, H. von Euler, and H. Hellstrom. 
 
 1928. A- Vitaminwirkung der Lipoehrome. Biochem. Ztschr. 203:370-384. 
 
 23 Moore, T. 
 
 1929. Vitamin A and carotene. Biochem. Jour. 23:1267-1272. 
 
 24 Duliere, W., R. A. Morton, and J. C. Drummond. 
 
 1929. The alleged relation of carotene to vitamin A. Jour. Soc. Chem. Indus. 48: 
 316T-321T. 
 
 25 Moore, T. 
 
 1930. Vitamin A and carotene. V. The absence of liver oil vitamin A from caro- 
 tene. VI. The conversion of carotene to vitamin A in vivo. Biochem. Jour. 
 24:692-702. 
 
 2c Hartman, A. M. 
 
 1931. The vitamin A content of different grades of alfalfa and timothy hays and 
 of hays cured under various conditions. (Abstract of paper read at pro- 
 ceedings.) Jour. Biol. Chem. 92:vii-viii. 
 
 27 Coward, K. H. 
 
 1925. The persistence of vitamin A in plant tissues. Biochem. Jour. 19:500-506. 
 
 28 Drummond, J. C, and K. H. Coward. 
 
 1920. Researches in the fat-soluble accessory factor (vitamin A). VI. Effect of 
 heat and oxygen on the nutritive value of butter. Biochem. Jour. 14:734- 
 739. 
 
 29 Dann, W. J. 
 
 1932. Oxidation of vitamin A in vitro. Influence of the solvent. Biochem. Jour. 
 26:666-678. 
 
Bul. 500] Vitamin- A Deficiency in Range Cattle 29 
 
 so Jones, E. I., C. H. Eckles, and L. S. Palmer. 
 
 1926. The role of vitamin A in the nutrition of calves. Jour. Dairy Sci. 9:119- 
 139. 
 si Moore, T. 
 
 1932. Vitamin A and carotene. Notes on the conversion of carotene to vitamin 
 A in the cow. Biochem. Jour. 26:1-9. 
 s- Bergh, H. H., P. Muller, and J. Broekmeyer. 
 
 1920. Das lipochrome Pigment in Blutserum unci Organen, Zanthosis, Hyper- 
 lipochromamie. Biochem. Ztschr. 108:279-303. 
 as Beach, J. E. 
 
 1924. Studies on a nutritional disease of poultry caused by vitamin A deficiency. 
 California Agr. Exp. Sta. Bul. 378:1-22. 
 34 Hughes, J. S., C. E. Aubel, and H. F. Lienhardt. 
 
 1928. The importance of vitamin A and vitamin C in the ration of SAvine. Kan- 
 sas Agr. Exp. Sta. Tech. Bul. 23:1-47. 
 33 Hart, E. B., E. V. McCollum, H. Steenbock, and G. C. Humphrey. 
 
 1911. Physiological effects on growth and reproduction of rations balanced from 
 restricted sources. Wisconsin Agr. Exp. Sta. Ees. Bul. 17:131-205. 
 
 36 Hart, E. B., E. V. McCollum, H. Steenbock, and G. C. Humphrey. 
 
 1917. Physiological effect on growth and reproduction of rations balanced 
 from restricted sources. Jour. Agr. Ees. 10:175-198. 
 
 37 Hart, E. B., E. V. McCollum, H. Steenbock, and G. C. Humphrey. 
 
 1920. Influence of rations restricted to the oat plant on reproduction in cat- 
 tle. Wisconsin Agr. Exp. Sta. Ees. Bul. 49:1-22. 
 
 38 Hart, E. B., H. Steenbock, G. C. Humphrey, and E. S. Hulce. 
 
 1924. New observations and a reinterpretation of old observations on the nutri- 
 tive value of the wheat plant. Jour. Biol. Chem. 62:317-322. 
 so Evans, H. M., and K. S. Bishop. 
 
 1922. On the existence of a hitherto unrecognized dietary factor essential for 
 reproduction. Science 56:650. 
 40 Winters, S. E. 
 
 1926. Sprouted oats for sterility. Hoard's Dairyman 71:776. 
 4i Graves, E. E., and F. W. Miller. 
 
 1927. Experiments in feeding sprouted oats. Jersey Bul. and Dairy World 46: 
 1251. 
 
 42 Miller, F. W., and E. E. Graves. 
 
 1932. Eeproduction and health records of the Beltsville herd of the Bureau of 
 Dairy Industry. U. S. Dept. Agr. Tech. Bul. 321:1-23. 
 
 43 Theiler, A., H. H. Green, and P. J. du Toit. 
 
 1927. Minimum mineral requirements in cattle. Jour. Agr. Sci. 17:291-314. 
 
 44 du Toit, P. J., A. I. Malan, and S. W. Groenwald. 
 
 1932. Studies in mineral metabolism XVIII. Phosphorus in the nutrition of 
 sheep. 18th Eeport Director of Vet. Services and Animal Industry Union 
 of South Africa, p. 611-630. 
 
 45 du Toit, P. J., A. I. Malan, and S. W. Groenwald. 
 
 1932. Studies in mineral metabolism XX. Iodine in the nutrition of sheep, p. 
 651-676. 
 
30 University of California — Experiment Station 
 
 46 Hadley, F. B., and M. C. Hawn. 
 
 1929. The influence of nutrition on contagious cattle abortion. Proc. 33rd Ann. 
 Meet. U. S. Livestock San. Assoc, p. 308-320. 
 
 *7 Hart, E. B., F. B. Hadley, and G. C. Humphrey. 
 
 1932. The relation of nutrition to contagious cattle abortion. Wisconsin Agr. 
 Exp. Sta. Ees. Bui. 112:1-45. 
 < 8 Bechdel, S. I., H. E. Honeywell, and R, A. Dutches. 
 
 1928. The effect of feeding heifers a ration deficient in vitamin A. Pennsyl- 
 vania Agr. Exp. Sta. Bui. 230:20-21. 
 49 Halverson, J. O., and F. W. Sherwood. 
 
 1930. Investigations in the feeding of cottonseed meal to cattle. North Carolina 
 Agr. Exp. Sta. Tech. Bui. 39:1-158. 
 
 r>o Withers, W. A., and F. E. Carruth. 
 
 1915. Gossypol, the toxic substance in cottonseed meal. Jour. Agr. Research 5: 
 261-288. 
 si Meigs, E. B., and H. T. Converse. 
 
 1932. Physiological effects of rations with low grade roughage. Absts. of papers, 
 27th Ann. Meet, Amer. Dairy Sci. Assoc, p. 55. 
 
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