THE EFFECTS OF VITAMIN DEFICIENCY UPON THE COEFFICIENTS OF DIGESTIBILITY OF PROTEIN,

FAT, AND CARBOHYDRATE

BY R. REDER ST. JULIAN AND VICTOR G. HELLER

(From the Department of Agricultural Chemistry Research, Oklahoma Agri- cultural and Mechanical College, Stillwater)

(Received for publication, July 7, 1930)

The profound metabolic changes which are brought about by a vitamin-deficient diet suggest that the digestive processes may in some way be affected by the lack of these accessory substances. It has been suggested that vitamins may affect the glandular secretory processes connected with digestion or that they may influence the absorption of foods from the alimentary tract and their subsequent oxidation. If the digestive process is affected by a lack of vitamins, we should expect such a disturbance to be evidenced by an inefficient utilization of food.

There has been no work done upon the effect of vitamin A on the utiliza- tion of food except in so far as a vitamin A deficiency is included in condi- tions of polyavitaminosis. Bickel (1) found that although both fat and protein absorption and oxidation are quantitatively elevated during avitaminosis, they are, for the most part, qualitatively normal. Bickel (2) further shows that although during complete avitaminosis the body weight of guinea pigs diminishes and anemia and atrophia changes become promi- nent, the diet is still quantitatively absorbed. He is of the opinion that vitamins do not influence either the digestion or absorption of food ma- terials. He believes that in the absence of vitamins the cells gradually lose their ability to resynthesize and store the food digestion products in the blood and consequently the animal must sacrifice its own tissue.

According to Never (3), the secretory activity and acidity of the gastric juice are not abnormal during avitaminosis; but as the condition pro- gresses, the activity of the pepsin increases and the digestive glands tend to accumulate pepsin and trypsin or their precursors. He further reports that there is a disturbance in absorption and secretion during avitaminosis and that the nitrogen balance is disturbed.

Lavrov and Matzko (4) found that when starvation was prevented in chickens during a B avitaminosis, the utilization of the food nitrogen

99

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100 Vitamin Deficiency Effects

remained satisfactory but that there was a negative nitrogen balance and an increased uric acid excretion which shows that in the initial stages of avitaminosis there is an intense destruction of nitrogenous materials.

Ederer (5) found that vitamin B has a profound influence on protein and carbohydrate assimilation on diets made up of one or of the other of these foodstuffs. Karr (6) found that nitrogen utilization in the digestive tract is unaffected by the lack of the vitamin B complex. In a metabolic study of scurvy, Baumann and Howard (7) report a high negative nitrogen balance during the scorbutic period. Shipp and Zilva (8) explain this as due entirely to an insufficient intake of nitrogen and calories. To over- come this difficulty, Jarussowa (9) forcibly fed animals on a scorbutic diet, so that they maintained their weight for a period of 24 days, during which time they were in positive nitrogen balance. Immediately thereafter the nitrogen balance became negative. She concludes that during the develop- ment of scurvy, the nitrogen balance changes from positive to negative.

Shipp and Zilva, in their study of the effect of vitamin C deficiency upon the metabolism of growing guinea pigs, found no indications of a disturbed absorption or retention of nitrogen during the early stages of the develop- ment of scurvy. The nitrogen balance became negative only when the intake of food was diminished as a consequence of the disease.

There seems to have been no work done upon the effect of vita-

min D upon the digestibility of foods, except as it is included in the investigations by Bickel of metabolism during poly- avitaminosis.

The purpose of the present work was to study the individual effects of vitamins A, B1, Bz, C, and D upon the utilization of foods by determining the coefficients of digestibility of protein, fat, and carbohydrate in animals deprived of the particular vitamin under consideration and in those receiving a complete ration. The term digestibility as used here should be considered as the apparent rather than the real digestibility.

Method

The general plan of the experiment was to keep the animals upon a diet deficient in one of the vitamins until the reserve of that vitamin stored in the animal was entirely depleted. The missing supplement was then added to the diet of part of the animals while their litter mates were continued as controls on the deficient ration. Upon the depletion of the stored vitamin the rats were placed in metabolism cages so that the urine and feces might be collected separately. To avoid a possible misinterpretation of the

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R. R. St. Julian and V. G. Keller 101

results, due to a difference in food consumption in the two groups, the food intake of the animals on the complete diet was in most cases limited to that of the animals on the deficient diet.

The diets were analyzed for protein, fat, moisture, ash, and fiber. The nitrogen of the urine and feces was determined by the Kjeldahl-Gunning method. The fat, crude fiber, moisture, and ash of the feces and diet were determined by the Official Method of Analysis of the Association of Official Agricultural Chemists, The nitrogen balance was determined and the coefficients of digestibility of proteins, fats, and carbohydrates were calculated as follows :

Coefficient of = amount ingested - amount in feces digestibility amount ingested

EXPERIMENTAL

Vitamin A

Twelve rats were evenly distributed in four cages so that the rats in Cages 1 and 3 were litter mates and controls, respectively, of those in Cages 2 and 4. The rats were kept on screens to pre- vent coprophagy. They were fed the following vitamin A-free irradiated ration.

per cent Casein (alcohol-extracted). . . . , . . . . 18 Yeast........................................................ 8 Dextrin...................................................... 56 Saltmixture*................................................ 3 Crisco....................................................... 15

* McCollum’s Salt Mixture 185 (McCollum, E. V., and Simmonds, N., J. Bid. Chem., 33, 63 (1918)).

When losses in weight indicated that the stored vitamin A was exhausted, the rats were transferred to metabolism cages and the collection of urine and feces was begun. Each rat in Cages 2 and 4 was given 0.5 cc. of cod liver oil daily. The animals in Cages 1 and 3 were continued on the deficient ration. The food imake of the rats in Cages 2 and 4 was limited to that of their litter mates in Cages 1 and 3, respectively. However, one of the rats in Cage 3 died, so an allowance was made for this in the food in- take of the rats of Cage 4.

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102 Vitamin Deficiency Effects

The feces and urine were collected in three periods of 3 days each. The feces were dried at 100” immediately upon collection and the urine was preserved with acid.

Table I shows the amount of nitrogen consumed by the animals of each cage during each period of the experiment and the amount of nitrogen occurring in the urine and feces. The nitrogen balance and the coefficients of digestibility of protein, fat, and carbo- hydrate are calculated.

TABLE I

Nitrogen Balance and Coejkients of Digestibility of Protein, Fat, and Carbohydrate in Rats with and without Vitamin A

Experiment I.

Diet

No vitamin A.. I‘ ‘I “

Vitamin A.. . . “ “ . . ‘I “ . . .

No vitamin A.. “ I‘ “ I‘ “ “

Vitamin A.. . . “ “ . . . ‘I “ . . .

c

-

2

1

2

3

4

Nitrogen

III- ‘God take

Output in CO-

P= 2 g 2 Balance effi- pe- ‘G 4 “0 cien

riod > h H ~-

gn. gm. gm. sm. w.

1 1.260.770.180.95+0.31 85 2 1.050.810.180.99 SO.06 83 3 1.260.760.190.95+0.32 83

1 1.260.760.170.93 $0.33 86 2 1.050.70~0.110.81+0.24 I I 3 1.260.70’0.290.99 $0.28

90 77

1 1.080.860.221.08 +O.Ol 82 2 0.860.640.170.81+0.05 80

Fat K&i- ient

zar- ohy- irate oeffi- :ient

97 98 96 99 99 98

98 99 98 98 95 98

96 97 97 98 97 97

96 97 96 98 98 99

It will be observed from Table I that the animals on the vitamin A-deficient diet and those on the complete ration were in positive nitrogen balance except during one period when there was a slight negative balance occurring in one cage of rats on a complete diet, due to the low food intake.

The experiment was repeated with twelve animals as described above, with the exception that collections were made for three periods of 4 days each. A positive nitrogen balance was found in

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R. R. St. Julian and V. G. Heller 103

all the cages for all periods except for one period in one cage of rats which received vitamin A. The rats which received no vita- min A showed the following average coefficients of digestibility: for protein, 82; for fat, 97; and for carbohydrate, 96. For those animals which received vitamin A the average coefficients of digestibility were: 79 for protein, 98 for fat, and 93 for carbo- hydrate.

The average coefficients of digestibility obtained in the two ex- periments are as follows:

No vitamin A Vitamin A

Protein ........................................ 83 83 Fat ............................................ 97 92 Carbohydrate .................................. 97 96

Vitamin A appears to have no effect upon the digestibility of food.

Vitamins Bl and B2

Twelve young rats were evenly distributed in four cages so that each rat had a litter mate in each of the other three cages. They received the following vitamin B-free ration.

pm cent Casein (water-washed). . . . . . . . . . . . . . . . . . . . . . . . . . . 18 Dextrin.................................................... 60 Saltmixture*.............................................. 3.5 Cod liver oil............................................... 3.5 Crisco..................................................... 15

* McCollum’s Salt Mixture 185.

Upon the depletion of the stored vitamin, as evidenced by a loss in body weight, metabolic studies were begun. The rats of Cage 1, serving as controls, were continued upon the deficient ration without the addition of a supplement. Each rat in Cage 2 was given 0.4 cc. of an alcoholic extract of rice polishings daily; in Cage 3 each animal received 0.75 gm. of autoclaved yeast; and in Cage 4, each received 0.75 gm. of whole yeast daily. The rats were allowed to eat the basal diet ad l&turn. Urine and feces were collected for six periods of 4 days each.

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104 Vitamin Deficiency Effects

Table II presents the nitrogen balance and the coefficients of digestibility as determined in this experiment,

TABLE II

Nitrogen Balance and Coeflcients of Digestibility of Protein, Fat, and Carbohydrate in Rats with and without Vitamin B Complex

Experiment I.

Diet

Ko vitamin B . (1 “ “ “ “ “ i‘ “ ‘I . . . . . “ I‘ “ <I “ “

Vitamin B.. . ‘I “ . ‘I “ . “ “ . . . I‘ I‘ . . . . . “ “ . . . . .

Vitamin Bz.. “ “ . . . . . . . . ‘I “ . . . . . . . “ “ . . . . . . “ “ . . . . “ “ . . . . . . .

Vitamin B1 + Bz. “ I‘ “ ‘I . . . ‘I “ ‘I ‘I I‘ “ “ “ . “ ‘I “ “ “ I‘ ‘I ‘I . . . .

: :age x0.

1

2

3

4

_-~---_

gm. gm. gm. gm. !P.

1 0.880.540.150.69 $0.19 83 2 0.740.440.070.51+0.23 91 3 0.670.550.030.58 +0.09 96 4 0.540.540.100.64 -0.10 81 5 0.540.460.050.51+0.04 92 6 0.470.610.040.65 -0.18 91

1 1.120.610.140.75+0.37 87 2 0.800.490.060.55+0.26 93 3 0.780.450.090.54+0.24 89 4 0.890.500.090.59+0.29 87 5 0.940.590.070.66 +0.28 92 6 1.000.600.070.67+0.33 93

1.620.800.421.22+0.40 74 1.380.690.260.95f0.42 81 1.180.630.290.92f0.26 76

1 85 90 2 1.80/1.00~0.261.26+0.53 1.480.730.140.87 +0.61 90 94 3 1.46'0.82~0.150.97 +0.49

1.88'0.86'0.25 1.11 +0.78 90 95

4 87 91 5 1.88~1.04~0.161.20+0.68

1.90/1.10~0.221.32 +0.58 92 94

6 88 95

-

Fat. xfi- ient

hr- ohy- rate oeffi- ient

-

97 91 96 92

94 97 90

94 95 88

98 95 99 98 98 97 98 97 97 97 97 95

94 95 94 94 92 96

91 94 92 94 97 99

90 92 97 90 93 90

It will be observed from Table II that in this experiment during which there was no control of the food intakes, the animals of the control cage showed a slight positive nitrogen balance throughout

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R. R. St. Julian and V. G. Heller 105

the four periods and a negative balance for two periods. The rats receiving either vitamin B1 or B2 showed an increased positive balance over that of the controls, while those animals which were given whole yeast maintained a positive balance noticeably larger than that shown in the other cages.

A similar metabolic study was made on sixteen rats. In this experiment, vitamin B1 was supplied by 40 mg. of activated fullers’ earth per rat per day. The food intake of the animals of all cages was limited to that of the rats which received no vitamin B. Collections of urine and feces were made for four periods of 3 days each. In this experiment the rats of all cages maintained a slight positive nitrogen balance. The average coefficients ob- tained in the two experiments are summarized in Table III.

TABLE III

Average Coeficients of Digestibility

Experiment I

Protein ........................... Fat. .............................. Carbohydrate .....................

Experiment II

Protein ........................... 84 Fat. .............................. Carbohydrate .....................

/

In both experiments the coefficient of digestibility of protein was noticeably lower in those animals which received vitamin Bz alone than in the controls or in those rats which received vitamin B1 or both vitamins B1 and Bz. With this exception, the presence of vitamin B1, Bz, or both B1 and Bz, does not seem to affect the coefficients of digestibility of protein, fat, and carbohydrate.

Vitamin C

In the first experiment, three young guinea pigs were used. They were kept on screens and were given the following vitamin C-free diet.

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106 Vitamin Deficiency Effects

Ground oats................................................. 59 Skimmed milk powder, heated for 2 hrs. at 110”. . . . . . . 30 Butterfat................................................... 10 Sodium chloride.............................................. 1

Within 15 days all of the animals had lost weight and were clearly suffering from scurvy. At this point collection of the urine and feces was begun. The guinea pigs were kept in me-

TABLE IV

Nitrogen Balance and Coeficients of Digestibility of Protein, Fat, and Carbohydrate in Guinea Pigs with and without vitamin C

Experiment I.

Diet

No vitamin C.. . ‘I I‘ “ . . . . . . “ “ “ . . . . . . ‘I “ ‘I . . . . . .

(

--

Vitamin C.. “ “ . . . . . . “ “ . . . . “ I‘ . . . . .

Vitamin C.. . “ “ . . . . “ “ . . “ “ . . . . .

%:

1

2

3

-

-7

I Nitrogen

In- ‘eriod take

Output in

3 1.790.350.83 1.18 +0.61 54 3 1.200.370.69 1.06 +0.14 42 3 0.650.360.520.88 -0.23 21 3 0.020.450.070.52 -0.50

86 63 85 54 73 56

3 1.75.0.34 1.03 1.37 +0.38 41 77 56 3 1.290.350.751.10 +0.18 42 70 54 3 0.670.330.430.76 -0.09 36 83 47 3 1.470.360.721.08 +0.39 51 85 48

3 1.770.421.091.51+0.26 38 3 1.280.370.731.10+0.19 43 3 0.850.390.230.63 +0.22 73 3 1.670.41 1.20 1.60 +0.06 28

81 81 93 76 ,

-

52 57 71 54

Fat meffi :ient

-

t I- <

0

‘

--

:g lrate- oefli- :ient

tabolism cages arranged with a chute into which the animal had to go to reach its food. This arrangement was used to prevent scattering the food over the feces. One of the guinea pigs was continued on the deficient diet in order to serve as a control; each of the remaining animals was given 5 cc. of tomato juice daily. The food consumption of the animals receiving the sup- plement was limited to that of the control for the first two periods

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R. R. St. Julian and 1’. G. Heller 107

of 3 days each. During the remaining two periods they were allowed to eat the diet ad libitum.

Table IV shows the nitrogen balance and the coefficients of digestibility obtained in the experiment.

In the second experiment in which two guinea pigs were used, the urine and feces were collected from the time that the animals were placed on the deficient diet. As soon as scurvy developed each guinea pig was given 10 cc. of tomato juice daily. Collec- tions were made on Guinea Pig 4 for 43 days and on Guinea Pig 5 for 20 days, at the end of which time it died. The average coefficients of digestibility determined for the periods during which the guinea pigs received no vitamin C were as follows: protein, 50; fat, 90; and carbohydrate, 66. For the periods during which the guinea pigs received vitamin C the following coefficients were found: protein, 49; fat, 89; and carbohydrate, 70. Guinea Pig 4 maintained a positive nitrogen balance except during the period immediately following the introduction of vitamin C into the diet. Guinea Pig 5 was in negative balance in all periods except one.

The average coefficients of digestibility observed in the t,wo experiments on the five animals are as follows:

No vitamin C Vitamin C

Protein........................................ 46 46 Fat............................................ 86 85 Carbohydrate.. . . . . 63 62

The low coefficients of digestibility of protein were apparently not due to a lack of vitamin C, for they remained low even after the administration of the tomato juice. The digestibility of foods does not appear to be affected by vitamin C.

Vitamin D

Twelve rats were distributed among four cages so that the rats in Cages 1 and 3 were litter mates respectively, of those in Cages 2 and 4. The rats were kept in the dark and were placed on raised screens to prevent coprophagy. They were fed the following vitamin D-free ration.

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108 Vitamin Deficiency Effects

per cent Yellowcorn.................................................. 76 Wheatgluten................................................ 20 Calciumcarbonate........................................... 3 Sodium chloride............................................. 1

The rats were placed in metabolism cages as soon as x-ray examination of the bones of the right hind leg showed the animals

TABLE V

Nitrogen Balance and Coejkients of Digestibility of Protein, Fat, and Carbohydrate in Rats with and without Vitamin D

Experiment I.

Diet 2%:

No vitamin D. ...... 1 ,‘ “ “ ....... “ ‘I “ ....... “ “ “ .......

Vitamin D .......... 2 “ ‘I ......... “ “ ......... “ “ .........

No vitamin D. ...... 3 i‘ ‘I “ ....... I‘ I‘ “ ....... I< “ “ .......

Vitamin D ........... 4 ‘I ‘I ......... “ “ ......... I( ‘I .........

I Nitrogen

In- eriod take

Output in

P=. pj g pe- .z p z

I I

CO- ‘ij Balance effi-

cient riod 9 R b

----

gm. gm. gm. gm. gm.

1 1.38 1.000.18 1.18 +0.20 89 2 1.561.100.201.30 +0.26 87 3 1.611.010.231.24+0.36 85 4 1.451.030.251.29+0.17 83

1 1.321.080.171.25+0.10 87 2 1.561.100.211.31+0.25 86 3 1.511.100.181.26+0.25 88 4 1.261.040.191.23+0.03 87

1 1.490.970.171.14+0.35 88 2 1.70 1.170.19 1.36 $0.34 89 3 1.711.250.201.45 +0.26 88 4 1.711.020.231.25 $0.46 87

1 1.43 1.020.141.17 $0.26 90 2 1.520.970.161.13+0.39 87 3 1.40 1.030.141.17 +0.23 90 4 1.531.370.151.52+0.01 90

E c

-

Fat :oeffi :ient

84 84 84 84

84 83 85 84

87 89 85 85

88 86 88 89

h- ohy- irate oe5- ient

-

93 93 92 93

92 92 93 92

92 93 92 92

93 92 93 93

to be rachitic. The controls in Cages 1 and 3 were continued on the vitamin D-deficient diet while those in Cages 2 and 4 were given the basal diet which had been irradiated. The food intake of the rats in Cages 2 and 4 was limited to that of their litter mates in Cages 1 and 3, respectively. The feces and urine were

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R. R. St. Julian and V. G. Heller 109

collected in four periods of 3 days each. x-Ray pictures at the end of the experiment indicated that the animals had been cured.

In Table V are found the nitrogen balances and coefficients of digestibility as determined in this experiment.

This experiment was repeated with twelve rats, asdescribed above. The urine and feces were collected in four periods of 3 days each. In this experiment, cod liver oil was the source of vitamin D. Each rat on the complete ration received 0.5 cc. of cod liver oil daily, administered by means of a pipette. The average coefficients of digestibility found in those animals on the vitamin D-deficient diet were as follows: 88 for protein, 83 for fat, and 93 for carbohydrate. For those animals receiving vitamin D the coefficients of digestibility were 89 for protein, 96 for fat, and 93 for carbohydrate.

In Experiment II, the rats which received cod liver oil showed much higher coefficients of digestibility of fat than did the controls. Since no such difference was shown in Experiment I, in which no cod liver oil was given, it is evident that the high coefficient was caused by a loss of oil during its administration.

Average coefficients of digestibility as determined in the two experiments are :

No vitamin D Vitamin D

Protein ........................................ 87 88 Fat ............................................

/ I

84 88 Carbohydrate .................................. 93 92

The coeflicients of digestibility for protein, fat, and carbo- hydrate appear to be the same in those animals which received vitamin D as in those which were on the vitamin D-deficient diet.

SUMMARY

A comparison has been made of the metabolism of animals on a diet deficient in a given vitamin with the metabolism of animals upon the same diet supplemented with the missing vitamin. Such a study has been made with each of the vitamins, vitamins A, B1, Bz, C, and D. In the case of each of these vitamins, the coefficients of digestibility of protein, fat, and carbohydrate were the same in those animals on the deficient ration as in those which

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Vitamin Deficiency Effects

received the complete diet. It would appear unlikely, therefore, that the digestive process is affected by the vitamins studied.

BIBLIOGRAPHY

1. Bickel, A., &o&em. Z., 146, 493 (1924). 2. Bickel, A., Biochem. Z., 166, 251 (1925). 3. Never, H., Arch. ges. Physiol., 219, 554 (1928). 4. Lavrov, B. A., and Matako, S. N., Biochem. Z., 198, 138 (1928). 5. Ederer, S., Biochem. Z., 168, 197 (1925). 6. Karr, W. G., J. Biol. Chem., 44, 277 (1920). 7. Baumann, L., and Howard, C. P., Am. J. Med. SC., 63, 659 (1917). 8. Shipp, H. L., and Zilva, S. S., Biochem. J., 22, 1449 (1928). 9. Jarussowa, N., Biochem. Z., 198. 128 (19281.

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R. Reder St. Julian and Victor G. HellerPROTEIN, FAT, AND CARBOHYDRATECOEFFICIENTS OF DIGESTIBILITY OF

DEFICIENCY UPON THE THE EFFECTS OF VITAMIN

1931, 90:99-110.J. Biol. Chem. 

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