The Relative Vitamin G Content of Alfalfa Hay, Stems, and Leaves from 10 inch and 24 inch Cuttings*

The Relative Vitamin G Content of Alfalfa Hay, Stems, and Leaves from 10 inch and 24 inch Cuttings*

T h e Relative Vitamin G Content of Alfalfa Hay, Stems, and Leaves from 10 inch and 24 inch Cuttings* VICTOR HEIMAN,! J. S. CARVER, AND J. W. COOK Was...

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T h e Relative Vitamin G Content of Alfalfa Hay, Stems, and Leaves from 10 inch and 24 inch Cuttings* VICTOR HEIMAN,! J. S. CARVER, AND J. W. COOK Washington Agricultural Experiment Station, Pullman, Washington (Received for publication January 31, 1938)


ITAMIN G is one of the important nutrients of properly prepared alfalfa leaf meal. The term vitamin G used in this paper pertains to the growth promoting factor and not to the B2 factor as described

paralysis and promoting increased growth in chicks. Norris and co-workers (1934) reported that when alfalfa meal was fed to breeders as a supplement to a diet deficient in vitamin G, hatchability was improved.


8 4 5 6 7 PERCENTAGE OF DRIED SKIM MILK FIG. 1. The relative vitamin-G content of alfalfa products and dried skimmilk.



by Koehn and Elvehjem (1936). Bethke et al (1931) demonstrated the value of alfalfa leaf meal in preventing nutritional •Published as Scientific Paper Number 382, College of Agriculture and Experiment Station, State College of Washington, Pullman, Washington. t N o w Director of Research, Kasco Mills Inc., Waverly, N.Y. [412]

Hunt, Record, and Bethke (1935) showed with rats that alfalfa hay contained much more vitamin G than vitamin B and that the content of these vitamins decreased in amount as the plant matured. In general, the quantity of these vitamins present was correlated with leanness, greenness, and protein content of the plant.


Wilgus, Norris, and Heuser (1936) showed that the average vitamin G content of sun-cured alfalfa meals was found to be 60 percent that of dried skimmilk, whereas the artificially dehydrated meals contained somewhat more than the sun-cured product. These authors also observed that the greatest concentration of vitamin G was found in the more leafy hays. Douglass, Tobiska, and Vail (1933) reported that the vitamin G in the alfalfa studied was comparatively stable and uniform. Later, Vail, Tobiska, and Douglass (1936) showed that there was not a very substantial difference in vitamin G content between varieties of alfalfa and that the use of artificial drying did not materially affect the vitamin G content. It was the purpose of this investigation to determine by the growth response of cockerels the relative vitamin G content of third cutting alfalfa hay, stems, and leaves obtained from plants which were cut when 10 and 24 inches high. PROCEDURE The alfalfa used in this study was grown at the Irrigation Branch Experiment Station, Prosser, Washington. The field from which the alfalfa was cut was typical irrigated farm land of the region, with a heavy stand of alfalfa which had been cut twice before the experimental samples were harvested. Two adjacent one-twentieth acre plots were marked off. One of these was cut when the average height of the alfalfa was 10 inches above the stubble. The alfalfa was mowed and allowed to remain in the swath four hours, after which it was thrown into windrows with a side delivery rake. The windrows were turned at noon the following day and the hay was removed from the field 48 hours from the time of cutting. Part of the hay was passed through a separator which threshed the leaves from the stems. Finally, by passing the threshed hay over a shaker and through


a fanning mill an almost complete separation of stems and leaves was obtained. Samples of hay, stems, and leaves were finely ground and stored in burlap bags for future use. Preparation of the 10-inch alfalfa was started August 5. On August 19 the adjacent plot contained alfalfa 24 inches above a two to three inch stubble. The alfalfa was in the early bud stage. This plot was subjected to the same treatment just described for the 10-inch plants. Table 1 shows the chemical analysis of these alfalfa hay, stem, and leaf samples. TABLE 1.—Chemical analysis on the air-dry basis of alfalfa hay, stems, and leaves from 10 inch and 24 inch cuttings Stage of cutting

Part of Protein the plant

10 inch alfalfa

hay stems leaves

24 inch alfalfa

hay stems leaves



percent percent percent 1.62 0.33 22.5 1.09 0.28 14.9 2.02 0.38 28.9 15.9 10.5 26.1

1.48 0.99 2.58

0.22 0.19 0.29

The relative vitamin G content of the samples of hay, stems, and leaves from 10 and 24 inch alfalfa was determined by using these products as supplements in a chick ration which was deficient in the vitamin G growth factor and comparing the increases in body weight with those obtained by feeding various levels of dried skimmilk as the supplement. In preparation for this experiment 700 day-old White Leghorn cockerels were fed a ration deficient in vitamin G for a preliminary period of two weeks. This ration consisted of the following ingredients: Ingredient Ground yellow flint corn Wheat flour middlings White wheat bran Purified casein Calcium phosphate (tribasic) ppt

Percent 50.0 20.0 5.0 13.4 1.7



Calcium carbonate, ppt Salt Concentrated cod liver oil Granulated sugar Total

0.9 0.5 0.25 8.25 100.00

Crude protein Calcium Phosphorus

20.0 1.0 0.7

The purified casein used in the experimental rations was prepared by the method of Ringrose, Norris, and Heuser (1931). Following this depletion period 360 cockerels, weighing within ± 5 grams of the mean chick weight, were divided into 24 groups of IS chicks each. The experimental rations were then fed to duplicate groups as indicated in Table 2. The alfalfa and milk supplements were included in the depletion ration given above replacing a part of the granulated sugar, and each of these rations was standardized with a protein level of 20.0 percent, calcium 1.0 percent, and phosphorus 0.7 percent. The twoweek depletion period was followed by a three-week experimental period. Details of this technic have been previously described by Heiman and Carver (1937). RESULTS AND DISCUSSION

The results of this investigation are summarized in Table 2. The mean weight of the groups of selected chicks at two weeks was 57 ± 1 gram which indicates the uniformity of the birds at the beginning of the experiment. The mean body weights at the end of the experimental period are given for each group in Table 2 and the mean body weight "by rations" in the adjoining column is determined by combining the results of duplicate groups. The only type of paralysis (Table 2) encountered in this experiment was nutritional paralysis as described by Norris et al (1931) and not the B 4 or encephalomalacia type of paralysis described by Elvehjem, Phillips, and Hart (1937).

Figure 1 shows the line of least squares fitted to the data plotted from the mean body weight of the chicks which were fed 0.0, 2.5, 5.0, and 10.0 percent dried skimmilk as the supplementary source of vitamin G. The increase in the weight of the chicks receiving the alfalfa supplements over the mean weight of the negative control groups was projected on this line of best fit, and by interpolation, the results obtained by feeding the alfalfa products are expressed as equivalents of dried skimmilk. Five percent of the 10 inch cutting of alfalfa hay was equivalent to slightly more than 2.5 percent dried skimmilk in vitamin G. The stems of the 10 inch hay fed at a 5 percent level contained a quantity of vitamin G equivalent to 1.5 percent dried skimmilk, whereas the 5 percent addition of leaves of the 10 inch hay contained a quantity of vitamin G almost equivalent to that of 5.5 percent additions of the dried skimmilk. Likewise, when the leaves were fed at the 2.5 percent level, they contained slightly more vitamin G than 2.5 percent dried skimmilk. In every case, the alfalfa hay from the 24 inch cutting showed an equivalent of 0.5 percent of dried skimmilk less than the alfalfa products from the 10 inch cutting. The above interpretations have been based upon the increase of body weight of the chicks over the control group. An analysis of the growth based upon the feed consumption gives comparable results. Table 2 gives the feed consumed per chick from the second to the fifth week. The amount of supplement that each chick obtained was calculated from the total feed consumed and percentage of supplement in the feed. The last column shows the grams gained in body weight over the control group at five weeks for each gram of supplement consumed. These data show the same relative values for the vitamin G supplements as the above growth data.


VITAMIN G CONTENT OF ALFALFA HAY, STEMS, AND LEAVES TABLE 2.—Summary of data on tody weight, mortality, nutritional {. aralysis, iindfeed consumption

Group No.

Variable in ration

Mean body weight at 5 weeks

Gain in weight over control Morgroup at tality 5 weeks

by by by groups rations rations 1 13

Negative control

grams grams 84 84 84

Feed consumed Nutri- per chick tional from paraly2-5 sis weeks

by by groups groups

Vitamin Grams gained in body G supplement con- weight over sumed per control group at 5 weeks chick per gram of from 2-5 supplement weeks consumed

by groups

by rations

by rations

grams no. cases no. cases grams 0 1 5 143 3 2 143

grams 0.0


2 14

2.5 percent dried skimmilk

108 110



0 0

5 2

205 181



3 15


percent dried skimmilk

135 142



0' 1

7 5

213 253



4 16


percent dried skimmilk

189 195



0 0

0 0

320 317



5 17


percent 10 inch alfalfa hay

111 113



2 0

7 5

205 214



6 18


percent 10 inch alfalfa stems

102 99



1 1

2 2

190 168



7 19


percent 10 inch alfalfa leaves

146 139



0 0

8 10

259 242



8 20

2.5 percent 10 inch alfalfa leaves

112 110



0 0

8 10

218 178



9 21


percent 24 inch alfalfa hay

101 114



1 0

3 3

193 194



10 22


percent 24 inch alfalfa stems

90 97



2 1

1 6

182 202



11 23


percent 24 inch alfalfa leaves

130 144



1 0

5 7

203 241



12 24

2.5 percent 24inch alfalfa leaves

109 106



0 0

3 5

179 181



L.D.' :

10 The least difference in grams between any two means required to give odds of 19:1 (Snedecor, 1934). SUMMARY

1. The relative vitamin G content of alfalfa hay, stems, and leaves from 10 and 24 inch cuttings was compared with dried skimmilk in a growth experiment with White Leghorn cockerels. The 10 inch alfalfa products fed at the same level contained more vitamin G than those prepared from the 24 inch cutting.

2. Giving the vitamin G content of the sample of dried skimmilk used a numerical value of 100, then the vitamin G content of the 10 inch alfalfa hay was 50, the stems 30, and the leaves 110. For the 24 inch hay, the vitamin G content was 40 and for the stems and leaves of this hay values were 20 and 100, respectively. The authors wish to express their thanks



to Mr. H. P. Singleton, Superintendent of the Washington Irrigation Branch Experiment Station, Prosser, Washington, for preparing the alfalfa products used in this work. REFERENCES

Bethke, R. M., P. R. Record, and D. C. Kennard, 1931. A type of nutritional leg paralysis affecting chicks. Poultry Sci. 10:355-368. Douglass, E., J. W. Tobiska, and C. E. Vail, 1933. Studies on changes in vitamin content of alfalfa hay. Colorado Agr. Exp. Sta. Bui. 4. Elvehjem, C. A., P. H. Phillips, and E. B. Hart, 1937. The differentiation between vitamin B4 deficiency and "encephalomalacia" in growing chicks. Proc. Soc. Exp. Biol. Med. 36:129-131. Heiman, V., and J. S. Carver, 1937. The relative flavin (vitamin G) content of dried skimmilk, dried whey, and dried buttermilk. Poultry Sci. 16 :434-437. Hunt, C. H., P. R. Record, and R. M. Bethke, 1935. The effect of stage of maturity and method of curing upon the vitamin B and vitamin G

content of alfalfa, clover, and timothy hays. Jour. Agr. Research. 51:251-2S8. Koehn, C. J., Jr., and C. A. Elvehjem, 1936. Studies on vitamin G (B2) and its relation to canine black tongue. Jour. Nut. 11:67. Norris, L. C , G. F. Heuser, H. S. Wilgus, Jr., and A. T. Ringrose, 1930. The occurrence in chicks of a paralysis of nutritive origin. Poul. Sci. 10:93-97. Norris, L. C , G. F. Heuser, A. T. Ringrose, H. S. Wilgus, Jr., and V. Heiman, 1934. The vitamin G requirement of poultry. 5th Cong. Mondiale di Pollicultura Atti. 2 :S12-S20. Ringrose, A. T., L. C. Norris, and G. F. Heuser, 1931. The occurrence of a pellagra-like syndrome in chicks. Poul. Sci., 10:166-177. Snedecor, G. W., 1934. Calculation and interpretation of analysis of variance and covariance. Collegiate Press, Inc., Ames, Iowa. Vail, C. E., F. W. Tobiska, and E. Douglass, 1936. Further studies on vitamins in alfalfa. Colorado Agr. Exp. Sta. Bui. 18. Wilgus, H. S., Jr., L. C. Norris, and G. F. Heuser, 1936. The relative vitamin G content of alfalfa meal. Poultry Sci. 15:446-453.