- Email: [email protected]
Further Studies on the Riboflavin Requirements of the Chicken*
Further Studies on the Riboflavin Requirements of the Chicken* CHARLES H.
HUNT
Department of Animal Industry, Ohio Agricultural Experiment Station, Wooster A.
R.
WINTER
Department of Poultry Husbandry, Ohio State University, Columbus R.
M.
BETHKE
Department of Animal Industry, Ohio Agricultural Experiment Station, Wooster (Received for publication January 16, 1939)
HE vitamin requirements of chicks and laying hens occupy an important place in poultry nutrition research. Vitamin G has been shown (Norris, Heuser, and Wilgus, 1930; Bethke, Record, and Kennard, 1931 and 1933; Heiman, 193S; Lepkovsky and Jukes, 193S; Hunt, Record, and Bethke, 1936; and others) to be required by the chicken for normal growth and normal reproduction. Bethke, Record, and Wilder (1937) showed that the beneficial results observed when yeast, liver, milk, and so forth, were fed to chicks was in a large measure due to their riboflavin content. Norris and associates (1936); Heuser, Wilgus, and Norris (1938); and Davis, Norris, and Heuser (1938) placed the vitamin G requirements of chicks on a quantitative basis in terms of chick units; while Hunt, Record, and Bethke (19351936) made a similar determination in terms of rat units. More recently, Lepkovsky et al. (1938) presented evidence that riboflavin was of primary importance in sustaining high production of good hatchable eggs. The relation of riboflavin and vitamin G has recently been reviewed by Bethke, Record, and Wilder (1937), so * Published with the permission of the Direc. tor of the Ohio Agricultural Experiment Station, Wooster.
no attempt will be made to repeat it here. Investigations on growth have, for the most part, been carried out with chicks and extended over a period of 6 to 12 weeks. Hatchability studies have in the main been conducted with hens whose histories before laying were unknown or had been reared on a good poultry ration. The present report includes a study of the effect of different levels as well as different natural commonly used sources of ribofiavin-carrying feedstuffs on growth, production, and reproduction and on production and reproduction in range-reared pullets. EXPERIMENTAL PROCEDURE
The experimental studies were divided into two parts. The first part involved the use of range-reared White Leghorn pullets just starting to lay. In the second part, day-old White Leghorn chicks reared on the experimental rations were continued through egg production and hatchability studies. Part One Range-reared pullets from the university farm were placed in individual laying batteries and fed the rations shown in Table 1. Each ration was fed to 12 birds. The dehydrated alfalfa leaf meal and dried
[330]
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T
FURTHER STUDIES ON THE RIBOFLAVIN REQUIREMENTS OF THE CHICKEN
331
TABLE 1.—Laying ration Ration No. 1 2 3 4 5 6
Yellow corn
Ground wheat
60 56.3 52.5 57.1 54.2 53.4
20 20 20 20 20 20
Wheat bran 5 5 5 5 5 5
Argentine casein
Steamed Oyster bone shells meal
8 6.7 5.5 5.9 3.8 4.6
3 3 3 3 3 3
1 1 1 1 1 1
Cod liver oil 1 1 1 1 1 1
Dried skimmilk
Alfalfa leaf meal 5 10
5 10 5
5
The rat growth method was used in assaying the dried eggs and experimental rations. The rats were confined in wire cages provided with screen bottoms when weighing 34-45 grams and fed a vitamin B complex free ration consisting of vitaminfree casein, 20; sucrose, 71; salt mixture (O and M), 4; Crisco, 3; and cod liver oil, 2. The basal ration was supplemented with a daily addenda of 10 micrograms of thiamin (Merck's) and a water soluble fraction of yeast from which the riboflavin had been removed. The amount of yeast extract (ribofiavin-free) fed daily was equivalent to 1.5 gms. dried yeast. This amount of yeast extract when supplemented with adequate riboflavin and thiamin produced excellent growth, 20-25 gms. per week. When the rats had ceased to gain in weight, they were transferred to individual cages and the finely ground feed and dried eggs were fed as the source of riboflavin. The supplemental feeding was continued for six weeks. Pure riboflavin was used as a positive control. A negative control was run with all groups or trials. RESULTS AND DISCUSSION The riboflavin content of the rations and eggs, feed consumption of the birds in terms of hen days, egg production, hatchability of eggs, and livability of chicks are shown in Table 2. The data show that the feed consumed per hen day increased as the total amount of the riboflavin-carrying ingredients
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skimmilk used as supplemental sources of riboflavin were average commercial products. Sufficient quantities of each were obtained to supply the requisite amount for the entire experiment. The rations were all adjusted so that the protein in each was practically the same—15.40 ± 0.11 percent. The feed consumption of each group was kept throughout the length of the experiment of 20 weeks. Egg production records for each bird were recorded for the entire period of the experiment. No eggs were collected for hatching purposes until the pullets had been on the experimental rations for four weeks. The pullets were stud mated to several males which were rotated at frequent intervals. The eggs from each hen were incubated weekly for 16 weeks. Chicks hatched from the different groups of pullets were confined in battery brooders and fed a riboflavin-deficient ration (ration 1, Table 3) to determine whether the amount of riboflavin in the feed of the hen would affect the amount stored in the chick as judged by livability. The experimental rations and the eggs from the different groups of pullets were assayed for riboflavin with rats. The eggs used for assay were collected during the ninth and tenth weeks of the experiment. The whites and yolks were thoroughly mixed and dried in shallow porcelain pans before an electric fan in a dark room. The dried eggs and rations were finely ground and kept at refrigeration temperature.
2 2 2 2 2 2
Salt NaCl*
332
CHARLES H. HUNT, A. R. WINTER AND R. M.
ability, and livability records were those from hens in lots 5 and 6, receiving 10 percent dried skimmilk, and 5 percent each of alfalfa leaf meal and dried skimmilk, respectively. The hatchability results also indicate that the minimum riboflavin requirements for good embryonic development are close to the value found for the rations of lots 5 and 6 (220 to 230 micrograms of riboflavin per 100 grams of feed). The hatchability of the eggs in lot 3 (10 percent alfalfa leaf meal) was below what one might expect from the riboflavin content of the ration. This may be accounted for in part by the fact that there were several hens in this lot that produced eggs of very low hatchability—ranging from 0.0 to 77 percent. The data in Table 2 also show that there is a fair correlation between egg production and the riboflavin intake per hen day or the riboflavin content of the feed. The amount of riboflavin required to produce good hatchable eggs agrees well with the
TABLE 2.—Record of feed consumption and hen performance
Lot no.
Distinguishing feature of rations Basal ration
Feed per hen day gm.
Riboflavin Riboflavin Producper gm. per hen tion ration day
Hatch of fertile eggs
Livahility of chicks from eggs, 4 weeks
pet.
pet.
57
micrograms 0.8
micrograms 45.6
27.8
micrograms 6.4
5 percent dehydrated alfalfa leaf meal
83
1.50
124.5
49.6
9.6
3.8
54.0
3
10 percent dehydrated alfalfa leaf meal
85
1.90
161.5
52.1
14.4
32.0*
78.0
4
5 percent dried skimmilk
81
1.60
129.6
45.5
11.0
15.0
54.0
10 percent dried skimmilk
86
2.30
197.8
60.0
20.0
68.0
77.0
5 percent alfalfa leaf meal plus 5 percent dried skimmilk
98
2.20
215.6
62.8
20.0
73.0
84.0
1
2
5 6
pet.
Riboflavin per gm., eggs air dried
* This lot contained several hens which produced eggs of very low hatchability.
0.0
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(alfalfa leaf meal and dried skimmilk) in the ration were increased. The results of the assay of the eggs show that the micrograms or riboflavin per gram of egg (air dry) varied directly with the riboflavin content of the feed. It was noted that egg production on the riboflavin-deficient ration (lot 1) was normal for several weeks and then declined to zero; whereas the hatchability of the eggs had dropped to zero by the fourth week. This suggests that the riboflavin requirements for hatchability are somewhat greater than for egg production, which is in accord with the observations of Norris and associates (1936); Davis, Norris, and Heuser (1938); and Lepkovsky et al. (1938). The groups of hens which produced eggs of highest riboflavin content also showed the highest hatchability and the chicks from such eggs showed the highest livability. This confirms the work of Heiman (1935) and Lepkovsky and associates (1938). The best egg production, hatch-
BETHKE
FURTHER STUDIES ON THE RIBOFLAVIN REQUIREMENTS OF THE CHICKEN
333
TABLE 3.—Starting ration Ration No.
Yellow corn
Ground wheat
Wheat bran
Argentine casein
Steamed bone meal
1 2 3 4 5 6
58 56.3 50.5 55.1 52.2 51.3
20 20 20 20 20 20
5 5 5 5 5 5
12 10.7 9.5 9.9 7.8 8.7
3 3 3 3 3 3
Part Two Six lots of 40 day-old White Leghorn chicks from the university farm were placed in battery brooders and fed the rations shown in Table 3. The rations were of approximately the same total protein content—18.46 ± 0.11. The chicks were weighed weekly for eight weeks. As soon as the sex of the
Cod liver oil
Dried skimmilk
Alfalfa leaf meal
—
—
•
—
•— 5 10 5
5 10
— — 5
chick could be determined the males were discarded. Feed consumption records were kept of all lots. At the end of the eighth week the pullets were transferred from the brooder to all-purpose batteries for continued growth and egg production. During the period when eggs were being collected for hatching, male birds from the university farm were rotated every few days from battery to battery. Individual egg production and hatchability records were not kept in this trial. RESULTS AND DISCUSSION The data are recorded in Tables 4, 5, and 6. Each table represents a different period in the life cycle of the chicken, namely (1) rapid growth, 1-12 weeks;
TABLE 4.—Record of feed consumption and chick performance—1-12 weeks Lot No.
Distinguishing feature of rations used
Riboflavin Weight Feed per Feed per Riboflavin per gm. per chick of chicks, L i a b ility gm. gain chick day day feed 12 weeks gm.
gm.
micrograms 7.4
gm.
pet.
214±9
86.0
1
Basal ration
5.4
9.3
micrograms 0.8
2
5 percent dehydrated alfalfa leaf meal
4.1
17.8
1.5
26.7
466 + 29
96.0
10 percent dehydrated alfalfa leaf meal
3.6
23.3
1.9
44.3
687 ± 3 6
96.0
3 4
5 percent dried skimmilk
3.3
22.3
1.6
35.7
685 + 23
100.0
5
10 percent dried skimmilk
3.4
23.4
2.3
53.8
700+19
95.0
3.5
22.5
2.2
49.5
618 + 20
99.0
6
5 percent dehydrated alfalfa leaf meal-)5 percent dried skimmilk
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findings of Norris et al. (1936), and Davis, Norris, and Heuser (1938), while the amount of riboflavin necessary for good egg production is higher than the Cornell workers but is in agreement with the findings of Lepovsky et al. (1938) in that a high riboflavin containing feed is necessary for good production.
Salt NaCl
334
CHARLES H. HUNT, A. R. WINTER AND R. M.
(2) slow growth, 13-20 weeks; and (3) egg production, 21-40 weeks. The data presented in Table 4 show that when alfalfa leaf meal was used as the source of supplemental riboflavin the feed requirements per gram of gain decreased and the feed consumed per chick day increased as the amount of leaf meal in the ration increased. The rations containing
BETHKE
feed consumption per chick day increased as the amount of alfalfa leaf meal and therefore the amount of riboflavin in the ration increased. It is again apparent, as in Table 4, that the same was not true in case of the dried skimmilk lots (4 and 5). The reason is not apparent, in this period, as to why the feed consumed per chick day in the groups (Table 5, lot 6) receiv-
TABLE 5.—Record of feed consumption and pullet performance—13-20 weeks Lot no.
Distinguishing feature of rations used
Feed Riboflavin Riboflavin Weight of Feed per chick per gm. per chick per gm. pullets, day gain 20 weeks feed day gm.
10.9
19.9
micrograms 0.8
S percent dehydrated alfalfa leaf meal
8.7
42.4
10 percent dehydrated alfalfa leaf meal
8.6
4
5 percent dried skimmilk
5 6
1 2 3
micrograms 15.9
gm.
pet.
383 ±25
35
1.50
63.6
868 + 46
71
46.1
1.90
87.6
1042 + 45
92
7.0
52.0
1.60
82.2
1203 + 27
76
10 percent dried skimmilk
8.2
50.9
2.30
117.0
1154 + 30
62
5 percent dehydrated alfalfa leaf meal+ 5 percent dried skimmilk
6.8
52.0
2.20
114.4
1089 + 41
77
Basal ration
dried skimmilk as a source of riboflavin did not show a similar decrease in feed requirements, although the requirements in both lots (4 and 5) were low. The chicks receiving alfalfa leaf meal as the source of riboflavin in the ration showed a close correlation between body weight and the riboflavin consumed per gram of feed or per chick day. The livability of the chicks in all groups, except the basal ration, are not significantly different. The data indicate that under the experimental conditions approximately 190 to 200 micrograms of riboflavin per 100 grams of feed were required for maximum growth to 12 weeks of age. The results from the thirteenth to the twentieth week (Table 5) show that the
ing 5 percent each of alfalfa leaf meal and dried skimmilk is the same as the group (Table 5, lot 4) receiving 5 percent of dried skimmilk. The results are not correlated with the riboflavin content of the feed as is the case in lots 2 and 3 (Table S) and is not similar to the results obtained in part one, Table 2. A similar apparent discrepancy appeared in the next 20-week period (Table 6) which will be discussed later. Table S also shows that the riboflavin consumed per chick day is closely correlated with the body weight and livability of the pullet when alfalfa leaf meal is the source of the riboflavin. No such correlation as noted above is shown when dried skimmilk is the supplement (see Table 6 for a possible explanation).
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gm.
Livability
FURTHER STUDIES ON THE RIBOFLAVIN REQUIREMENTS OF THE CHICKEN
Beginning with the twenty-first week, the pullets were given the rations shown in Table 1. The data for the twenty-first to the fortieth week are presented in Table 6. The feed and the riboflavin consumed per hen day in those lots receiving alfalfa leaf meal is closely correlated with production (percentage) and hatchability (percentage) of the fertile eggs set. A similar
335
The fact that the hens receiving 10 percent alfalfa leaf meal (Table 6, lot 3) produced eggs of good (62.8 percent) hatchability, while the hens in part one (Table 2, lot 3) on a similar ration produced eggs of average low hatchability, appear to confirm the statement that this difference may be accounted for on the basis that a number of the hens in the previous ex-
TABLE 6.—Record of feed consumption and hen performance—21-40 weeks Lot No.
Distinguishing feature of rations used
Flavin per Feed Riboflavin Riboflavin Produc- gm. eggs, Hatch ol per hen per gm. per hen fertile tion air dried day day feed eggs micrograms
pet.
1
Basal ration
40
micrograms 0.80
2
5 percent dehydrated alfalfa leaf meal
89
1.50
133.5
34.5
10.0
35.6
10 percent dehydrated alfalfa leaf meal
94
1.90
178.6
51.0
20.0
62.8
3
micrograms 32.0
pet. 0.0
4
5 percent dried skimmilk
84
1.60
134.4
45.0
11.0
9.2
5
10 percent dried skimmilk
85
2.30
195.5
44.7*
17.0
48.1
78
2.20
171.6
48.0*
16.0
72.9
6
5 percent dried skimmilk+ 5 percent dehydrated alfalfa leaf meal
* High range paralysis in these lots.
correlation is not shown in those lots receiving 10 percent dried skimmilk or 5 percent each of alfalfa leaf meal and dried skimmilk in their ration. From previous experiments (part one, table 2) and from the riboflavin content of the ration, the feed per hen day (Table 6, lots 5 and 6) is lower than expected, indicating that an abnormal condition existed in these lots. The fact that range paralysis developed in these lots during this period not only may account for the low feed consumption and low egg production but may also have been an (inherent) inhibiting factor against the predictable effect on growth, based on the riboflavin consumption in the two previous periods as shown in Tables 4 and 5.
periment (Table 2) produced eggs whose embryos were inherently weak. This confirms the findings of Davis, Norris, and Heuser (1938) with reference to the variation in hatchability of eggs from different hens due to difference in inheritance. It is interesting to note that pullets reared on the basal ration did not produce any eggs; whereas pullets reared on a good ration and then transferred to the basal ration (part one) produced normally for a few weeks and then gradually declined. This indicates that the hen stores some riboflavin which is utilized during periods when riboflavin deficient rations are used. It is an interesting fact that the hatchability of eggs from hens receiving
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gm.
336
CHARLES H. HUNT, A. R. WINTER AND R. M. BETHKE
5 percent each of alfalfa leaf meal and dried skimmilk were consistently higher than the eggs from hens receiving 10 percent of dried skimmilk, both rations having about the same riboflavin content. This indicates that a combination of alfalfa leaf meal and dried skimmilk has some factor other than riboflavin which effects embryonic development. Whether these findings can be explained on the basis of the observations of Bauernfeind et al. (1938) that another unknown factor is involved in embryonic development, requires further study.
The amount of feed consumed per hen day increased as the amount of riboflavincarrying ingredients in the ration is increased. The riboflavin content of the egg varied directly with the riboflavin content of the ration of the hen. Eggs of the highest riboflavin content gave the highest hatchability and the chicks from such eggs showed the highest livability when fed a ration deficient in riboflavin. The hatchability results indicate that approximately 220-230 micrograms of riboflavin per 100 grams of feed are required for the production of good hatchable eggs. The data also suggest that the riboflavin requirements for/ good egg production are somewhat less than for maximum hatchability. / The needs Joi riboflavin for maximum growth in chicks to 12 weeks of age are about 190-2(30 micrograms per 100 grams of feed. /
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t
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REFERENCES
Bauernfeind, J. C , A. E. Schumacher, A. Z. Hodson, L. C. Norris, and G. F. Heuser, 1938. A new factor required for growth and reproduction in the domestic fowl. Proc. Soc. Exp. Biol, and Med. 39:108-111. Bethke, R. M., P. R. Record, and D. C. Kennard, 1931. A type of nutritional leg paralysis affecting chicks. Poul. Sci. 10:355-368. , 1933. Relation of the vitamin G complex to hatchability and nutritive value of eggs. Poul. Sci. 12:332-333. , P. R. Record, and O. H. M. Wilder, 1937. Further studies on vitamin G in chick nutrition. Poul. Sci. 16:175-182. Davis, H. J., L. C. Norris, and G. F. Heuser, 1938 The role of vitamin G in reproduction in poultry. Poul. Sci. 17:81-86. , 1938. Further evidence on the amount of vitamin G required for reproduction in poultry. Poul. Sci. 17:87-93. Heiman, V., 1935. Relative vitamin G contents of dried whey and dried skimmilk. Poul. Sci. 14:137-146. Heuser, G. F., H. S. Wilgus, Jr., and L. C. Norris, 1938. The quantitative vitamin G requirements of chicks. Poul. Sci. 17:105-108. Hunt, Chas. H., P. R. Record, and R. M. Bethke, 1935. Effect of the stage of maturity and method of curing upon the vitamin B and vitamin G content of alfalfa, clover, and timothy hays. J. Agr. Res. 5:251-258. , 1936. Observation on the vitamin G requirement of the chicken. Ohio Agr. Exp. Sta. Bimonthly Bulletin 21:127-130. Lepkovsky, S., and T. H. Jukes, 1935. The vitamin G requirement of the chick. J. Biol. Chem. 111:119-131.
Lepkovsky, S., L. W. Taylor, T. H. Jukes, and H. J. Almquist, 1938. The effect of riboflavin and the filtrate factor on egg production and hatchability. Hilgardia 11:559-591. Norris, L. C , G. F. Heuser, and H. S. Wilgus, Jr., 1930. Is the chief value in milk for feeding poultry due to the presence of a new vitamin? Poul. Sci. 9:133. Norris, L. C , H. S. Wilgus, Jr., A. T. Ringrose, V. Heiman, and G. F. Heuser, 1936. The vitamin G requirement of poultry. Cornell Agr. v / E x p . Sta. Bulletin 660.
&
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SUMMARY AND CONCLUSION
/
HUNT
Department of Animal Industry, Ohio Agricultural Experiment Station, Wooster A.
R.
WINTER
Department of Poultry Husbandry, Ohio State University, Columbus R.
M.
BETHKE
Department of Animal Industry, Ohio Agricultural Experiment Station, Wooster (Received for publication January 16, 1939)
HE vitamin requirements of chicks and laying hens occupy an important place in poultry nutrition research. Vitamin G has been shown (Norris, Heuser, and Wilgus, 1930; Bethke, Record, and Kennard, 1931 and 1933; Heiman, 193S; Lepkovsky and Jukes, 193S; Hunt, Record, and Bethke, 1936; and others) to be required by the chicken for normal growth and normal reproduction. Bethke, Record, and Wilder (1937) showed that the beneficial results observed when yeast, liver, milk, and so forth, were fed to chicks was in a large measure due to their riboflavin content. Norris and associates (1936); Heuser, Wilgus, and Norris (1938); and Davis, Norris, and Heuser (1938) placed the vitamin G requirements of chicks on a quantitative basis in terms of chick units; while Hunt, Record, and Bethke (19351936) made a similar determination in terms of rat units. More recently, Lepkovsky et al. (1938) presented evidence that riboflavin was of primary importance in sustaining high production of good hatchable eggs. The relation of riboflavin and vitamin G has recently been reviewed by Bethke, Record, and Wilder (1937), so * Published with the permission of the Direc. tor of the Ohio Agricultural Experiment Station, Wooster.
no attempt will be made to repeat it here. Investigations on growth have, for the most part, been carried out with chicks and extended over a period of 6 to 12 weeks. Hatchability studies have in the main been conducted with hens whose histories before laying were unknown or had been reared on a good poultry ration. The present report includes a study of the effect of different levels as well as different natural commonly used sources of ribofiavin-carrying feedstuffs on growth, production, and reproduction and on production and reproduction in range-reared pullets. EXPERIMENTAL PROCEDURE
The experimental studies were divided into two parts. The first part involved the use of range-reared White Leghorn pullets just starting to lay. In the second part, day-old White Leghorn chicks reared on the experimental rations were continued through egg production and hatchability studies. Part One Range-reared pullets from the university farm were placed in individual laying batteries and fed the rations shown in Table 1. Each ration was fed to 12 birds. The dehydrated alfalfa leaf meal and dried
[330]
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T
FURTHER STUDIES ON THE RIBOFLAVIN REQUIREMENTS OF THE CHICKEN
331
TABLE 1.—Laying ration Ration No. 1 2 3 4 5 6
Yellow corn
Ground wheat
60 56.3 52.5 57.1 54.2 53.4
20 20 20 20 20 20
Wheat bran 5 5 5 5 5 5
Argentine casein
Steamed Oyster bone shells meal
8 6.7 5.5 5.9 3.8 4.6
3 3 3 3 3 3
1 1 1 1 1 1
Cod liver oil 1 1 1 1 1 1
Dried skimmilk
Alfalfa leaf meal 5 10
5 10 5
5
The rat growth method was used in assaying the dried eggs and experimental rations. The rats were confined in wire cages provided with screen bottoms when weighing 34-45 grams and fed a vitamin B complex free ration consisting of vitaminfree casein, 20; sucrose, 71; salt mixture (O and M), 4; Crisco, 3; and cod liver oil, 2. The basal ration was supplemented with a daily addenda of 10 micrograms of thiamin (Merck's) and a water soluble fraction of yeast from which the riboflavin had been removed. The amount of yeast extract (ribofiavin-free) fed daily was equivalent to 1.5 gms. dried yeast. This amount of yeast extract when supplemented with adequate riboflavin and thiamin produced excellent growth, 20-25 gms. per week. When the rats had ceased to gain in weight, they were transferred to individual cages and the finely ground feed and dried eggs were fed as the source of riboflavin. The supplemental feeding was continued for six weeks. Pure riboflavin was used as a positive control. A negative control was run with all groups or trials. RESULTS AND DISCUSSION The riboflavin content of the rations and eggs, feed consumption of the birds in terms of hen days, egg production, hatchability of eggs, and livability of chicks are shown in Table 2. The data show that the feed consumed per hen day increased as the total amount of the riboflavin-carrying ingredients
Downloaded from http://ps.oxfordjournals.org/ at NERL on June 2, 2015
skimmilk used as supplemental sources of riboflavin were average commercial products. Sufficient quantities of each were obtained to supply the requisite amount for the entire experiment. The rations were all adjusted so that the protein in each was practically the same—15.40 ± 0.11 percent. The feed consumption of each group was kept throughout the length of the experiment of 20 weeks. Egg production records for each bird were recorded for the entire period of the experiment. No eggs were collected for hatching purposes until the pullets had been on the experimental rations for four weeks. The pullets were stud mated to several males which were rotated at frequent intervals. The eggs from each hen were incubated weekly for 16 weeks. Chicks hatched from the different groups of pullets were confined in battery brooders and fed a riboflavin-deficient ration (ration 1, Table 3) to determine whether the amount of riboflavin in the feed of the hen would affect the amount stored in the chick as judged by livability. The experimental rations and the eggs from the different groups of pullets were assayed for riboflavin with rats. The eggs used for assay were collected during the ninth and tenth weeks of the experiment. The whites and yolks were thoroughly mixed and dried in shallow porcelain pans before an electric fan in a dark room. The dried eggs and rations were finely ground and kept at refrigeration temperature.
2 2 2 2 2 2
Salt NaCl*
332
CHARLES H. HUNT, A. R. WINTER AND R. M.
ability, and livability records were those from hens in lots 5 and 6, receiving 10 percent dried skimmilk, and 5 percent each of alfalfa leaf meal and dried skimmilk, respectively. The hatchability results also indicate that the minimum riboflavin requirements for good embryonic development are close to the value found for the rations of lots 5 and 6 (220 to 230 micrograms of riboflavin per 100 grams of feed). The hatchability of the eggs in lot 3 (10 percent alfalfa leaf meal) was below what one might expect from the riboflavin content of the ration. This may be accounted for in part by the fact that there were several hens in this lot that produced eggs of very low hatchability—ranging from 0.0 to 77 percent. The data in Table 2 also show that there is a fair correlation between egg production and the riboflavin intake per hen day or the riboflavin content of the feed. The amount of riboflavin required to produce good hatchable eggs agrees well with the
TABLE 2.—Record of feed consumption and hen performance
Lot no.
Distinguishing feature of rations Basal ration
Feed per hen day gm.
Riboflavin Riboflavin Producper gm. per hen tion ration day
Hatch of fertile eggs
Livahility of chicks from eggs, 4 weeks
pet.
pet.
57
micrograms 0.8
micrograms 45.6
27.8
micrograms 6.4
5 percent dehydrated alfalfa leaf meal
83
1.50
124.5
49.6
9.6
3.8
54.0
3
10 percent dehydrated alfalfa leaf meal
85
1.90
161.5
52.1
14.4
32.0*
78.0
4
5 percent dried skimmilk
81
1.60
129.6
45.5
11.0
15.0
54.0
10 percent dried skimmilk
86
2.30
197.8
60.0
20.0
68.0
77.0
5 percent alfalfa leaf meal plus 5 percent dried skimmilk
98
2.20
215.6
62.8
20.0
73.0
84.0
1
2
5 6
pet.
Riboflavin per gm., eggs air dried
* This lot contained several hens which produced eggs of very low hatchability.
0.0
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(alfalfa leaf meal and dried skimmilk) in the ration were increased. The results of the assay of the eggs show that the micrograms or riboflavin per gram of egg (air dry) varied directly with the riboflavin content of the feed. It was noted that egg production on the riboflavin-deficient ration (lot 1) was normal for several weeks and then declined to zero; whereas the hatchability of the eggs had dropped to zero by the fourth week. This suggests that the riboflavin requirements for hatchability are somewhat greater than for egg production, which is in accord with the observations of Norris and associates (1936); Davis, Norris, and Heuser (1938); and Lepkovsky et al. (1938). The groups of hens which produced eggs of highest riboflavin content also showed the highest hatchability and the chicks from such eggs showed the highest livability. This confirms the work of Heiman (1935) and Lepkovsky and associates (1938). The best egg production, hatch-
BETHKE
FURTHER STUDIES ON THE RIBOFLAVIN REQUIREMENTS OF THE CHICKEN
333
TABLE 3.—Starting ration Ration No.
Yellow corn
Ground wheat
Wheat bran
Argentine casein
Steamed bone meal
1 2 3 4 5 6
58 56.3 50.5 55.1 52.2 51.3
20 20 20 20 20 20
5 5 5 5 5 5
12 10.7 9.5 9.9 7.8 8.7
3 3 3 3 3 3
Part Two Six lots of 40 day-old White Leghorn chicks from the university farm were placed in battery brooders and fed the rations shown in Table 3. The rations were of approximately the same total protein content—18.46 ± 0.11. The chicks were weighed weekly for eight weeks. As soon as the sex of the
Cod liver oil
Dried skimmilk
Alfalfa leaf meal
—
—
•
—
•— 5 10 5
5 10
— — 5
chick could be determined the males were discarded. Feed consumption records were kept of all lots. At the end of the eighth week the pullets were transferred from the brooder to all-purpose batteries for continued growth and egg production. During the period when eggs were being collected for hatching, male birds from the university farm were rotated every few days from battery to battery. Individual egg production and hatchability records were not kept in this trial. RESULTS AND DISCUSSION The data are recorded in Tables 4, 5, and 6. Each table represents a different period in the life cycle of the chicken, namely (1) rapid growth, 1-12 weeks;
TABLE 4.—Record of feed consumption and chick performance—1-12 weeks Lot No.
Distinguishing feature of rations used
Riboflavin Weight Feed per Feed per Riboflavin per gm. per chick of chicks, L i a b ility gm. gain chick day day feed 12 weeks gm.
gm.
micrograms 7.4
gm.
pet.
214±9
86.0
1
Basal ration
5.4
9.3
micrograms 0.8
2
5 percent dehydrated alfalfa leaf meal
4.1
17.8
1.5
26.7
466 + 29
96.0
10 percent dehydrated alfalfa leaf meal
3.6
23.3
1.9
44.3
687 ± 3 6
96.0
3 4
5 percent dried skimmilk
3.3
22.3
1.6
35.7
685 + 23
100.0
5
10 percent dried skimmilk
3.4
23.4
2.3
53.8
700+19
95.0
3.5
22.5
2.2
49.5
618 + 20
99.0
6
5 percent dehydrated alfalfa leaf meal-)5 percent dried skimmilk
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findings of Norris et al. (1936), and Davis, Norris, and Heuser (1938), while the amount of riboflavin necessary for good egg production is higher than the Cornell workers but is in agreement with the findings of Lepovsky et al. (1938) in that a high riboflavin containing feed is necessary for good production.
Salt NaCl
334
CHARLES H. HUNT, A. R. WINTER AND R. M.
(2) slow growth, 13-20 weeks; and (3) egg production, 21-40 weeks. The data presented in Table 4 show that when alfalfa leaf meal was used as the source of supplemental riboflavin the feed requirements per gram of gain decreased and the feed consumed per chick day increased as the amount of leaf meal in the ration increased. The rations containing
BETHKE
feed consumption per chick day increased as the amount of alfalfa leaf meal and therefore the amount of riboflavin in the ration increased. It is again apparent, as in Table 4, that the same was not true in case of the dried skimmilk lots (4 and 5). The reason is not apparent, in this period, as to why the feed consumed per chick day in the groups (Table 5, lot 6) receiv-
TABLE 5.—Record of feed consumption and pullet performance—13-20 weeks Lot no.
Distinguishing feature of rations used
Feed Riboflavin Riboflavin Weight of Feed per chick per gm. per chick per gm. pullets, day gain 20 weeks feed day gm.
10.9
19.9
micrograms 0.8
S percent dehydrated alfalfa leaf meal
8.7
42.4
10 percent dehydrated alfalfa leaf meal
8.6
4
5 percent dried skimmilk
5 6
1 2 3
micrograms 15.9
gm.
pet.
383 ±25
35
1.50
63.6
868 + 46
71
46.1
1.90
87.6
1042 + 45
92
7.0
52.0
1.60
82.2
1203 + 27
76
10 percent dried skimmilk
8.2
50.9
2.30
117.0
1154 + 30
62
5 percent dehydrated alfalfa leaf meal+ 5 percent dried skimmilk
6.8
52.0
2.20
114.4
1089 + 41
77
Basal ration
dried skimmilk as a source of riboflavin did not show a similar decrease in feed requirements, although the requirements in both lots (4 and 5) were low. The chicks receiving alfalfa leaf meal as the source of riboflavin in the ration showed a close correlation between body weight and the riboflavin consumed per gram of feed or per chick day. The livability of the chicks in all groups, except the basal ration, are not significantly different. The data indicate that under the experimental conditions approximately 190 to 200 micrograms of riboflavin per 100 grams of feed were required for maximum growth to 12 weeks of age. The results from the thirteenth to the twentieth week (Table 5) show that the
ing 5 percent each of alfalfa leaf meal and dried skimmilk is the same as the group (Table 5, lot 4) receiving 5 percent of dried skimmilk. The results are not correlated with the riboflavin content of the feed as is the case in lots 2 and 3 (Table S) and is not similar to the results obtained in part one, Table 2. A similar apparent discrepancy appeared in the next 20-week period (Table 6) which will be discussed later. Table S also shows that the riboflavin consumed per chick day is closely correlated with the body weight and livability of the pullet when alfalfa leaf meal is the source of the riboflavin. No such correlation as noted above is shown when dried skimmilk is the supplement (see Table 6 for a possible explanation).
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gm.
Livability
FURTHER STUDIES ON THE RIBOFLAVIN REQUIREMENTS OF THE CHICKEN
Beginning with the twenty-first week, the pullets were given the rations shown in Table 1. The data for the twenty-first to the fortieth week are presented in Table 6. The feed and the riboflavin consumed per hen day in those lots receiving alfalfa leaf meal is closely correlated with production (percentage) and hatchability (percentage) of the fertile eggs set. A similar
335
The fact that the hens receiving 10 percent alfalfa leaf meal (Table 6, lot 3) produced eggs of good (62.8 percent) hatchability, while the hens in part one (Table 2, lot 3) on a similar ration produced eggs of average low hatchability, appear to confirm the statement that this difference may be accounted for on the basis that a number of the hens in the previous ex-
TABLE 6.—Record of feed consumption and hen performance—21-40 weeks Lot No.
Distinguishing feature of rations used
Flavin per Feed Riboflavin Riboflavin Produc- gm. eggs, Hatch ol per hen per gm. per hen fertile tion air dried day day feed eggs micrograms
pet.
1
Basal ration
40
micrograms 0.80
2
5 percent dehydrated alfalfa leaf meal
89
1.50
133.5
34.5
10.0
35.6
10 percent dehydrated alfalfa leaf meal
94
1.90
178.6
51.0
20.0
62.8
3
micrograms 32.0
pet. 0.0
4
5 percent dried skimmilk
84
1.60
134.4
45.0
11.0
9.2
5
10 percent dried skimmilk
85
2.30
195.5
44.7*
17.0
48.1
78
2.20
171.6
48.0*
16.0
72.9
6
5 percent dried skimmilk+ 5 percent dehydrated alfalfa leaf meal
* High range paralysis in these lots.
correlation is not shown in those lots receiving 10 percent dried skimmilk or 5 percent each of alfalfa leaf meal and dried skimmilk in their ration. From previous experiments (part one, table 2) and from the riboflavin content of the ration, the feed per hen day (Table 6, lots 5 and 6) is lower than expected, indicating that an abnormal condition existed in these lots. The fact that range paralysis developed in these lots during this period not only may account for the low feed consumption and low egg production but may also have been an (inherent) inhibiting factor against the predictable effect on growth, based on the riboflavin consumption in the two previous periods as shown in Tables 4 and 5.
periment (Table 2) produced eggs whose embryos were inherently weak. This confirms the findings of Davis, Norris, and Heuser (1938) with reference to the variation in hatchability of eggs from different hens due to difference in inheritance. It is interesting to note that pullets reared on the basal ration did not produce any eggs; whereas pullets reared on a good ration and then transferred to the basal ration (part one) produced normally for a few weeks and then gradually declined. This indicates that the hen stores some riboflavin which is utilized during periods when riboflavin deficient rations are used. It is an interesting fact that the hatchability of eggs from hens receiving
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gm.
336
CHARLES H. HUNT, A. R. WINTER AND R. M. BETHKE
5 percent each of alfalfa leaf meal and dried skimmilk were consistently higher than the eggs from hens receiving 10 percent of dried skimmilk, both rations having about the same riboflavin content. This indicates that a combination of alfalfa leaf meal and dried skimmilk has some factor other than riboflavin which effects embryonic development. Whether these findings can be explained on the basis of the observations of Bauernfeind et al. (1938) that another unknown factor is involved in embryonic development, requires further study.
The amount of feed consumed per hen day increased as the amount of riboflavincarrying ingredients in the ration is increased. The riboflavin content of the egg varied directly with the riboflavin content of the ration of the hen. Eggs of the highest riboflavin content gave the highest hatchability and the chicks from such eggs showed the highest livability when fed a ration deficient in riboflavin. The hatchability results indicate that approximately 220-230 micrograms of riboflavin per 100 grams of feed are required for the production of good hatchable eggs. The data also suggest that the riboflavin requirements for/ good egg production are somewhat less than for maximum hatchability. / The needs Joi riboflavin for maximum growth in chicks to 12 weeks of age are about 190-2(30 micrograms per 100 grams of feed. /
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REFERENCES
Bauernfeind, J. C , A. E. Schumacher, A. Z. Hodson, L. C. Norris, and G. F. Heuser, 1938. A new factor required for growth and reproduction in the domestic fowl. Proc. Soc. Exp. Biol, and Med. 39:108-111. Bethke, R. M., P. R. Record, and D. C. Kennard, 1931. A type of nutritional leg paralysis affecting chicks. Poul. Sci. 10:355-368. , 1933. Relation of the vitamin G complex to hatchability and nutritive value of eggs. Poul. Sci. 12:332-333. , P. R. Record, and O. H. M. Wilder, 1937. Further studies on vitamin G in chick nutrition. Poul. Sci. 16:175-182. Davis, H. J., L. C. Norris, and G. F. Heuser, 1938 The role of vitamin G in reproduction in poultry. Poul. Sci. 17:81-86. , 1938. Further evidence on the amount of vitamin G required for reproduction in poultry. Poul. Sci. 17:87-93. Heiman, V., 1935. Relative vitamin G contents of dried whey and dried skimmilk. Poul. Sci. 14:137-146. Heuser, G. F., H. S. Wilgus, Jr., and L. C. Norris, 1938. The quantitative vitamin G requirements of chicks. Poul. Sci. 17:105-108. Hunt, Chas. H., P. R. Record, and R. M. Bethke, 1935. Effect of the stage of maturity and method of curing upon the vitamin B and vitamin G content of alfalfa, clover, and timothy hays. J. Agr. Res. 5:251-258. , 1936. Observation on the vitamin G requirement of the chicken. Ohio Agr. Exp. Sta. Bimonthly Bulletin 21:127-130. Lepkovsky, S., and T. H. Jukes, 1935. The vitamin G requirement of the chick. J. Biol. Chem. 111:119-131.
Lepkovsky, S., L. W. Taylor, T. H. Jukes, and H. J. Almquist, 1938. The effect of riboflavin and the filtrate factor on egg production and hatchability. Hilgardia 11:559-591. Norris, L. C , G. F. Heuser, and H. S. Wilgus, Jr., 1930. Is the chief value in milk for feeding poultry due to the presence of a new vitamin? Poul. Sci. 9:133. Norris, L. C , H. S. Wilgus, Jr., A. T. Ringrose, V. Heiman, and G. F. Heuser, 1936. The vitamin G requirement of poultry. Cornell Agr. v / E x p . Sta. Bulletin 660.
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SUMMARY AND CONCLUSION
/