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The Thiamine Requirement of Young Turkey Poults
The Thiamine Requirement of Young Turkey Poults RICHARD C.
ROBENALT*
Department of Poultry Husbandry, University of California, Davis (Received for publication July 6, 1959)
* Present address: School of Veterinary Medicine, University of California, Davis.
the dog and considerably less on a high-fat diet. It should be pointed out that in none of this work were purified diets used since their development did not come until later. Instead, these workers made use of natural feedstuffs which had been treated in various ways, and it is probable that some other factors were lost or destroyed. EXPERIMENTAL Broad Breasted Bronze poults were sexed at hatching and were segregated such that fhe weight range was 8 to 10 grams and averaged between 50 and 58 grams in all trials. This procedure eliminated weak and extra large birds which might have caused more variable results. In all trials the average weight of each of the pens was approximately the same and each group was composed of five males and five females. One pen was used for each experimental level of thiamine. The poults were kept in electrically heated battery brooders with raised wire floors. Feed and water were provided ad libitum and the birds were weighed twice weekly. In all but two trials the poults were placed on the experimental diet immediately. In the exceptions a commercial poult starter was fed for the first 24 hours. No appreciable difference was noted when this was done except that survival time was prolonged one extra day in the deficient pens. A satisfactory purified diet had to be devised which would meet all nutritional requirements of the poult except thiamine. The initial diet used a protein source of 20% acid-washed casein and 10% gelatin
354
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CONSIDERABLE amount of work has been done to establish the requirement for thiamine in the dog, rat, pig, and chick. Until now, no attempt has been made to establish the requirement for the turkey poult. The present work was undertaken to fill the gap in this area of knowledge. Light and Cracas (1938) reported a variation of 2 to 8 micrograms daily in three different strains of white rats, and Arnold and Elvehjem (1938a) found that growing rats needed 80 to 100 micrograms per 100 grams of ration for normal growth. Arnold and Elvehjem (1938b), using a ration composed of some natural feedstuffs which had been autoclaved, announced a requirement of 60 to 80 micrograms of crystalline thiamine chloride hydrochloride pen 100 grams of ration in the chick. In the growing pig, Hughes (1940) found a minimum daily requirement of 1 milligram of thiamine per 100 pounds of body weight, while Van Etten et al. (1940) reported a value of 106 to 120 micrograms per 100 grams of diet. In the development of a biological assay for thiamine by the use of chicks, Jukes and Heitman (1940) estimated the requirement for growth at 130 to 150 micrograms per 100 grams of diet under their conditions. They further reported that 100 micrograms would protect the chicks from deficiency symptoms. McCay (1943) cited a value of 100 micrograms per 100 grams of low-fat diet in
A
THIAMINE REQUIREMENT OF POULTS
added to the ration to balance the calciumphosphorus ratio. The new salt mix was fed at a level of 2%. When this new ration was tested there was considerable improvement in the growth and appearance of the poults but they were still far below that of birds on the commercial poult starter. On the assumption that the protein source might be inadequate, it was decided to change to a purified soybean protein known as Drackett Assay Protein C-l. This product when fed at a level of 30% plus 0.45% methionine, adequately meets the amino acid requirements of the poult. The mineral and vitamin content of Drackett protein is very low with thiamine only 0.25 micrograms per gram of dry product. Thus the protein contributed not more than 7 micrograms per 100 grams of ration. Some other minor changes were made to improve the ration. Vitamins A, D, and E were eliminated from the oil mix and included separately in a dry form. Cellu flour at 5% of the ration was substituted for an equal amount of the cerelose. This change was fairly successful in eliminating a pendulous crop condition which frequently occurs on highly purified diets. The final ration contained the following amounts per 100 grams of ration: Drackett protein C-l, 30.0 gm.; cerelose, 47.1 gm.; calcium gluconate, 5.0 gm.; cellu flour, 5.0 gm.; Ca 3 (P0 4 ) 2 , 4.0 gm.; soybean oil, 3.5 gm.; mineral mixture, 2.0 gm.; Na 2 HP0 4 -12 Hj.0,1.2 gm.; DL-methionine, 0.45 gm.; glycerol, 1.0 gm.; Dry A, 10,000 units/gm., 0.1 gm.; Dry D, 1500 ICU/gm., 0.1 gm.; d-alpha tocopherol acetate, 44 units/gm., 0.1 gm.; inositol, 0.1 gm.; P.A.B.A., 0.1 gm.; choline chloride, 0.25 gm.; calcium pantothenate, 3.0 mgm.; riboflavin, 1.0 mgm.; pyrodoxine, 1.0 mgm.; folic acid, 0.5 mgm.; biotin, 0.04 mgm.; vitamin Bi2, 1.0 micrograms: The special mineral mixture was made up of: sodium chloride, 250.0 gms.; manganese
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with added methionine. Mineral requirements were provided by a salt mix used in other studies by Kratzer et al. (1949). Instead of the usual vitamin mix which contains thiamine, all the known required vitamins were added individually to eliminate the possibility of deterioration from storage. A trial was run to test the experimental diet. Levels of 0, 75, 150 and 300 micrograms of thiamine per 100 grams of ration were fed plus a group on commercial poult starter. Results were highly erratic and not dependent on level of thiamine. The growth irregularities and sickly appearances of many of the birds led to the belief that the fault lay in destruction of one or more of the fat soluble vitamins. Thus the ration was modified by inclusion of an anti-oxidant, diphenyl-p-phenylenediamine (DPPD) and by adding the A and D feeding oil separately from the rest of the oil mix to avoid deterioration due to rancidity. When this new diet was tested the results were as erratic as before and many of the birds feathered poorly. At this point some work by Lyman and Elvehjem (1951) in rats and by Waibell et al. (1954) with chicks regarding the destruction of thiamine in purified diets was noted. Certain constituents of the salt mix were blamed. The most destructive ingredient was found to be K 2 HP0 4 , probably due to its great hygroscopicity and highly alkaline properties, but CaC0 3 and MnS0 4 •H 2 0 were also implicated. Waibell found thiamine relatively stable when the calcium, phosphorus, and potassium were supplied by Ca 3 (P0 4 ) 2 and KCl, and also that Na2HPC*4 did not destroy thiamine. Glycerol, at a level of 1%, replacing the sugar was found to protect the vitamin in a dry environment. Using this data, the salt mix was changed to eliminate K 2 HP0 4 , and CaC0 3 and CaH(P0 4 ) 2 were dropped from the ration. Ca 3 (P0 4 ) 2 and Na 2 HP0 4 • 12 H 2 0 were
355
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R . C. RoBENALT
pared and mixed into the rations by volumetric methods because of the relatively small quantities involved, and to assure adequate distribution. A commercial poult starter in the form of crumbles was fed to an extra pen in the earlier trials to serve as a positive control. This practice was discontinued when it became apparent that the diet was meeting growth requirements. RESULTS
sulfate, 10.0 gms.; sodium silicate, 100.0 gms.; ferric oxide, 10.0 gms.; copper sulfate, 4.0 gms.; zinc sulfate, 2.6 gms.; cobalt acetate, 1.0 gms.; potassium iodide, 0.6 gms.; aluminum sulfate, 12.6 gms.; magnesium sulfate, 200.0 gms.; potassium chloride, 400.0 gms. This ration was tested and compared with the casein-gelatin diet and with poult starter. Results with the casein-gelatin were similar to the previous ones while the new ration gave a growth curve similar to poult starter and equal on a feed conversion basis. Rations were freshly mixed just prior to the beginning of each trial. Thiamine chloride hydrochloride in 95% ethyl alcohol was added to the quantities of basal just prior to feeding. Alcohol was chosen as the solvent to prevent lumping of the cerelose in the ration. The thiamine solution was pre-
FIG. 2. Poult which received 50 micrograms of thiamine per 100 grams of diet. Note emaciation in head and legs and white runny feces. Age 12 days.
Poults which were fed the basal ration showed emaciation and progressive weakness resulting in inability to stand. Later the birds sprawled on their sides and finally died (Figs. 1 and 2). Little difference was noted in the reaction to a deficiency of thiamine by the poults and by chicks as reported by Jukes and Heitman (1940). The paralysis reported for chicks seldom appeared and only once was any head retraction noted. Runny white droppings were observed in deficient birds as can be seen in Fig. 2. The cause of these peculiar feces is not known. The growth curves obtained in comparable experiments were quite similar and the results of a typical experiment are illustrated by Fig. 3. A composite of the different trials was
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POULTS, IN DAYS
FIG. 3. Growth curve of poults fed various levels of thiamine.
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FIG. 1. Poult at left received 100 micrograms and one on right SO micrograms of thiamine per 100 grams of diet. Age 12 days.
THIAMINE REQUIREMENT OF POULTS
357
TABLE 1.—Body weight of poults on selected days fed various levels of thiamine Av. body wt. per bird in grams Thiamine Trial added per 100 gms. diet
Day 5
12
15
22
1
micrograms 0 100 200 300
SS 55 55 55
70 80 78 74
— 141 152 146
— 193 198 198
— 293 336 318
2
0 50 100 200 300
56 56 56 56 57
65 73 73 75 78
— — 120 142 130
— — 142 191 179
— — 205 304 328
0 100 200 300
57 57 57 57
62 75 76 73
— 121 111 119
— 156 146 151
— 176 246 262
0 50 100 200 300
59 57 57 56 57
58 67 76 80 82
— — 122 145 138
— — 159 181 171
— — 216 251 232
4
made and the average weights of poults at 12 days of age is shown in relation to the level of thiamine in the ration (Fig. 4). The data on selected days obtained in the four trials are shown in Table 1. The data upon which Fig. 4 is based are given in Table 2. The results indicate that approximately 100
150-
•
•
micrograms of thiamine per 100 grams of diet is required to meet minimum growth needs to 12 days of age. Two trials were made with S.C. White Leghorn chicks using the same procedures as for the poults. These experiments were conducted since no recent work had been done on the chick requirement using the purified diets and they would serve as a comparison of two related species under similar conditions. Table 3 gives the data on selected days for the two trials. A composite of the data from these trials is shown in Fig. S. The chick ration was similar to that for poults except for a reduction in the protein TABLE 2.—Composite of poult weight data at 12th day of age
)! O
\ 50
1
1 100
i 200
MICROGRAMS OF THIAMINE
I 300
PER 100 GRAMS OF RATION
FIG. 4. Thiamine level versus average poult weight on 12th day. At this age all poults receiving levels of 0 and 50 micrograms of thiamine had died.
Thiamine added per 100 gms. diet micrograms 0 50 100 200 300
Average body weight, grams Trial 1 — — 141 152 146
2 — — 120 142 130
Average 3
4
— — 121 111 119
— — 122 145 138
All dead All dead 125 138 133
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0
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R . C. ROBENALT
TABLE 3.—Body weight of chicks on selected days fed
various levels of thiamine Av body wt. per pen in grams Thiamine added Trial per 100 grams diet
V) ==200-
Day 0
5
12
15
— 115.4 129.0 131.6
22
micrograms 0 50 100 200 300
40.2 39.8 39.8 39.8 39.8
56.4 52.8 57.2 57.3 59.5
36.0 42.3 92.3 102.7 104.8
2
0 50 100 200 300
25.6 25.6 25.6 25.6 25.4
36.8 33.6 37.0 38.3 36.1
36.8 64.3 70.4 63.8
—
— — 85.9
93.1 85.6
—
— 189.4 205.8 205.1 — 140.9 152.2 142.1
level and in the calcium-phosphorus ratio to conform with the requirements of the chick. Thiamine was added at the same levels as in the poult experiments. The results obtained were similar to those with poults. The birds receiving no dietary thiamine were all dead in 11 to 12 days after being placed on the experimental diet while those on 50 micrograms survived only 4 to 5 days longer. All levels of 100 micrograms and over proved adequate, and the minimum requirement appears to be 100 micrograms. This agrees with the value previously reported by Jukes and Heitman (1940). DISCUSSION
Poults on the basal diet lost weight by the 5th day and were all dead by approximately the tenth day while those receiving 50 micrograms survived only about 2 extra days. All other levels of thiamine gave growth curves similar to poult starter and equal to each other at least until the twelfth day. At this time it was observed that the 100 microgram group began to grow more slowly. Instead of terminating one of the trials at the twenty-second day it was continued to 26 days and it was found that after the twenty-second day the 200 microgram group also showed a retardation in growth rate. Further discussion of this observation will be presented later. Since growth was about equal in all
groups receiving 100 micrograms or more in all trials up to the twelfth day of age it was apparent that this level was sufficient to protect the birds under the conditions studied. The data of the trials shown in Table 1 was statistically checked by means of the F-test for analysis of variance. The results were not significant at the 5% level for data up to the twelfth day. The original intention had been to establish the requirement up to the twenty-first day of age but the previously mentioned variations in the growth rate of the 100 microgram pen did not allow valid conclusions to be drawn from the results. Waibell et al. (1953) had reported that antibiotics showed an effect on chicks fed sub-optimum levels of thiamine so two further trials were conducted to see if this also held true in poults. The birds were fed levels similar to the previous trials except that a duplicate set of pens were given crude crystalline aureomycin, 90 to 92% pure, at a level of 25 milligrams per kilogram of ration. No effect of aureomycin on survival time was observed in the absence of thiamine although average weights of the surviving poults were somewhat greater. It was noted
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THIAMINE REQUIREMENT OF POULTS
When the results obtained in the poult and chick trials were compared with some other species a very close agreement was found. Rats, swine, and dogs were all reported to have a requirement very close to 100 micrograms. A notched beak condition frequently appeared during the different trials but it is believed to have no connection with the deficiency since it appeared in all levels of thiamine. It may possibly be related to a level of one of the non-essential amino acids or to some other minor factor not found in a purified diet but having no effect on growth. As has been previously mentioned, a slowing down of the rate of growth after a certain length of time was noted in pens on the lower levels of thiamine. It is possible that the requirement is actually higher than
has been reported and that the change is due to a depletion of body stores, or perhaps the requirement increases after 12 days of age. Another explanation is the possible destruction of thiamine in the ration. Waibell et al. (19S4) have reported a slow destruction of the vitamin by several of the components of the salt mixture besides the ones already implicated. These offenders could not be eliminated from the mix. One was MnS0 4 • H 2 0 which showed a delayed effect till after the 10th day. On this basis it would be expected that after a certain length of time the group receiving the 100 microgram level would slow down in rate of growth due to the reduction of thiamine to a sub-optimum level and that after another length of time the next higher level would also have enough thiamine destroyed to show a retardation. This indeed was observed in the one trial carried to 26 days. When the first chick trial was made, a similar change in growth rate was observed. Therefore, in the second chick trial an attempt was made to prevent this change by periodic mixing of fresh feed as needed. The full amount of basal was mixed but only l.S kilograms of diet was prepared for each group. After 13 days a second mix of the same size was made and 10 days later this was repeated. The result of this procedure was as expected, the growth rate of all pens remained the same. When poults were again available this procedure was tried with them. The final results obtained supported the expectations but extensive losses in the 100 microgram pen prevented valid conclusions from being drawn. At a later date the trial was repeated but with the addition of a second 100 microgram pen in which all feed was prepared at one time. However, accidental damage to this feed led to exclusion of this group from final results. Other figures were inconclusive except for percent gain per day
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that the retardation in rate of growth previously observed in the lower levels of thiamine was delayed several days in the presence of aureomycin while at higher levels there was no significant difference in the curves obtained. It seems apparent that antibiotics exert their effect only when thiamine is present at sub-optimum levels. In those poults receiving no dietary thiamine it is assumed there is some carry-over of the vitamin from the egg since during the first few days they show some gain and are able to survive to about 12 days. Also there is a small amount of thiamine present in the protein source as was previously mentioned but it is doubtful if this small amount is able to contribute much to the needs of the poult. Therefore, assuming there is some vitamin carried over from the egg, the antibiotic is acting in the presence of thiamine. The observation that the antibiotic increases the average weight but does not prolong the survival time is evidence that once the vitamin is entirely depleted there is no further action.
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in which all pens were approximately equal. Based on the results of these two trials it is not possible to state definitely that the theory regarding destruction of the thiamine by the mineral mix is correct. SUMMARY
NEWS AND NOTES (Continued from page 353) pointed out that, among his approximately 120 stu- of Poultry Science from 1921 to 1927, and was dents who are alumni of the Department during his President of the Association 1931 to 1932. He re44 years of active teaching, there are: an Experi- ceived the Resident Teaching Award of the Associament Station Director, three Directors of Research tion in 1950. From 1947 to 1952 he was Representative of the Poultry Science Association to the Poulfor commercial companies, sixteen Heads of Poultry Departments, twenty-two other Members of Poul- try and Egg National Board, and was General Chairman of the Program Committee for the Sevtry Department staffs, and the Research Workers for enth World's Poultry Congress in 1939. commercial companies. He was the co-author of over 80 scientific papers McMILLEN NOTES and bulletins, as well as many popular circulars and articles. Dr. D. F. Middendorf has been appointed He is a Fellow and Charter Member of the Poultry Research Specialist at the McMillen Feed Poultry Science Association, was Associate Editor Research Centre, Fort Wayne, Indiana. He received (Continued on page 389)
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Young turkey poults were fed varying levels of thiamine in a purified diet. Similar experiments were conducted with chicks. Poults deficient in thiamine show emaciation, runny white feces, progressive weakness causing inability to stand and finally death. The paralysis and head retraction found in chicks was seldom noted in poults. Under the conditions studied, 100 micrograms of thiamine per 100 grams of ration was adequate to prevent the appearance of deficiency symptoms and produce optimum growth in the young turkey poult to 12 days of age. This value was also sufficient for the chick under the same conditions. Growth of poults between 12 and 22 days of age was more rapid with 200 micrograms of thiamine per 100 grams of diet than with 100 micrograms. This may have been due to a slow destruction of thiamine, probably caused by some ingredients of the salt mix. This was prevented in one chick experiment by the periodic mixing of fresh feed.
REFERENCES Arnold, A., and C. A. Elvehjem, 1938a. Studies on the vitamin Bi requirements of growing rats. J. Nutrition, IS: 429-443. Arnold, A., and C. A. Elvehjem, 1938b. Studies on the vitamin Bi requirements of growing chicks. J. Nutrition, IS: 403-410. Hughes, E. H., 1940. The minimum requirement of thiamine for the growing pig. J. Nutrition, 20: 239-241. Jukes, T. H., and H. Heitman, Jr., 1940. Biological assay of thiamine with chicks. J. Nutrition, 19: 21-30. Kratzer, F. H., D. E. Williams and B. Marshall, 1949. The sulfur amino acid requirements of turkey poults. J. Nutrition, 37: 377-384. Light, R. F., and L. J. Cracas, 1938. Vitamin Bi requirements of different strains of white rats. Science, 87: 90. Lyman, R. L., and C. A. Elvehjem, 19S1. Lability of Bi in certain purified rations. Proc. Soc. Exp. Biol. Med. 77: 813-816. McCay, C. M., 1943. Nutrition of the Dog. Cornstock Publishing Co. Ithaca, New York. Van Etten, C , N. R. Ellis and L. L. Madsen, 1940. Studies on the thiamine requirement of young swine. J. Nutrition, 20: 607-622. Waibell, P. E., W. W. Cravens and C. A. Baumann, 1953. Effects of dietary antibiotics on the growth of chicks receiving suboptimum amounts of thiamine by mouth or by injection. J. Nutrition, SO: 441-450. Waibell, P. E., H. R. Bird and C. A. Baumann, 19S4. Effects of salts on the instability of thiamine in purified chick diets. J. Nutrition, 52: 273-283.
ROBENALT*
Department of Poultry Husbandry, University of California, Davis (Received for publication July 6, 1959)
* Present address: School of Veterinary Medicine, University of California, Davis.
the dog and considerably less on a high-fat diet. It should be pointed out that in none of this work were purified diets used since their development did not come until later. Instead, these workers made use of natural feedstuffs which had been treated in various ways, and it is probable that some other factors were lost or destroyed. EXPERIMENTAL Broad Breasted Bronze poults were sexed at hatching and were segregated such that fhe weight range was 8 to 10 grams and averaged between 50 and 58 grams in all trials. This procedure eliminated weak and extra large birds which might have caused more variable results. In all trials the average weight of each of the pens was approximately the same and each group was composed of five males and five females. One pen was used for each experimental level of thiamine. The poults were kept in electrically heated battery brooders with raised wire floors. Feed and water were provided ad libitum and the birds were weighed twice weekly. In all but two trials the poults were placed on the experimental diet immediately. In the exceptions a commercial poult starter was fed for the first 24 hours. No appreciable difference was noted when this was done except that survival time was prolonged one extra day in the deficient pens. A satisfactory purified diet had to be devised which would meet all nutritional requirements of the poult except thiamine. The initial diet used a protein source of 20% acid-washed casein and 10% gelatin
354
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CONSIDERABLE amount of work has been done to establish the requirement for thiamine in the dog, rat, pig, and chick. Until now, no attempt has been made to establish the requirement for the turkey poult. The present work was undertaken to fill the gap in this area of knowledge. Light and Cracas (1938) reported a variation of 2 to 8 micrograms daily in three different strains of white rats, and Arnold and Elvehjem (1938a) found that growing rats needed 80 to 100 micrograms per 100 grams of ration for normal growth. Arnold and Elvehjem (1938b), using a ration composed of some natural feedstuffs which had been autoclaved, announced a requirement of 60 to 80 micrograms of crystalline thiamine chloride hydrochloride pen 100 grams of ration in the chick. In the growing pig, Hughes (1940) found a minimum daily requirement of 1 milligram of thiamine per 100 pounds of body weight, while Van Etten et al. (1940) reported a value of 106 to 120 micrograms per 100 grams of diet. In the development of a biological assay for thiamine by the use of chicks, Jukes and Heitman (1940) estimated the requirement for growth at 130 to 150 micrograms per 100 grams of diet under their conditions. They further reported that 100 micrograms would protect the chicks from deficiency symptoms. McCay (1943) cited a value of 100 micrograms per 100 grams of low-fat diet in
A
THIAMINE REQUIREMENT OF POULTS
added to the ration to balance the calciumphosphorus ratio. The new salt mix was fed at a level of 2%. When this new ration was tested there was considerable improvement in the growth and appearance of the poults but they were still far below that of birds on the commercial poult starter. On the assumption that the protein source might be inadequate, it was decided to change to a purified soybean protein known as Drackett Assay Protein C-l. This product when fed at a level of 30% plus 0.45% methionine, adequately meets the amino acid requirements of the poult. The mineral and vitamin content of Drackett protein is very low with thiamine only 0.25 micrograms per gram of dry product. Thus the protein contributed not more than 7 micrograms per 100 grams of ration. Some other minor changes were made to improve the ration. Vitamins A, D, and E were eliminated from the oil mix and included separately in a dry form. Cellu flour at 5% of the ration was substituted for an equal amount of the cerelose. This change was fairly successful in eliminating a pendulous crop condition which frequently occurs on highly purified diets. The final ration contained the following amounts per 100 grams of ration: Drackett protein C-l, 30.0 gm.; cerelose, 47.1 gm.; calcium gluconate, 5.0 gm.; cellu flour, 5.0 gm.; Ca 3 (P0 4 ) 2 , 4.0 gm.; soybean oil, 3.5 gm.; mineral mixture, 2.0 gm.; Na 2 HP0 4 -12 Hj.0,1.2 gm.; DL-methionine, 0.45 gm.; glycerol, 1.0 gm.; Dry A, 10,000 units/gm., 0.1 gm.; Dry D, 1500 ICU/gm., 0.1 gm.; d-alpha tocopherol acetate, 44 units/gm., 0.1 gm.; inositol, 0.1 gm.; P.A.B.A., 0.1 gm.; choline chloride, 0.25 gm.; calcium pantothenate, 3.0 mgm.; riboflavin, 1.0 mgm.; pyrodoxine, 1.0 mgm.; folic acid, 0.5 mgm.; biotin, 0.04 mgm.; vitamin Bi2, 1.0 micrograms: The special mineral mixture was made up of: sodium chloride, 250.0 gms.; manganese
Downloaded from http://ps.oxfordjournals.org/ at Serials Section, Dixson Library on June 5, 2015
with added methionine. Mineral requirements were provided by a salt mix used in other studies by Kratzer et al. (1949). Instead of the usual vitamin mix which contains thiamine, all the known required vitamins were added individually to eliminate the possibility of deterioration from storage. A trial was run to test the experimental diet. Levels of 0, 75, 150 and 300 micrograms of thiamine per 100 grams of ration were fed plus a group on commercial poult starter. Results were highly erratic and not dependent on level of thiamine. The growth irregularities and sickly appearances of many of the birds led to the belief that the fault lay in destruction of one or more of the fat soluble vitamins. Thus the ration was modified by inclusion of an anti-oxidant, diphenyl-p-phenylenediamine (DPPD) and by adding the A and D feeding oil separately from the rest of the oil mix to avoid deterioration due to rancidity. When this new diet was tested the results were as erratic as before and many of the birds feathered poorly. At this point some work by Lyman and Elvehjem (1951) in rats and by Waibell et al. (1954) with chicks regarding the destruction of thiamine in purified diets was noted. Certain constituents of the salt mix were blamed. The most destructive ingredient was found to be K 2 HP0 4 , probably due to its great hygroscopicity and highly alkaline properties, but CaC0 3 and MnS0 4 •H 2 0 were also implicated. Waibell found thiamine relatively stable when the calcium, phosphorus, and potassium were supplied by Ca 3 (P0 4 ) 2 and KCl, and also that Na2HPC*4 did not destroy thiamine. Glycerol, at a level of 1%, replacing the sugar was found to protect the vitamin in a dry environment. Using this data, the salt mix was changed to eliminate K 2 HP0 4 , and CaC0 3 and CaH(P0 4 ) 2 were dropped from the ration. Ca 3 (P0 4 ) 2 and Na 2 HP0 4 • 12 H 2 0 were
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pared and mixed into the rations by volumetric methods because of the relatively small quantities involved, and to assure adequate distribution. A commercial poult starter in the form of crumbles was fed to an extra pen in the earlier trials to serve as a positive control. This practice was discontinued when it became apparent that the diet was meeting growth requirements. RESULTS
sulfate, 10.0 gms.; sodium silicate, 100.0 gms.; ferric oxide, 10.0 gms.; copper sulfate, 4.0 gms.; zinc sulfate, 2.6 gms.; cobalt acetate, 1.0 gms.; potassium iodide, 0.6 gms.; aluminum sulfate, 12.6 gms.; magnesium sulfate, 200.0 gms.; potassium chloride, 400.0 gms. This ration was tested and compared with the casein-gelatin diet and with poult starter. Results with the casein-gelatin were similar to the previous ones while the new ration gave a growth curve similar to poult starter and equal on a feed conversion basis. Rations were freshly mixed just prior to the beginning of each trial. Thiamine chloride hydrochloride in 95% ethyl alcohol was added to the quantities of basal just prior to feeding. Alcohol was chosen as the solvent to prevent lumping of the cerelose in the ration. The thiamine solution was pre-
FIG. 2. Poult which received 50 micrograms of thiamine per 100 grams of diet. Note emaciation in head and legs and white runny feces. Age 12 days.
Poults which were fed the basal ration showed emaciation and progressive weakness resulting in inability to stand. Later the birds sprawled on their sides and finally died (Figs. 1 and 2). Little difference was noted in the reaction to a deficiency of thiamine by the poults and by chicks as reported by Jukes and Heitman (1940). The paralysis reported for chicks seldom appeared and only once was any head retraction noted. Runny white droppings were observed in deficient birds as can be seen in Fig. 2. The cause of these peculiar feces is not known. The growth curves obtained in comparable experiments were quite similar and the results of a typical experiment are illustrated by Fig. 3. A composite of the different trials was
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POULTS, IN DAYS
FIG. 3. Growth curve of poults fed various levels of thiamine.
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FIG. 1. Poult at left received 100 micrograms and one on right SO micrograms of thiamine per 100 grams of diet. Age 12 days.
THIAMINE REQUIREMENT OF POULTS
357
TABLE 1.—Body weight of poults on selected days fed various levels of thiamine Av. body wt. per bird in grams Thiamine Trial added per 100 gms. diet
Day 5
12
15
22
1
micrograms 0 100 200 300
SS 55 55 55
70 80 78 74
— 141 152 146
— 193 198 198
— 293 336 318
2
0 50 100 200 300
56 56 56 56 57
65 73 73 75 78
— — 120 142 130
— — 142 191 179
— — 205 304 328
0 100 200 300
57 57 57 57
62 75 76 73
— 121 111 119
— 156 146 151
— 176 246 262
0 50 100 200 300
59 57 57 56 57
58 67 76 80 82
— — 122 145 138
— — 159 181 171
— — 216 251 232
4
made and the average weights of poults at 12 days of age is shown in relation to the level of thiamine in the ration (Fig. 4). The data on selected days obtained in the four trials are shown in Table 1. The data upon which Fig. 4 is based are given in Table 2. The results indicate that approximately 100
150-
•
•
micrograms of thiamine per 100 grams of diet is required to meet minimum growth needs to 12 days of age. Two trials were made with S.C. White Leghorn chicks using the same procedures as for the poults. These experiments were conducted since no recent work had been done on the chick requirement using the purified diets and they would serve as a comparison of two related species under similar conditions. Table 3 gives the data on selected days for the two trials. A composite of the data from these trials is shown in Fig. S. The chick ration was similar to that for poults except for a reduction in the protein TABLE 2.—Composite of poult weight data at 12th day of age
)! O
\ 50
1
1 100
i 200
MICROGRAMS OF THIAMINE
I 300
PER 100 GRAMS OF RATION
FIG. 4. Thiamine level versus average poult weight on 12th day. At this age all poults receiving levels of 0 and 50 micrograms of thiamine had died.
Thiamine added per 100 gms. diet micrograms 0 50 100 200 300
Average body weight, grams Trial 1 — — 141 152 146
2 — — 120 142 130
Average 3
4
— — 121 111 119
— — 122 145 138
All dead All dead 125 138 133
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0
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TABLE 3.—Body weight of chicks on selected days fed
various levels of thiamine Av body wt. per pen in grams Thiamine added Trial per 100 grams diet
V) ==200-
Day 0
5
12
15
— 115.4 129.0 131.6
22
micrograms 0 50 100 200 300
40.2 39.8 39.8 39.8 39.8
56.4 52.8 57.2 57.3 59.5
36.0 42.3 92.3 102.7 104.8
2
0 50 100 200 300
25.6 25.6 25.6 25.6 25.4
36.8 33.6 37.0 38.3 36.1
36.8 64.3 70.4 63.8
—
— — 85.9
93.1 85.6
—
— 189.4 205.8 205.1 — 140.9 152.2 142.1
level and in the calcium-phosphorus ratio to conform with the requirements of the chick. Thiamine was added at the same levels as in the poult experiments. The results obtained were similar to those with poults. The birds receiving no dietary thiamine were all dead in 11 to 12 days after being placed on the experimental diet while those on 50 micrograms survived only 4 to 5 days longer. All levels of 100 micrograms and over proved adequate, and the minimum requirement appears to be 100 micrograms. This agrees with the value previously reported by Jukes and Heitman (1940). DISCUSSION
Poults on the basal diet lost weight by the 5th day and were all dead by approximately the tenth day while those receiving 50 micrograms survived only about 2 extra days. All other levels of thiamine gave growth curves similar to poult starter and equal to each other at least until the twelfth day. At this time it was observed that the 100 microgram group began to grow more slowly. Instead of terminating one of the trials at the twenty-second day it was continued to 26 days and it was found that after the twenty-second day the 200 microgram group also showed a retardation in growth rate. Further discussion of this observation will be presented later. Since growth was about equal in all
groups receiving 100 micrograms or more in all trials up to the twelfth day of age it was apparent that this level was sufficient to protect the birds under the conditions studied. The data of the trials shown in Table 1 was statistically checked by means of the F-test for analysis of variance. The results were not significant at the 5% level for data up to the twelfth day. The original intention had been to establish the requirement up to the twenty-first day of age but the previously mentioned variations in the growth rate of the 100 microgram pen did not allow valid conclusions to be drawn from the results. Waibell et al. (1953) had reported that antibiotics showed an effect on chicks fed sub-optimum levels of thiamine so two further trials were conducted to see if this also held true in poults. The birds were fed levels similar to the previous trials except that a duplicate set of pens were given crude crystalline aureomycin, 90 to 92% pure, at a level of 25 milligrams per kilogram of ration. No effect of aureomycin on survival time was observed in the absence of thiamine although average weights of the surviving poults were somewhat greater. It was noted
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1
THIAMINE REQUIREMENT OF POULTS
When the results obtained in the poult and chick trials were compared with some other species a very close agreement was found. Rats, swine, and dogs were all reported to have a requirement very close to 100 micrograms. A notched beak condition frequently appeared during the different trials but it is believed to have no connection with the deficiency since it appeared in all levels of thiamine. It may possibly be related to a level of one of the non-essential amino acids or to some other minor factor not found in a purified diet but having no effect on growth. As has been previously mentioned, a slowing down of the rate of growth after a certain length of time was noted in pens on the lower levels of thiamine. It is possible that the requirement is actually higher than
has been reported and that the change is due to a depletion of body stores, or perhaps the requirement increases after 12 days of age. Another explanation is the possible destruction of thiamine in the ration. Waibell et al. (19S4) have reported a slow destruction of the vitamin by several of the components of the salt mixture besides the ones already implicated. These offenders could not be eliminated from the mix. One was MnS0 4 • H 2 0 which showed a delayed effect till after the 10th day. On this basis it would be expected that after a certain length of time the group receiving the 100 microgram level would slow down in rate of growth due to the reduction of thiamine to a sub-optimum level and that after another length of time the next higher level would also have enough thiamine destroyed to show a retardation. This indeed was observed in the one trial carried to 26 days. When the first chick trial was made, a similar change in growth rate was observed. Therefore, in the second chick trial an attempt was made to prevent this change by periodic mixing of fresh feed as needed. The full amount of basal was mixed but only l.S kilograms of diet was prepared for each group. After 13 days a second mix of the same size was made and 10 days later this was repeated. The result of this procedure was as expected, the growth rate of all pens remained the same. When poults were again available this procedure was tried with them. The final results obtained supported the expectations but extensive losses in the 100 microgram pen prevented valid conclusions from being drawn. At a later date the trial was repeated but with the addition of a second 100 microgram pen in which all feed was prepared at one time. However, accidental damage to this feed led to exclusion of this group from final results. Other figures were inconclusive except for percent gain per day
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that the retardation in rate of growth previously observed in the lower levels of thiamine was delayed several days in the presence of aureomycin while at higher levels there was no significant difference in the curves obtained. It seems apparent that antibiotics exert their effect only when thiamine is present at sub-optimum levels. In those poults receiving no dietary thiamine it is assumed there is some carry-over of the vitamin from the egg since during the first few days they show some gain and are able to survive to about 12 days. Also there is a small amount of thiamine present in the protein source as was previously mentioned but it is doubtful if this small amount is able to contribute much to the needs of the poult. Therefore, assuming there is some vitamin carried over from the egg, the antibiotic is acting in the presence of thiamine. The observation that the antibiotic increases the average weight but does not prolong the survival time is evidence that once the vitamin is entirely depleted there is no further action.
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in which all pens were approximately equal. Based on the results of these two trials it is not possible to state definitely that the theory regarding destruction of the thiamine by the mineral mix is correct. SUMMARY
NEWS AND NOTES (Continued from page 353) pointed out that, among his approximately 120 stu- of Poultry Science from 1921 to 1927, and was dents who are alumni of the Department during his President of the Association 1931 to 1932. He re44 years of active teaching, there are: an Experi- ceived the Resident Teaching Award of the Associament Station Director, three Directors of Research tion in 1950. From 1947 to 1952 he was Representative of the Poultry Science Association to the Poulfor commercial companies, sixteen Heads of Poultry Departments, twenty-two other Members of Poul- try and Egg National Board, and was General Chairman of the Program Committee for the Sevtry Department staffs, and the Research Workers for enth World's Poultry Congress in 1939. commercial companies. He was the co-author of over 80 scientific papers McMILLEN NOTES and bulletins, as well as many popular circulars and articles. Dr. D. F. Middendorf has been appointed He is a Fellow and Charter Member of the Poultry Research Specialist at the McMillen Feed Poultry Science Association, was Associate Editor Research Centre, Fort Wayne, Indiana. He received (Continued on page 389)
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Young turkey poults were fed varying levels of thiamine in a purified diet. Similar experiments were conducted with chicks. Poults deficient in thiamine show emaciation, runny white feces, progressive weakness causing inability to stand and finally death. The paralysis and head retraction found in chicks was seldom noted in poults. Under the conditions studied, 100 micrograms of thiamine per 100 grams of ration was adequate to prevent the appearance of deficiency symptoms and produce optimum growth in the young turkey poult to 12 days of age. This value was also sufficient for the chick under the same conditions. Growth of poults between 12 and 22 days of age was more rapid with 200 micrograms of thiamine per 100 grams of diet than with 100 micrograms. This may have been due to a slow destruction of thiamine, probably caused by some ingredients of the salt mix. This was prevented in one chick experiment by the periodic mixing of fresh feed.
REFERENCES Arnold, A., and C. A. Elvehjem, 1938a. Studies on the vitamin Bi requirements of growing rats. J. Nutrition, IS: 429-443. Arnold, A., and C. A. Elvehjem, 1938b. Studies on the vitamin Bi requirements of growing chicks. J. Nutrition, IS: 403-410. Hughes, E. H., 1940. The minimum requirement of thiamine for the growing pig. J. Nutrition, 20: 239-241. Jukes, T. H., and H. Heitman, Jr., 1940. Biological assay of thiamine with chicks. J. Nutrition, 19: 21-30. Kratzer, F. H., D. E. Williams and B. Marshall, 1949. The sulfur amino acid requirements of turkey poults. J. Nutrition, 37: 377-384. Light, R. F., and L. J. Cracas, 1938. Vitamin Bi requirements of different strains of white rats. Science, 87: 90. Lyman, R. L., and C. A. Elvehjem, 19S1. Lability of Bi in certain purified rations. Proc. Soc. Exp. Biol. Med. 77: 813-816. McCay, C. M., 1943. Nutrition of the Dog. Cornstock Publishing Co. Ithaca, New York. Van Etten, C , N. R. Ellis and L. L. Madsen, 1940. Studies on the thiamine requirement of young swine. J. Nutrition, 20: 607-622. Waibell, P. E., W. W. Cravens and C. A. Baumann, 1953. Effects of dietary antibiotics on the growth of chicks receiving suboptimum amounts of thiamine by mouth or by injection. J. Nutrition, SO: 441-450. Waibell, P. E., H. R. Bird and C. A. Baumann, 19S4. Effects of salts on the instability of thiamine in purified chick diets. J. Nutrition, 52: 273-283.