The Niacin Requirement of Young Chickens G. R. CHILDS,* C. W. CAEEICK AND S. M. HAUGE Departments of Poultry Husbandry and Agricultural Chemistry, Purdue University Agricultural Experiment Station, Lafayette, Indiana DANN and Handler (9) stated that the hatched chick contained ten times as much niacin as the unincubated egg. This led the authors to suggest that the hatched chick is able to synthesize niacin and is therefore independent of a dietary supply of this vitamin. Snell and Quarles (9) observed that niacin is synthesized by the chick embryo to the extent that the final amount present is twenty times that originally present in the unincubated egg. They presumed that the synthesizing powers of the embryo are retained by the chick and, if so, it would indicate that niacin is not required in the diet of the chick. Briggs et al. (9) reported that niacin is essential in the diet of chicks for optimal growth and to prevent "chick blacktongue." They found niacinamide equally effective in stimulating growth and preventing blacktongue. Studies by Briggs et al. (9) indicated that a dietary deficiency of niacin in chicks was characterized by suboptimal growth, blacktongue, decreased food consumption, marked lowering of the niacin and Coenzyme I content of the breast muscle, poor feather development and occasion- * Present address: McMillen Feed Mills, Decatur, Indiana. Journal Paper No. 57 of the Purdue University Agricultural Experiment Station. This investigation was supported in part by a grant from the Central Soya Company. (Received for publication September, 95) 55 ally either perosis or scaly dermatitis. They concluded that chicks receiving the particular basal ration used in the experiment were able to synthesize about onesixth of their total niacin requirement. Based on the work of Briggs et al. (9), the National Research Council (Cravens et al., 96) recommended.8 mg. of niacin per gms. of feed to meet the chick's requirement for this vitamin. Briggs (95) working with chicks, showed that tryptophan may replace niacin in a purified diet containing no corn. Also it was noted that gelatin in the ration greatly increased the requirement of the chick for niacin. Hunt et al. (97) conducted a study of the niacin content of nine double-cross corn hybrids grown at five experiment stations for three different years. The range of the average niacin content of the various corn hybrids was.69 mg. per gms. to.5 mg. per gms. Illinois 78 and U.S. were consistently low each year at each location, while Iowa 99 was consistently high. EXPERIMENTAL The chicks used in all trials were Barred Plymouth Rocks purchased from a nearby hatchery. All the chicks came from one breeding flock and were from hens fed a commercial breeding mash. The day-old chicks were sexed according to a method described by Gerry and Downloaded from http://ps.oxfordjournals.org/ at Penn State University (Paterno Lib) on September 6, 6
55 G. R. CHILDS, C. W. CARRICK AND S. M. HAUGE TABLE. Composition of the niacin-deficient diet Glucose (Cerelose) Commercial casein Gelatin Soybean oil DL-methionine (98% purity) Vitamin supplement Mineral supplement -A, -D oil Percent 6.. 8.... 6..5 Vitamin Supplement (mg. per gms. of feed): Choline CI, i-inositol, riboflavin.8, calcium pantothenate., pyridoxin HCl.6, alpha-tocopherol.5, thiamin HCl., folic acid., menadione ( me-naphthoquinone)., and biotin.. Mineral Supplement (gms. per gms. of feed): CaCOs.5, Ca (P )., K HP.85, NaCl-Iodized.87, Na HP.7, MgSCv7H.5, Fe(C 6 H 5 7 ) -6H., MnS -H.5, ZnS., CuSCv5H., H BO s.9, and CoCl-6H.. Mishler (99), wing banded, and all chicks were depleted for one week, when they were paired and distributed into their respective experimental pens. The composition of the niacin-deficient diet used for depletion is given in Table. The chicks were all reared in battery brooders with wire floors. Feed and tap water were supplied ad libitum during the experimental period. Individual weights were taken at bi-weekly intervals after pairing until the termination of the experiment. Chicks were starved at least five hours before being weighed. Individual gains were calculated for the experimental TABLE. Niacin content of ingredients for Trials - Ingredient U.S. Corn Expeller soybean meal Basal ration U.S. Corn Expeller soybean meal Gelatin Trials and and and and Meg. per gm.* 8.7 6.. 8.. Not measurable * Expressed as micrograms of niacin per gram of air dried material. period, from the date of pairing to the date of termination. These gains were analyzed statistically by the analysis of variance. The niacin content of the feed and liver samples was assayed microbiologically by the standard method outlined in "Methods of Vitamin Assay" (97) using Lactobacillus arabinosus 7-5 as the test organism. The results for the feed ingredients are shown in Table, and the results for liver assays are presented in the tables and discussions of the various trials. The tryptophan content of the various protein ingredients in Trial was deter- TABLE. Tryptophan content of ingredients for Trial Ingredients Percent tryptophan U.S. Corn.6 Soybean meal (expeller).87 Gelatin.6 * Expressed as percent of air dried material. mined microbiologically by the method of Dunn (95) using Lactobacillis arabinosus as the test organism. The results are shown in Table. The composition of the diet with respect to other vitamins and amino acids was estimated from published literature values and appropriate corrections made when necessary. RESULTS U. S. hybrid corn was used in all experiments, since it was relatively low in niacin content. Sufficient quantities of this corn and expeller soybean meal were obtained at one time and composited so that for any particular trial the corn and soybean meal samples were uniform throughout. Trial, conducted during April, May, and June, 95, was designed to deter- Downloaded from http://ps.oxfordjournals.org/ at Penn State University (Paterno Lib) on September 6, 6
NIACIN REQUIREMENT or CHICKS 55 mine the niacin requirement of growing chicks fed a diet consisting largely of corn and soybean meal. All chicks were fed the niacin-depletion diet for the first week (see Table ), paired within each sex on the basis of one-week body weights, and distributed uniformly into their experimental pens. Levels of niacin fed and the results obtained are given in Table. Examination of the -8 week gains (Table ) indicates a highly significant increase (com- Ingredients Basal Niacin (mg. %) added Niacin (mg. %) Total (which contained approximately.6 mg.) gave a significant increase in growth. The addition of.5 mg. of niacinamide to the basal diet gave some increase in growth but not enough to be significant. In all comparisons between equal amounts of niacin and niacinamide (compare s,, 5, and 7 with s,, 6, and 8 respectively), niacin consistently gave slightly better growth than niacinamide, but the differences were not significant. TABLE.- The quantitative requirements for niacin (Trial ) 5 () () ().6.6.6 5.6 6.6 6 (8).6 Av. paired gains,* -8 weeks (grams) 7 males, 7 females per lot: Males 89 9 896 87 85 856 868 Females 695 76 78 79 79 7 7 Both sexes 766 8 8 789 777 79 85 Mortality to 8 weeks Feed-gain ratio 5.7....6.7 7 (6) 8.6. One-week weights (grams): Males 6, females 55, both sexes 58. i Corn 56.95, soybean meal (expeller) 8, DL-methionine.5, choline chloride.5, riboflavin., calcium pantothenate.5, Lederle A.P.F..75, A and D feeding oil (,-A, -D).5, steamed bonemeal.5, calcium carbonate.5, iodized salt and MnSC>.5 percent. Approximate least significant differences: 5% level % level Males 56 grams 7 grams Females grams 56 grams Both sexes 5 grams 6 grams Sexes combined (-8 weeks). pare s and ) due to the addition of mg. of niacin to grams of the basal ration which already contained.6 mg. Still higher levels of niacin gave no added growth response; in fact, there seemed to be a tendency toward growth depression, but the differences were not consistent. Trial, conducted during June, July, and August, 95, was designed to test the relative efficacy of niacin and niacinamide. The rations fed and the results obtained are shown in Table 5. As in Trial, the addition of mg. of niacin to grams of the basal ration High levels of either niacin or niacinamide did not depress growth in this trial. Some perosis was observed in practically all the experimental lots. This condition was probably a result of the severe depletion technique, since it was rather evenly distributed throughout all levels of niacin and niacinamide. Trial was conducted during September, October, and November, 95, and was designed primarily to determine the relationship of tryptophan to the niacin requirement with a factorial experiment. Since it has been reported that tryptophan Downloaded from http://ps.oxfordjournals.org/ at Penn State University (Paterno Lib) on September 6, 6
55 G. R. CHILDS, C. W. CARRICK AND S. M. HATJGE TABLE 5. Comparative utilization of niacin and niacinamide (Trial ) Ingredients Basal' Niacin (mg. %) added Niacinamide (mg. %) added Niacin or Niacinamide (mg.%) Total ' 8.6 Av. paired gains,* -7 weeks (grams) males, 5 females per lot: Males 6 Females 555 Both sexes 59 Feed-gain ratio No. perosis Mortality to 7 weeks. 9 (.5).56 676 57 6.6 (.).6 7 568 66. (.).6 689 56 65.7 (.5).56 7 57 66.8 (.).6 75 597 65.6 (.).6 689 586 67. 5 5 (.) 5.6 687 58 6. 6 (.) 5.6 688 576 6. 7 (6.) * One-week weight (grams): Males 69, females 6, both sexes 66. Corn 56.95, soybean meal (expeller) 8, DL-methionine.5, choline chloride.5, riboflavin., calcium pantothenate.5, Lederle A.P.F..75, A and D feeding oil.5, steamed bonemeal.5, calcium carbonate.5, iodized salt and MnSOj.5 percent. Approximate least significant differences: 5% level % level Males 5 grams 8 grams Females 9 grams 65 grams Both sexes 9 grams 5 grams Sexes combined (-8 weeks). Basal DL-tryptophan Niacin (mg. %)- added added Ground yellow corn Protein % Tryptophan % -Total Niacin (mg. %)-Total Av. paired gains.,* -6 weeks (grams) 9 males, temales per lot: Males Females Both sexes Feed-gain ratio No. perosis Mortality to 6 weeks TABLE 6.- Relationship of 9 56.95 5..9.6 79..8 8. 56.9 5...6 76 98.7 6. 56.9 5.6..6 58 9.8 6 8.6 678 6 65.5 8 (6.) 8.6 687 598 6. can replace niacin in the diet of chicks fed a purified diet, it was deemed necessary to test this relationship when chicks were fed a corn-soybean meal diet. Components of the basal diet were assayed microbiologically for tryptophan (see Table ) and the basal diet was calculated to contain.9 percent tryptophan. Components of the basal diet were also assayed for niacin (see Table ) and values obtained showed.6 mg. of niacin per grams of the basal diet. Four levels of tryptophan (.9,.,., and.6 percent) were fed within each of three levels of niacin (.6,.6, and.6 mg.). Rations fed and results obtained are given in Table 6. On the low niacin level (.6 mg.), increasing tryptophan from.9 to.6 percent gave some additional growth, but tryptophan to the niacin requirement {Trial ).7 56.88 5.9.6.6 5 8 7.7 7 56.95 5..9.6 5 8 5.7. 56.9 5...6 5 6 96.7 5. 56.9 5.6..6 58 6 87.55 5 6.7 56.88 5.9.6.6 5 56 5.8 7 () 56.95 5..9.6 58 57 5. 8. () 56.9 5...6 55 8 58. 9. () 56.9 5.6..6 565 97 5..7 () 56.88 5.9.6.6 55 77 5. Downloaded from http://ps.oxfordjournals.org/ at Penn State University (Paterno Lib) on September 6, 6 * Two-week weights (grams): Males 6, females 6, both sexes 6. Soybean meal, gelatin 8, DL-methionine.5, choline chloride.5, calcium pantothenate.5, Lederle APF.75, A and D feeding oil.5, steamed bonemeal.5, calcium carbonate.5 and iodized salt and MnS.5 percent. Approximate least significant differences: 5% level % level Males 66 grams 87 grams Females 5 grams 7 grams Both sexes grams 56 grams Sexes combined (-6 weeks).
NIACIN REQUIREMENT OF CHICKS 555 the tryptophan, in amounts used in this trial, could not entirely compensate for a niacin deficiency. A diet high in tryptophan might be of value in correcting a "border-line" niacin deficiency, but otherwise the niacin replacing value of tryptophan was deemed to be of little practical importance in a corn-soybean meal ration. A comparison of the niacin-tryptophan interrelationship is given in Table 7. It should be pointed out that the average gains for niacin showed a consistent increase corresponding to increasing levels of niacin supplementation, but this is not true for the averages of the different levels of tryptophan. On the low niacin level (.6 mg.), an increase in the level of tryptophan did increase the gains somewhat, but there were no significant increases in the gains when additional tryptophan was added, when the diet contained an adequate level of niacin. Gains were greater on the.6 mg. level of niacin than on the.6 mg. level at all levels of tryptophan fed, but the differences were not consistently significant. Trial was conducted during January and February, 95. It was designed to determine to what extent the chicken was dependent upon a dietary source of niacin as it grew older and also to determine if the feeding of excess niacin enabled chicks to withstand a later niacin deficiency better than other chicks that had been fed on a level approximating the minimum requirement. All chicks were subjected to a one-week depletion period as described previously and s,,, and were fed for two weeks on a cornsoybean meal ration containing.8 mg. of niacin per grams of ration, while s 5, 6, 7, and 8 were fed for two weeks on a similar ration containing 5.8 mg. of niacin. Niacin was removed from the diets of s and 5 at three weeks of age, from s and 6 at five weeks of age, etc. The rations fed and the de- TABLE 7. Gains in grams, from -6 weeks, for a factorial set of niacin and tryptophan treatments Niacin mg. percent.6.6.6 Average Tryptophan, percent of diet.9 5 5 5. 98 96 58 67. 9 87 5 69.6 7 5 5 77 Average 86 96 56 tailed plan of experiment are given in Table 8. The ration fed the chicks after niacin was removed from the diet was a purified ration complete in all known nutrients, except that it had no measurable niacin. In another trial this ration, when supplemented with niacin, gave growth comparable to the complete corn-soybean meal ration. Liver samples were assayed for niacin. The samples were taken at the time the niacin was removed from the diet and again at the termination of the experiment, when the chicks were eight weeks of age. It was expected that the niacin content of the liver for various ages of chicks on the two levels of niacin supplementation might give a better indication of the requirement for niacin as the chicken grows older, but such was not the case. Results obtained from the various assays were not consistant, but two general trends were shown in most of the samples studied. They were: niacin tends to concentrate in the liver with increasing age regardless of the level of niacin in the diet, () under adverse conditions {e.g. removal of niacin from the diet) niacin tends to concentrate in the liver presumably at the expense of other body tissues. This interpretation, however, is not supported by definite evidence. The chicks were weighed at one week for pairing and then at biweekly periods Downloaded from http://ps.oxfordjournals.org/ at Penn State University (Paterno Lib) on September 6, 6
556 G. R. CHILDS, C. W. CARRICK AND S. M. HAUGE TABLE 8. The effect of removing niacin from the dtet of chicks at different ages (Trial ) Ingredients Basal* Niacin (mg. %) added Niacin mg. % Total Niacin removed from diet at.8 weeks.8 5 weeks.8 6 weeks.8 5 5.8 weeks 6 ()' 5.8 5 weeks 7 () 5.8 6 weeks Averaged paired gains, -8 weeks (grams) Males 595 75 877 79 69 766 95 Females 87 5 68 78. 6 89 6 69 Both sexes 5 558 68 8 59-685 8 Liver niacin at 8 weeks* 9 6 7 5 6 88 6 Ground yellow corn 56.75, soybean meal 8, DL-methionine.5, dry choline supplement (5% choline chloride).6, riboflavin., calcium pantothenate.5, Aurofae*.5, A and D feeding oil.5, steamed bonemeal.5, calcium carbonate.5, and iodized salt and MnS(V.5 percent. * Source of Bis and antibiotic. Approximate least significant differences: 5% level % level Mates 77 grams grams Females 9 grams 65 grams Both sexes grams 57 grams Expressed as micrograms of niacin per gram of fresh liver. 8 () until five weeks; after five weeks the chicks were weighed at weekly intervals until termination of the experiment at eight weeks. The -8 week gains are given in Table 8 and the gains for both sexes for the different periods are presented graphically in Figure.. Chicks fed on either.8 mg. or 5.8 mg. of niacin per grams of diet grew at about the same rate after niacin was removed from their diet. Therefore, it appears that the chick cannot store large amounts of niacin to meet the requirement for rapid growth, even though it is ingesting niacin in an amount much in excess of what is needed for optimum growth. which was fed.8 mg. of niacin per grams of diet for the entire period and 8 which was fed a diet containing 5.8 mg. of niacin for the entire period had almost identical gains. It was concluded from this comparison that.8 mg. percent of niacin is sufficient for optimum growth. DISCUSSION A corn-soybean meal diet containing as much as.6 mg. of niacin per grams gave greater growth responses when supplemented with additional niacin. The absolute minimum requirement for niacin was not determined in the series of completed trials, but growth data indicated that it was no lower than.6 mg. nor higher than.8 mg. These figures are not in agreement with the requirement as stated by Briggs et al. (9) but certain differences in the experimental conditions should be noted. At this Station, chicks on different treatments seldom show significant differences at four weeks of age. Finally, earlier studies were conducted with a purified diet, while our studies have been conducted with a "high-efficiency" corn-soybean meal diet, which gives rapid growth, when adequately supplemented with niacin. It was shown in Trial that the chicken is dependent upon a dietary source of niacin until at least 8 weeks of age. When niacin was removed from the diet of the various lots, a significant retardation of growth was evident in one or two weeks as compared to the lots that continued to receive a diet adequate in niacin. It is sometimes assumed that the chick's requirement for niacin decreases as it grows older. It was expected that data 5.8 Downloaded from http://ps.oxfordjournals.org/ at Penn State University (Paterno Lib) on September 6, 6
NIACIN REQUIREMENT OF CHICKS 557 8 GRAMS GAIN NIACIN IN DIET NO NIACIN INDIET 7 6 5.8MG,%NIACIN / I' ' / / / / / / / * / s / s / s / S /// * 5 6 / ' / / J / A o / * // s WEEKS 5. 8MG.% NIACIN / // V / / / / r * / ' S - / S' / / / / // y s /. / /s // '. i i '. 8 5 6 WEEKS FIG.. The effect on to 8 week gains of chicks when niacin was removed from the diet at various intervals. from the various liver assays would give evidence to support this assumption, but the analyses of the livers from the various lots in Trial did not show a consistent relationship to the amount of dietary niacin. From a study of the various assays it was concluded that the amount of niacin present in the chicken's liver is not a reliable estimate of the amount of niacin available to the various body tissues to aid in maintaining rapid growth. SUMMARY. With the corn-soybean meal diets used, the niacin requirement for rapid growth was adequately met with a dietary level of.8 mg. of niacin per grams of feed.. Niacin appeared to have a slight, but non-significant, supplementary advantage over niacinamide in a corn-soybean meal diet.. Excess tryptophan could spare niacin to a slight extent, but could not compensate entirely for a partial niacin deficiency.. The chicken is dependent upon a dietary supply of niacin until at least 8 weeks of age. 5. No consistent relationship was established between the dietary niacin level and liver storage of niacin. Liver storage 8 Downloaded from http://ps.oxfordjournals.org/ at Penn State University (Paterno Lib) on September 6, 6
558 NEWS AND NOTES of niacin is not a reliable estimate of the amount of niacin available to the chicken to support rapid growth. ACKNOWLEDGEMENTS We are indebted to the Dawe's Manufacturing Company for the dry vitamin A and D concentrates and choline chloride, to Lederle Laboratories for the APF supplement and Aurofac, to the Dow Chemical Company for the DL-tryptophan, and to the Borden Company for the casein and gelatin, used in these experiments. REFERENCES Briggs, G. M., Jr., R. C. Mills, C. A. Elvehjem and E. B. Hart, 9. Nicotinic acid in chick nutrition. Proc. Soc. Exp. Biol. Med. 5: 59-6. Briggs, G. M., Jr., T. D. Luckey, L. J. Tepley, C. A. Elvehjem and E. B. Hart, 9. Studies on nicotinic acid deficiency in the chick. J. Biol. Chem. 8:57-5. include an incubator room, egg storage rooms, poultry dressing room, offices, classrooms, and a research laboratory. UTAH NOTES Dr. J. Anderson has been appointed to the staff of the Poultry Department, Utah State College, Logan. He was born in Brigham City, graduating from Utah State College in 9. He obtained his doctor's degree from the University of Maryland in 95. He was employed by Merck and Company, New Jersey. The Utah Turkey Federation has made a grant of $, and the Utah Bankers' Association a grant of $,5 to the State College towards the study of staphylococcus arthritis or synovitis in turkeys. ILLINOIS NOTES Dr. W. A. Glista has been appointed an Assistant Director of Research for the Chas. M. Cox Co., Boston. He will head the nutritional activities at the Wirthmore Research Laboratory and Farm. He graduated from the University of Massachusetts NEWS AND NOTES {Continued from page 58) Briggs, G. M., Jr., 95. Influence of gelatin and tryptophan on the nicotinic acid requirement of chicks. J. Biol. Chem. 6: 79-75. Cravens, W. W., H. J. Almquist, L. C. Norris, R. M. Bethke and H. W. Titus, 9. (Revised 96). Recommended nutrient allowances for poultry. National Research Council. Dann, W. J., and P. Handler, 9. Synthesis of nicotinic acid by the chick embryo. J. Biol. Chem. :95-96. Dunn, M. S., M. W. Camien, S. Shankland and L. B. Rockland, 95. Procedures for the determination of amino acids by microbiological methods as employed in the chemistry laboratory. University of California, Los Angeles. Mimeograph. Gerry, R. W., and D. H. Mishler, 99. Sexing Barred Rock chicks. Poultry Sci. 8: 79-85. Hunt, C. H., L. Ditzler and R. M. Bethke, 97. Niacin and pantothenic acid content of corn hybrids. Cereal Chem. :55-6. Methods of Vitamin Assay, 97. Interscience Publishers, Inc., New York. Snell, E. E., and E. Quarles, 9. The effect of incubation on the vitamin content of eggs. J. Nutrition :8-89. in 97, majoring in poultry. The next year he obtained a master's degree at the University of Connecticut and last year completed the requirements for a doctor's degree at the University of Illinois. GEORGIA NOTES B. W. Hancock, who was supervisor of the Georgia National Egg Laying Test and Broiler Breeding and Testing Project at the University of Georgia, Athens, has resigned to join the Southern Poultry Farms, Winder, Ga., as geneticist. CALIFORNIA NOTES The new $5, state livestock and poultry pathology laboratory of the California Department of Agriculture was opened at Fresno on October, 95. The building, on a acre site, has, square feet of space. Dr. W. W. Worcester, formerly at the Turlock laboratory, is in charge. The first regional laboratory was completed in San Gabriel in January, 95, at a cost of $,. Work on a new $5, state poultry and livestock diagnostic laboratory at Petaluma will commence in February, 95. Downloaded from http://ps.oxfordjournals.org/ at Penn State University (Paterno Lib) on September 6, 6 {Continued on page 56)