Unidentified Factors in Turkey Nutrition Affecting Hatchability and Progeny Growth 1

Unidentified Factors in Turkey Nutrition Affecting Hatchability and Progeny Growth 1 S. P. TOUCHBUEN, V. D. CHAMBERLIN AND E. C. NABEE Department of Poultry Science, Ohio Agricultural Research and Development Center, Wooster, Ohio 44691 (Received for publication April 2, 1971) ABSTRACT A series of six experiments were conducted over a six-year period to determine the effect of rearing conditions, breeder housing conditions and diet composition, on hatchability of eggs and poults growth due to transfer of unknown dietary factors from the breeder hen diet. Response to the unidentified factor supplements was enhanced by confinement rearing of the poults, feeding of simplified growing rations and maintaining the turkeys on slatted floors or in wire cages during egg production. The response obtained from complete diet was shown to be due to fish meal and a dried fish solubles supplement. In most of the experiments the fish factor response was observed toward the end of the reproductive period indicating a depletion of the factor from the hen during egg production. The data suggest that the fish factor is transferred to the egg where some benefit on hatchability is usually evident and that the factor is present in the young poult where it influences early growth particularly when the poult diet is devoid of a fish factor supplement. INTRODUCTION FOLLOWING the availability in 1950 of vitamin B 12 in crystalline form, it was quickly discovered that unknown dietary factors existed in a variety of feedstuffs which were required for optimum performance of poultry. Olcese and Couch (1950) fed a sucrosesoybean protein diet adequate in all the known nutrients to Leghorn hens in cages with raised wire floors. The injection of vitamin B 12 into the eggs was ineffective in preventing a decline in hatchability which occurred after 8 weeks on the experimental diet. The early embryonic mortality was attributed to the depletion from the body of the hen of an unknown dietary essential. Peeler et al. (1951) reported a similar decrease in hatchability of eggs from Leghorn hens in cages fed a more practical ration of corn, wheat, soybean meal, and containing all the known nutrients, but the depletion required more than one year. Condensed fish solubles were shown to supply an unknown hatchability factor 'Ohio Agricultural Research and Development Center Journal Article No. 33-71. POULTRY SCIENCE 51: 96-103, 1972 for New Hampshire pullets (Arscott and Combs, 1953). These workers noted a greater response if the hens were maintained in pens with raised wire floors. Thus, coprophagy was suggested as influencing the amount of the factor required in the ration. A slight carryover of the unidentified factor in the ration of the dams was shown to partially meet the requirements of the chicks. Green feeds were also found to be an unidentified factor source. Jacobs et al. (1953) demonstrated that the addition of five percent of dehydrated alfalfa leaf meal to a corn-soybean meal ration improved both the fertility and the hatchability of eggs from Leghorn pullets housed in cages. Touchburn et al. (1957) obtained a slight but statistically significant increase in hatchability upon the addition to the ration of 2.5 or 5 percent of either dehydrated cereal grass or dehydrated alfalfa meal. These ingredients were added as supplements to a practical ration and fed to New Hampshire chickens maintained on a litter floor. 96 Turkeys fed an all vegetable breeder ration became depleted of a factor necessary

UNIDENTIFIED FACTORS 97 for early embryonic development (Feldman et al., 1957). The depletion was not apparent until after 11 weeks on the experimental ration. Dehydrated alfalfa meal, dried brewer's yeast, and condensed fish solubles added as individual supplements to the basal diet each improved hatchability. A combination of these supplements was effective in preventing the early embryonic mortality and maintaining a high level of hatchability for the entire 24 week period of the test. Waibel (1958) reported unidentified growth factor activity in fish meal, alfalfa meal, distillers dried solubles, dried whey, and grass juice for the growing poult. The responses observed in the late hatched poults were not apparent in poults hatched early in the reproductive season from the same breeding stock. The purpose of the present study was to study the factors affecting the response of turkeys to unidentified factors in the breeder diet. GENERAL EXPERIMENTAL PROCEDURE Simplified or semi-purified starter, grower, finisher, and breeder diets composed of corn, soybean meal, and pure sources of vitamins and minerals were developed for this study. They provided known nutrients, presumably in excess of the required amounts, but were devoid of ingredients commonly considered as sources of unidentified factors. Simplified corn-soybean diets were tested against the regular Ohio turkey diets (complete diets) which contained in addition, sources of animal protein and unidentified factors. In later tests purified diets based on glucose and isolated soybean protein were used. The composition of the complete and simplified breeder diets are shown in Table 1. The composition of the purified breeder diet is shown in Table 2. Starter and grower diets were formulated in the same manner as the breeder diets except that the amounts of the major protein TABLE 1. Composition of complete and simplified turkey breeder diets Major Ingredients: Complete Corn-Soy Ground Yellow Corn Pulverized Oats Soybean Meal (44 protein) DL-methionine Wheat Middlings (17 protein) Meat and Bone Scrap (50 protein) Menhaden Fish Meal (60 protein) Dried Whey Product (16 protein) Alfalfa Meal (17 protein) Dried Fish Solubles Defluor. Rock Phosphate (31 Ca; 18 P) Ground Limestone (38 Ca) Salt Minor Ingredients 56.0 10.0 10.0 2.5 2.5 67.7 25.0 0.05 5.0 1.5 3.0 5.0 0.5 100.0 Minor Ingredient Additives Per 45.4 Kg. 1.0 0.75 0.5 100.0 Vitamin A, U.S.P. units 200,000 500,000 Vitamin D-3, l.c. units 81,000 100,000 Riboflavin, mg. 210 400 Pantothenic Acid, mg. 350 1,000 Niacin, mg. 1,500 2,500 Choline Chloride, mg. 39,000 39,000 Vitamin B-12, mg. 0.6 0.8 Vitamin E, I. units 800 2,000 Vitamin K, mg. 200 Pyridoxine, mg. 200 Folic Acid, mg. 100 Manganese Sulfate 12.0 22.7 (27 Mn), Zinc Sulfate (36 Zn), 3.6 4.5 Antioxidant (B.H.T.), 5.7 Calculated A tier age Composition Protein, 16.61 Productive Energy, 1,944 Kcal./kg. Fiber,. 4.49 Calorie/Protein Ratio 117 17.55 2,034 2.81 116 and energy source were varied to produce suitable protein-energy ratios. Spring hatched female poults of a medium weight White variety were fed the complete or simplified starter, grower, and finisher diets. Then the experimental breeder diets were fed beginning one month before the reproductive period. Oyster shell was available in the breeder pens on a free choice basis. Hens were artifically insemi-

98 S. P. TOUCHBUEN, V. D. CHAMBERLIN AND E. C. NABER TABLE 2. Composition of purified turkey breeder diet Major Ingredients: Glucose (Cerelose) Isolated Soybean Protein (C-l Assay Protein) Ground Cellulose (Solka-floc) Corn Oil Mineral Mix Vitamin Mix DL-Methionine 67.42 19.50 1.85 2.50 8.56 0.02 0.15 100.00 Minor Ingredients (supplied by Mineral and Vitamin mix in units per kg. of diet) CaC0 3 34.0 gms. CaHP0 4-2H 2 0 29.0 gms. NaCl K 2 HP0 4 6.0 gms. 11.1 gms. M 9 SCv7H 2 0 S.Ogms. MnS0 4 102 mgs. Fe Citrate CuS0 4-5H 2 0 333 17 mgs. mgs. NaMo0 4-2H 2 0 0.8 mgs. nated with pooled semen from toms receiving the complete breeder diet. Records were obtained on daily egg production and eggs were set in the incubator weekly. Infertility and embryonic mortality were verified by breaking and grossly examining the eggs. To study the effect of the parental treatment on the growth of progeny, poults were sexed at day old, distributed into battery brooders and fed complete, corn-soybean, or glucose-isolated soybean protein starter diets. In-so-far as possible, each battery compartment contained one male and one female poult from each parent treatment group. The number of replications varied from 18 to 40, depending on the number of poults hatched. Poults were weighed at 2, 3, and 4 weeks of age. The data were subjected to a least squares analysis of variance (Steel and Torrie, 1960) which allowed for unequal poult numbers arising from differences in hatchability and mortality. In conjunction with each progeny Niacin Riboflavin Ca Pantothenate Menadione Cyanocobalamin Pyridoxine-HCl Folacin Thiamine-HC1 Biotin Retinol Acetate Cholecalciferol Alpha Tocopherol 55.0 mgs. 8.8 mgs. 22.0 mgs. 4.4 mgs. 22.0 megs. 3.5 mgs. 4.0 mgs. 10.0 mgs. 0.2 mgs. 10,000 I. U. 2,200 I. C. U. 44.0 mgs. Acetate Para Aminobenzoic Acid m Inositol 20.0 mgs. 1,000 mgs. Butylated Hydroxyanisole 2.5 mgs. TABLE 3. Effect of rearing condition and breeder diet on fertility and hatchability Rearing Condition Range reared Confinement reared Experiment 1 Complete Diet 75.7 73.2 79.8 72.1** Diet growth test, another test measured the growth promoting effect of the supplement when added directly to poult diets. Poults used in these tests were progeny of hens fed the simplified or purified basal diets. These tests were conducted with poults hatched from eggs laid at the beginning, in the middle, and at the end of the reproductive season. Fer- Hatchtility ability Fer- Hatchtility ability 75.1 67.6 71.8** 68.3** ** Highly significant reduction in hatchability (p<.01).

UNIDENTIFIED FACTORS 99 SPECIFIC EXPERIMENTAL PROCEDURES AND RESULTS During the first year (Experiment 1), half of the pullets were reared on range after 16 weeks of age, the other half in confinement in a pole barn. Equal numbers of each group received either the complete or the simplified diets. For the reproductive season all birds were housed in the pole barn in pens with wood shavings litter. Each rearing condition and ration was represented by 4 pens of 20 hens each. The experimental treatments had no effect on rate of egg production or body weight of the hens. The average fertility and hatchability for the twenty-week reproductive period are presented in Table 3. The highest hatchability was obtained by range rearing of birds and feeding the complete breeder diet. Confinement rearing or feeding the simplified corn-soybean meal diet caused highly significant depressions of hatchability and the lowest level of hatchability was obtained by confinement rearing of birds and feeding the corn-soybean diet. This combination also resulted in a lower level of fertility. These same treatments also tended to reduce growth rate to 4 weeks of age with the progeny hatched in the middle of the reproductive period, but the differences were not statistically significant. Too few poults hatched toward the end of the period to provide critical growth tests. TABLE 4. Effect of breeder diet and poult diet on four-week weights of poults Experiment 2 Poult Diet Complete Complete 437 421 459* 407** * Significant increase in weight (p<.05). ** Highly significant decrease in weight (p<.01). TABLE 5. -Effect of seasonal depletion of dams fed a simplified diet on 4 week weights of the poults Experiment 2 Poult Diet Complete 1 gms. 462 458 Progeny Growth Test 2 3 gms. gms. 484 448 478 414** ** Highly significant reduction during Test 3 (p<.01). For Experiment 2 all the pullets were reared in confinement. They received either the complete or simplified diets during the growing and reproductive periods. For the reproductive period half of the hens on each diet were kept on litter floors, the other half on raised wooden slat floors. Although the data are not presented in tabular form hatchability was significantly reduced by feeding the simplified diet confirming the results of Experiment 1. The slat floor treatment resulted in a slight decline in hatchability in latter part of the season. When poults hatched at the end of the reproductive season were fed either the complete or simplified starter diets there was a highly significant breeder diet X poult diet interaction on 4-week weights of poults (Table 4). That is, poults fed the corn-soybean diet grew less rapidly than those fed the complete diet but the differences reached a highly significant level only in those from parents fed the corn-soybean breeder diet. The breeder pen floor type had no consistent effect on progeny growth. In Table S the average weights of the poults clearly demonstrates the effect of seasonal depletion. The average weights of the poults fed the corn-soybean starter diet were not significantly lower than those fed the complete starter diet in the first and second progeny growth tests. These poults were hatched from eggs produced after the first and tenth weeks of production, respec-

100 S. P. TOUCHBURN, V. D. CHAMBERLIN AND E. C. NABER tively. Poults for the third progeny growth test were hatched from eggs produced after 18 weeks of production. Here, the effect of feeding the corn-soybean starter diet was a highly significant depression of 4-week weight compared to that on the complete diet. This is in agreement with the report of Feldman et al. (1957) that turkeys fed an all-vegetable breeder diet became depleted of a factor necessary for early embryonic development only after 11 weeks. In Experiment 3 the dietary and housing treatments giving significant results in the previous two years were tested simultaneously. All the hens were fed either the complete or the simplified diets throughout the growing and reproductive periods. After 12 weeks of age, half of each group was raised on grass range, the other half in confinement. For the reproductive period half of the above treatment groups were housed in pens with litter floors, the other half in pens with raised wooden slat floors. The average hatchability data for the twenty : week reproductive season are summarized in Table 6. Hatchability was slightly reduced by feeding the corn-soy- TAELE 6. Effect of rearing condition, breeder diet and floor type on hatchability Experiment 3 and Floor Type Complete Diet Eitter Slats Average Diet Litter Slats Average Rearing Condition Range 72.4 67.7 70.1 68.7 70.0 69.9 Confinement 69.8 69.2 69.5 66.0* 62.2** 64.1** * Significant reduction in hatchability (p<.05). ** Highly significant reduction in hatchability (p<.01). TABLE 7. Ejfect of rearing condition, diet, and floor type of breeders on four-week weights of poults fed corn-soy starter diet 1 Experiment 3 and Floor Type Complete Diet Litter Slats Average Diet Litter Slats Average Range 444 412 Rearing Condition 433 472 407 440 Confinement 446 438 442 436 400* 418 1 Poults hatched at end of reproductive season reared in battery brooders and fed the corn-soybean basal poult diet. * Significant reduction in weight (p<.05) for slat floor. Average for both floor types approaches significance. bean diet, by rearing in confinement, and by housing on raised wooden slats, but a combination of these treatments was necessary before a significant difference became apparent. In progeny growth tests the same trend was observed but was less consistent (Table 7). The most interesting response was the reduced weight of poults from hens reared in confinement and fed the corn-soybean breeder diet. These differences approached significance at the 5 level of probability. These results lend support to the findings of Experiments 1 and 2 that maximum performance is enhanced by range rearing, complete diets, and litterfloored breeder pens. Conversely, the greatest sensitivity to unidentified dietary factors would be expected with confinement rearing, simplified diets, and slat floored housing of the breeders. The pullets for Experiment 4 were fed the simplified diet and reared in confinement on raised wooden slat floors after 12 weeks of age. One month before the start of the breeding season the birds were placed

UNIDENTIFIED FACTORS 101 in individual laying cages in an insulated building with a forced air heating system. The temperature in this building was generally held at 16 Centigrade, but inadvertently exceeded this temperature on occasion. The average egg production and hatchability in this unit were significantly lower than those obtained with a control group remaining in the slat floored pen in the pole barn where temperatures approximated the cold winter conditions outside. Supplements to the corn-soybean basal diet included 2.5 of fish meal plus 1.5 of dried fish solubles, or 5 of dehydrated alfalfa meal. These levels were the same as those added to the complete breeder diet. The supplemented diets were kept isonitrogenous by adjustment of the level of corn and soybean meal. Methionine level was maintained by varying the amount of DL-methionine added. There were no significant differences among treatment groups in the cage unit. This was not surprising in view of the decreased level of egg production and hatchability resulting, presumably, from decreased feed consumption. For example, on the corn-soybean basal diet feed intake was reduced to 190 grams per hen per day in the cage unit compared to 237 grams per hen per day in the pole barn. In the subsequent year the temperature of the laying cage unit was regulated to about 0-3 Centrigrade at night and 7-10 Centigrade during the day. For Experiment 5 the hens were reared under the previously described depletion conditions prior to housing in the individual laying cages. The breeder diets fed were as follows: (1) complete, (2) corn-soybean, (3) corn-soybean plus 2.5 of fish meal, and 1.5 of dried fish solubles, (4) corn-soybean plus 2.5 of dried whey product, (5) corn-soybean plus fish plus whey and (6) corn-soybean plus fish plus whey plus 5 of dehydrated alfalfa meal. TABLE 8. Effect of breeder diet and housing on 4- iveek weights of progeny fed a corn-soybean starter diet Experiment 5 Breeder Housing n Pole Barn Complete 415 419 384** 417 +Fish 407 +Whey 384** +Fish+Whey 424 +Fish+Whey 426 422 +Alfalfa ** Highly significant reduction in weight (P<.01). A satisfactory level of feed intake, 235 grams per hen per day, was attained in the cages as a result of the cooler temperatures maintained. Egg production compared favorably, the average for 112 days being 52.1 eggs per hen in the cage unit versus 53.6 in the pole barn. The differences in egg production and hatchability were not significant but the progeny fed the corn-soybean starter diet showed striking differences in early growth (Table 8). Poults from the hens fed the corn-soybean diet or the corn-soybean plus whey diet grew significantly slower than those from hens fed the complete diet or the corn-soybean diet supplemented with fish or combinations of fish with whey or with whey plus alfalfa. Growth of progeny of hens maintained on litter floors in the pole barn was good and showed no differences due to the different breeder diets. In Experiment 6 hens were depleted and again housed in the laying cage unit. Two percent of dried fish solubles or four percent of dried whey product were fed as supplements to a corn-soybean diet and also to a glucose-isolated soybean-protein (purified) diet. The addition of dried fish solubles improved the hatchability of fertile eggs in both types of diet (Table 9), although the

102 S. P. TOUCHBURN, V. D. CHAMBERLIN AND E. C. NABER differences were statistically significant only in the case of the purified diet. Dried whey product, in contrast, showed no effect on improving hatchability. The addition of fish solubles to the purified breeder diet greatly increased the 4-week poult weight, especially when the poults were fed the corn-soybean starter diet (Table 10). The dried whey product, on the other hand, produced no carry-over on growth from the diet of the hen to the poult. These same samples of fish solubles and whey, however, were both shown to possess unidentified growth factor activity when added to the corn-soybean basal diet of depleted poults (Table 11). A combination of these supplements added to the corn-soybean diet resulted in a slightly greater response than either supplement alone. Growth of poults fed the purified basal diet was significantly less than for those fed the corn-soybean basal diet. Addition of dried whey product produced absolutely no response. In contrast, the response to dried fish solubles in the purified diet was even more pronounced than in the corn-soybean diet in spite of the fact that the actual weights were not as great as on the supplemented corn-soybean diets. DISCUSSION These experiments provide substantial evidence for the existence of an unidentified nutrient or nutrients in fish meal and TABLE 9. Effect Supplement of breeder diet on hatchability Experiment 6 Diet Hatchability Purified Diet None 49.3 33.3 Fish Solubles 55.2 44.1* Dried Whey Product 43.3 35.4 * Significant increase in hatchability (P <.05) on purified diet. TABLE 10. Effect of breeder diet on 4 week weights of poults Experiment 6 -)-Fish -f Whey Purified Purified+Fish Purified-)-Whey CS 505 492 490 428 524** 435 Poult Diet Purif. 411 429 427 399 433 398 ** Highly significant increase in weight (P<.01) on purified diet. dried fish solubles required by the turkey for maximum hatchability. The significant differences in fertility observed only in Experiment 1 probably represent the inability to distinguish infertility from very early embryonic mortality during this first year of the program. The data suggest that the same factor in the diet of the hen is transferred through the egg to the poult and results in increased early growth of the poult. Furthermore, the factor is active in promoting early growth when incorporated directly into the poults' diet. A depletion of the factor from the body of the hen during the course of the reproductive period was demonstrated. This depletion was enhanced by depriving the birds of access to grass range or litter and TABLE 11. Effect of poult diet on 4-week weights 1 Experiment 6 Supplement to Poult Diet 4-Week Weights Corn- Soybean Diet gms. Purified Diet (Glucose- Isolated Soybean) gms. None 436 384 Fish Solubles 496** 452** Dried Whey Product 488* 393 Fish Solubles+Dried 512** 454** Whey Product 1 Poults from parents fed corn-soybean diet. * Significant increase in weight (P<.05). ** Highly significant increase in weight (P<.01).

UNIDENTIFIED FACTORS 103 by feeding simplified corn-soybean meal or glucose-isolated protein diets. The response in improved hatchability and early growth of the progeny to the fish factor suggest that the unidentified factor response is not merely a response to a better balance of amino acids and that the response is not due to alterations in the microflora of the intestinal tract in the dam. An interesting observation concerns the responses to dried whey product. This material produced no activity whatsoever in hatchability or carryover effect from the dam to the poult. Yet the same sample of dried whey product, when incorporated in the diet of the poults, did elicit significant growth responses. The whey factor effect might therefore more logically be due to changes in the intestinal microflora of the poult. REFERENCES Arscott, G. H., and G. F. Combs, 1953. Further evidence for an unidentified hatchability factor in condensed fish solubles. Poultry Sci. 32: 730-733. DIAMOND AUTOMATION NOTES J. R. Brown has been appointed Sales Engineer, Diamond Automation, Inc., Farmington, Michigan. He will be responsible for the Company's line of automated and non-automated cage systems in North and South Dakota, Nebraska, Kansas, Minnesota, Iowa, Missouri, and Wisconsin. For the past five years, Brown has been District Manager with the Farm Equipment Division of Allis Chalmers. STERWIN NOTES Sterwin Laboratories, Inc., a subsidiary of Sterling Drug Inc. has appointed J. L. Jackson as a Sales Representative. He will cover Mississippi, southeast Louisiana and southern Alabama. A.H.I. NOTES NEWS AND NOTES (Continued from page 95) (Continued on page 116) Feldman, G. L., R. L. Atkinson, B. G. Creech, T. M. Ferguson, B. L. Reid and J. R Couch, 1957. The effect of dehydrated alfalfa meal, dried brewers yeast and condensed fish solubles on the reproductive performance of turkeys. Poultry Sci. 36: 792-797. Jacobs, R. L., J. F. Elam, J. H. Quisenberry and J. R. Couch, 1953. Dehydrated alfalfa leaf meal as a source of vitamins and unidentified factors fo: the mature fowl. Poultry Sci. 32: 812-816. Olcese, O., and J. R. Couch, 1950. Effect of injecting vitamin Bu into eggs from hens fed a diet low in vitamin B ]2. Poultry Sci. 29: 612-614. Peeler, H. T., R. F. Miller, C. W. Carlson, L. C. Norris and G. F. Heuser, 1951. Studies of the effect of vitamin B K on hatchability. Poultry Sci. 30: 11-17. Steel, R. G. D., and J. H. Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill Book Co., New York. Touchburn, S. P., J. Biely and B. March, 1957. The effect of dehydrated green feed on fertility and hatchability of eggs from three generations of New Hampshire chickens. Poultry Sci. 36: 591-595. Waibel, P. E., 1958. Effectiveness of unknown growth factors, antibiotic, and animal fat in turkey poult rations. Poultry Sci. 37: 1144-1149. Dr. James G. Affleck, eneral Manager, Agricultural Division, American Cyanamid Company, Princeton, New Jersey, was elected President of the Animal Health Institute for 1971-72. He succeeds Mitchell Chadrijan, Vice President and General Manager, Animal Health and Diagnostic Divisions, Schering Corporation, Bloomfield, New Jersey. I.A.P.I. NOTES Bruce H. Dutton has been appointed Programs Director of the Institute of American Poultry Industries. He has been on the staff of business magazines, including Product Design and Development, published by the Chilton Company. He was Sales Manager of Advertising Age, and Acting Publisher of Advertising and Sales Promotion magazine. He has also been involved in market research, sales forecasting, sales control systems, budgets, and acquisition activities. He was Manager of National