14494441 ma: 341w» 9/4 the Effect cf Unidentified tactcr Sources, Hntihictics and lt-a-tcccphernl Hcetate cn the Reproductive Performance ct Bread Breasted Brnnze and Beltsville Small tlthite turlreus TEXAS AGRICULTURAL EXPERIMENT STATION R. D. LEWIS. DIRECTOR, COLLEGE STATION._TEXAS SUMMARY This bulletin presents a review of the ‘work done in nutrition of turkey breeder hens at the Texas Agricultural Experiment Station since 1950. These investigations include tests with Broad Breasted Bronze and Beltsville Small White turkeys reared to maturity on the range or on wire floors. Various combinations, of unknown hatchability fac- tor sources. vitamins. antibiotics and antioxidants have been fed in an attempt to improve the repro- ductive performance of the turkey breeder hen. Increased hatchability of fertile eggs was ob- tained by the addition of unidentified hatchability sources (fish meal. liver distillers dried solubles. condensed fish solubles, dehydrated alfalfa meal. dried whey and dried brewers yeast) at levels of 3 to 5 percent to the diets of Broad Breasted ‘Bronze and Beltsville Small White turkey hens. The highest hatchability values were obtained when combinations of two or three of the above sources of unidentified factors were included as supplements in the breeder diet. Such combinations maintained hatchability at high levels for as long as 17 to 24 weeks. while the hatchability without these supplements dropped to extremely low levels after‘ the first 8 to 12 weeks of the laying period. The addition of supplemental vitamin E (D- alpha-tocopheryl acetate) to turkey breeder rations. in the presence of unidentified factor sources. pro- duced an additional increase in the hatchability of eggs. Unidentified hatchability factor sources included in the diet resulted in a decreased embryonic mor- CONTENTS A Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Introduction . .- . . . . . ., . . . . . . . . . . . . . . . . . . . . . . . Procedure . , . . . . . . . . . . _ . . . . . . . . . . . . f . . . . . . . . . . . . . . . Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Literature Cited . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . diet. without supplemental vitamin E. had tality during the first and last week of the '4 period. l’ The amount of tocopherol per egg y‘ preted as a measure of” vitamin E in t J reported for several experiments. The »-_ cate that unidentified hatchability factors Y oxidants included in the diet do not tocopherol content of the yolk. The vitamin E (in a stable form of D-alpha-t acetate) improved the hatchability of hens fed an all-vegetable protein diet. unidentified factors. These data indicate A ural feed ingredients may not contain vit a sufficient amount or in a form which is for use by the turkey hen. ‘- Embryos from hens fed an all-vegetab and bulging eyes. and a high percent embryos died during the last week of the ~- period. The addition of vitamin E to ~- hen diet reduced this embryonic mortality ~ vented the blindness. ' u The addition of dried brewers -»:~ meal and fish solubles. singly and in vari, binations. as supplements to an all-veget tein diet resulted in increased egg prod two experiments. The average amount required to produce an egg ranged from; 1.66 pounds. The combination of dried ._ yeast with condensed fish solubles prod v best results in these two studies. tResults and Discussion Hatchability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . _ .. Vitamin E and EmbryonicEye Development . . . . , . .. Vitamin E Analyses of Eggs . . . . . . . . . . . . . . . . . . . . .. PPROXIMATELY 3% T0 4 MILLION turkey breed- , er hens are maintained in the United States pr h year, with a total of 80 to 100 million poults ing hatched from eggs produced by these hens. Vt an average value of 75 cents per poult, the ex- _ ted income would be 60 to 75 million dollars _p nually from this phase of the 350 million dollar rkey industry. Approximately 65 to 75 million rkeys, consuming approximately 3 million tons feed, are raised annually from these poults. c The feeding of turkey breeder hens for max- um egg production, fertility and hatchability f» changed considerably during recent years as result of research in this field. The hatcha- lity of eggs from turkey hens usually is expect- _i to be fairly good during the first 8 to 12 eeks of the production period. Since hatcha- ility and fertility normally decline after this me, it has not been considered economically de- rable to maintain breeder flocks for a longer riod of time. " Nestler et al. (1936) reported that an all- getable protein diet would not maintain high tchability unless the diet was supplemented 'th pork liver meal, dessicated meat meal, fish eal or dried buttermilk. These workers also ported that dried whey would not maintain high tchability. Dickens et al. (1941) later reported at distillers dried grains with solubles would Yr replace dried skim milk, fish meal or meat ' raps in the New England Conference laying ra- on for turkeys as far as hatchabiltiy was con- rned. Stadelman et al. (1950) reported that tamin D had no effect on egg production but at the absence of vitamin D from the turkey eeder diet resulted in a decreased hatchability. Y was also shown by these Workers that the level i vitamin D supplied to dams had a direct in- uence on both body weight and calcification of ‘eebones of progeny that received no vitamin D ‘i four weeks of age. Jensen (1953) found that tamin E was necessary for high hatchability of rkey eggs. This was substantiated by Adkin- ii et al. (1955). A-,36-_percent increase in hatch-- ility was produced. by the addition of D-alpha- V! opheryl acetate to an all-vegetable protein diet espectively, associate professor, assistant professor, re- earch assistant, research assistant and head, Depart- a ent of Poultry Science; and professor, Department of oultry Science and Biochemistry and Nutrition. ihe Efiect ot Unidentified Factor Sources, Hntihiotics and D-a-tocooherul Hcetato on the Reproductive Performance oi Broad Breasted Bronze and Beltsville Small lJJhite lorlreos T. M. FERGUSON, R. L. ATKINSON, G. L. FELDMAN, C. H. WHITESIDE, J. H. QUISENBERRY and J. R. COUCH* fed Beltsville Small White turkey hens. Scott et al. (1955) found that supplements of 5 or 10 mg. of D-alpha-tocopheryl acetate per pound of feed would improve the hatchability in both a practical and semipurified diet. Each of these three reports indicated that natural feed ingred- ients may not contain sufficient vitamin E for the maintenance of high hatchability of turkey eggs. Jensen and McGinnis (1956) found that between 12 and 24 I.U. of vitamin E per pound of feed were required as a supplement to normal fee? ingredients in order to maintain adequate hatchability during an 18-week study. These investigations have been conducted with various types of turkeys and under varied ex- perimental conditions. Ingredients known as unidentified factor sources have been included in many combinations in an attempt to improve the hatchability of turkey eggs. These substances include such feedstuffs as condensed fish sol- ubles, alfalfa meal, dried brewers yeast, dried whey, grass juice and distillers dried solubles. This bulletin presents a review of the work done in turkey breeder hen nutrition at the Texas Agricultural Experiment Station since 1950. These investigations include tests with Broad Breasted Bronze and Beltsville Small White tur- keys reared to maturity on the range or on Wire floors. Various combinations of unknown hatch- ability factor sources, vitamins, antibiotics and antioxidants have been fed in an attempt to im- prove the reproductive performance of the tur- key breeder hen. PROCEDURE The turkeys used in the experiments being re- ported were from Broad Breasted Bronze or Belts- ville Small White stock of the Department of Poultry Science and the Texas Agricultural Ex- periment Station. All experiments were conduct- ed at the Texas A. and M. College Poultry Farm, College Station, Texas. All Broad Breasted Bronze experiments were conducted with groups of 12 to 17 birds per group, maintained in pens with wooden or concrete floors covered with 2 to 4 inches of cane pulp litter. The Beltsville Small White turkeys were reared to maturity in batteries and on wire floors to pre- vent access to droppings. Some experiments with Beltsville Small White hens were conducted on 3 wire floors, and in some tests birds were kept in individual cages throughout the laying period. Feed and water were supplied ad tibitnm. Arti- ficial insemination with pooled semen was used in the cage studies. Natural matings or a com- bination of natural and artificial matings were used in the other experiments. Eggs were gathered daily, stored in a cooler at 55° F. and set at weekly intervals. Eggs were candled at 7, 14, 21 and, 24 days of incubation. All eggs removed in candling were broken and the time of embryonic death was estimated. Embryos removed in candling and those failing to hatch were examined for gross changes in appearance. In some of the studies, live embryos were removed and preserved in an appropriate solution for mi- _ croscopic studies of the tissues. Eggs were ana- lyzed for tocopherol content at various stages of the experimental periods by Distillation Products Industries, Rochester, New York. Specific alterations in these above procedures are indicated in the discussion of individual ex- periments or treatments. Since D-alpha-toco- pheryl acetate is a compound used in feeds to pro- vide vitamin E activity, this compound often is used interchangeably with vitamin E in the dis- cussion which follows. Substitutions of the sup- plements to the all-vegetable-protein basal diet were made at the expense of corn or milo and soy- bean oil meal in order to maintain the same calcu- lated protein level within any given experiment. Hatchability data in all tables refer to percentage hatch of fertile eggs. RESULTS AND DISCUSSION Hatchability Experiment 1. Broad Breasted Bronze turkeys l‘ were reared to maturity in batteries on an all- vegetable protein diet (Atkinson and Couch, 1951 a,b). At 24 weeks, the hens were divided into groups of four birds each and placed in individual laying cages. One group received the basal (all- vegetable protein) diet unsupplemented. Supple- ments of 25 mg. aureomycin per kilogram, 66 mg. streptomycin per kilogram plus 13 mcg. vitamin B12 per kilogram, six percent fish meal, 2 per- cent aureomycin APF or 4 percent Liver “L” re- spectively, were added to the basal (all-vegetable protein) diet and fed to various groups of hens. One group of hens received weekly injections of vitamin B12 (10 mcg. per bird per week). The vitamin B12 was supplied in a crude form since it was not commercially available in pure quantities at the time of this experiment. The aureomycin APF was a combination of aureomycin and the Animal Protein Factor (APF) which was the term applied to vitamin B12 at this time. Egg produc- tion was increased considerably by supplementing the basal ration with aureomycin (25 mg. per kg.) or liver “L” (4 percent). Hatchability was very low (16 percent) in the basal group, and was im- proved considerably by various supplements as follows: aureomycin (25 mg. per kg.) 43.5 per- 4 cent, vitamin B12 injected (10 mcg. per b‘ l week) 53.8 percent, fish meal (6 percent) . and liver “L” (4 percent) 66.1 percent. Experiment 2. Atkinson et at. (1953) re a definite improvement in hatchability of) from Beltsville Small White turkey hens 1 had been reared to maturity on wire and ‘ all-vegetable protein diet, by ythe addition percent Biopar C to the basal ration. Bel ‘Small White turkeys were maintained on from hatching to maturity. At maturity \* were placed in a laying house with a wire; and were mated with Beltsville Small Whit which had been reared on range. Egg prod I was normal and fertility averaged approxi 90 per cent. Hatchability was poor (40 pe and steadily decreased to approximately 1 cent by the eighth week. _ After 10 weeks the house was divided in pens of 30 birds each and 5 percent Biop substituted for 5 percent milo in the ration ~. pen. Egg production remained normal i; tility was maintained at a high level (90 ; percent). Hatchability remained low (10 ; percent) for the basal group throughout w, tire experimental period. The addition of i_ C (5 percent) to the basal diet caused a s’ cant and immediate improvement in hatch, (50 to 60 percent) which was maintained f' proximately 8 weeks after which time a s decrease Was noted. - A number of embryos from the basal r: which died between the 24th and 28th day ‘ cubation or were alive but unhatched on the? day had shortened legs, hemorrhages und’ skin and a bulging eye condition in which th and/ or cornea appeared cloudy. These con 1 were thought to be due to a deficiency o known hatchability factors of which Biol‘ appeared to be a fairly good source. Experiment 3. Couch et at. (1954) cond a study to determine the value of distillers _ solubles and/ or fish meal as supplements to ' key breeder ration. Apparently there =_ beneficial effect on hatchability or fertility 2 the hens were fed either fish meal or dis dried solubles singly or in combination as s ments to the basal ration. Fertility rem at a fairly high level (65 to 90 percent),' groups throughout the 18-week experimen = riod. Hatchability declined to levels of 0 , percent after the 12th week of the experim, period. Egg production was slightly imp when both fish meal and distillers dried sol were included" in the diet but was not main 4, at a desirable level in any of the experim groups. This study shows clearly the nee more information regarding the nutrient req ments of the turkey breeder hen. ‘ Experiment 1,. The effect of adding dehyd alfalfa meal, condensed fish solubles, dried ~ and distillers dried solubles to the basal, TABLE 1. BASAL DIET I: Ingredient‘ Percent iund yellow corn 35 und milo - 34 bean oil meal 25 -- cium phosphate 3 ter shell flour 2.5 f .5 _plements added per lb. oi diet: 160 mg. MnSO. (Tech- grade). 4.500 LU. vitamin A. 1.200 I.C.U. vitamin D5. mg. riboflavin. 12.5 mg. calcium pantothenate. 20 mg. cin. 400 mg. choline chloride, 6 mcg. vitamin B12- able 1) on fertility, hatchability and tocopher- content of yolks of Broad Breasted Bronze tur- 1 eggs is shown in Table 2 (Atkinson et at. 255). The hatchability of fertile eggs from these ups of Broad Breasted Bronze turkey hens ged from 76.0 to 85.9 percent during the first weeks 0f the 17-week experiment and was not nged appreciably by the addition of dehydrat- alfalfa meal, condensed fish solubles, dried 5y. Qlistillers dried solubles or combinations of ise supplements to the basal diet (Table 1). ‘e basal diet included 2.5 mg. bacitracin, 1 mg. icillin and 2.5 mg. terramycin per pound of Fertility for the first 11 weeks apparently s not affected by dietary supplements (Table p. There was a rather low fertility (47.5 per- t) in the group fed alfalfa meal, dried whey 1 fish solubles (Table 5). There was no ap- ent explanation for this low fertility. During -. 12th through the 17th weeks of the experi- ntal period, a 23.3 percent increase in hatcha- ity of fertile eggs over that of the basal group _E 2. EFFECT OF ALFALFA, FISH SOLUBLES. DRIED 5| Y AND DISTILLERS DRIED SOLUBLES ON FERTILITY. TCHABILITY AND TOCOPHEROL CONTENT OF YOLKS I OF BROAD BREASTED BRONZE TURKEY EGGS Weeks Mp Sufiaplerngnttsl to 0'11 1 2'17 Egg‘: l as“ 1e — — — Percent — -— —°l/egg Fer- Hatch- Fer- Hatch- Y°1k2 tility ability tility ability mcg- §None 19.4 59.1 45.5 91.0 590 W 55% DAM 12.5 50.9 55.5 54.5 510 , 5% DAM + 5% CFS 14.5 15.0 90.9 51.0 455 5% DAM + 5% DW 52.5 52.1 59.5 55.4 515 _ 5% DAM + 3% ow + 5% crs 41.5 50.1 44.0 54.5 549 5% DAM + 5% DDS .151 55.9 55.5 54.0 1.019 5% DAM + 5% nos + CFS 51.5 55.0 55.5 15.5 151 5% DAM + 5% CFS + 9% ow + 5% nos 51.5 52.4 19.2 15.0 599 = dehydrated alfalfa meal; CFS = condensed fish 1 bles; DW = dried whey.- DDS = distillers dried solubles. opherol content oi egg yolks collected at the end oi the 1| week. was obtained when dehydrated alfalfa meal was added to the basal diet. A combination of dehy- drated alfalfa meal and condensed fish solubles added to the basal ration improved hatchability 50 percent above that obtained with the basal group during this period. A 9.7 percent increase was obtained with distillers dried solubles and dehydrated alfalfa meal added as supplements to the basal ration over that obtained with feeding alfalfa meal alone. In each instance the combi- nation of condensed fish solubles with one of the other unidentified factor sources produces an increase in the hatchability of fertile eggs (Table 2), above that obtained with the unidentified fac- tor source alone. The fertility of eggs from hens fed the basal diet was only 48.6 percent during the final 6- week period of the experiment (Table 2). In the remaining groups, except for group 5 as previ- ously mentioned, the fertility was greater in the groups fed condensed fish solubles and dehydrat- ed alfalfa than in the groups fed dried whey and distillers dried solubles supplemented with alfalfa meal. The various supplements which were added to the diets of the hens had very little influence on embryonic mortality during the first 11 weeks of the experimental period. However, embryonic mortality during the first week of incubation in the unsupplemented group increased from 6.3 percent during the first 11 weeks to 44.8 percent (Table 3) during the last 6 weeks. An increase in embryonic mortality (8.5 percent to 20.7 per- cent) was observed in the basal group during the last 4 days of the incubation period, between the 12th and 17th week of the experimental pe- riod. The addition of the unidentified factor sources to the diet of the hens in groups 2 through 8 decreased embryonic mortality during the ex- perimental period (Table 3). ' Embryonic mortality during the 24th to the 28th day of incubation for the last 6 weeks of the TABLE 3. EFFECT OF ALFALFA. FISH SOLUBLES. DRIED WHEY AND DISTILLERS DRIED SOLUBLES ON EMBRYONIC MORTALITY IN BROAD BREASTED BRONZE TURKEY EGGS DURING THE 12TH THROUGH THE 17TH WEEK OF EX- PERIMENT Times of embryonic deaths Supplement to Group b a s a1 dietl 0-7 days 24-28 days Mortality rate. percent 1 None 44.8 20.7 2 5% DAM 14.2 30.7 3 5% DAM + 5% CFS 13.8 5.9 4 5% DAM + 3% DW 26.9 12.4 5 5% DAM + 3% DW + 5% CFS 20.9 11.3 6 5% DAM + 5% DDS 19.8 14.0 7 5% DAM + 5% DDS + 5% CFS 12.3 9.8 8 5% DAM + 5% CFS + 3% DW + 5% DDS 9.5 15.5 ‘DAM = dehydrated alialia meal: CFS = condensed fish solubles." DW = dried whey: DDS = distillers dried solubles. 5 TABLE 4. EFFECT OF D-ALPHA-TOCOPHERYL ACETATE AND UNIDENTIFIED FACTOR SOURCES ON EGG PROD *1 FERTILITY, HATCHABILITY AND TOCOPHEROL CONTENT OF EGG YOLKS OF BELTSVILLE SMALL WHITE TURKEY) - » Production. Fertility. Hatch fertile Tocoph 1 Group Supplement to basal diet ,0 Eggs set 7° eggs‘ % You‘, 1 None 48.3 202 98.5 54.3 2 D-alpha-tocopheryl acetate (20 mg./lb.) 34.9 198 98.5 88.2 3 Fish solubles (5%) 38.6 187 65.8 47.2 4 Fish solubles (5%) + D-alpha- tocopheryl acetate (20 mg./lb.) 37.0 198 96.0 f. 93.2 5 Dried whey (8%) 53.1 15s 9a.? 51.5 6 Dried whey (3%) + D-alpha-tocopheryl acetate (20 mg./lb.) 39.7 167 92.8 85.8 7 Dried whey (3%) + fish solubles (5%) 44.4 174 99.4 52.0 8 Dried whey (3%) + fish solubles (5%) + D-alpha-tocopheryl acetate (20 mg./ lb.) 33.7 172 97.7 83.9 Effect of supplement other than D-alpha-tocopheryl acetate 1.2 None 41. 400 98.5 71.1 3.4 Fish solubles (5%) 37.8 385 81.3 75.1 5-6 Dried whey (3%) 35.4 32s 95.7 58.6 7.8 Fish solubles (5%) + dried whey (3%) 39.1 346 98.6 67.7 Effect of D-alpha-tocopheryl acetate 1.3.5.7 None 41.3 722 90.3 51.7 2.4.6.8 D-alpha-tocopheryl acetate (20 mg./lb.) 36.3 735 96.3 88.0 ‘Data from lst to 9th week. inclusive. Tocopherol content is of eggs collected during the ninth week. test varied from 5.9 to 30.7 percent (Table 3). The embryonic mortality in group 2, fed 5 per- cent dehydrated alfalfa meal only, Was greater than the basal group during this part of the in- cubation period. When alfalfa was combined with condensed fish solubles, the embryonic mor- tality was reduced to 5.9 percent. Jacobs et al. (1953) reported that dehydrated alfalfa meal increased the hatchability and fer- tility of eggs from chicken hens, when fed in the presence of vitamin B12. These data suggest that dehydrated alfalfa meal and condensed fish solu- bles contain greater amounts of the unidentified factors necessary to maintain normal turkey fer- tility and hatchability after 11 Weeks of produc- tion than distillers dried solubles or dried whey. Experiment 5. Supplements of D-alpha-toco- pheryl acetate, condensed fish solubles and dried Whey were added to the basal diet (Table Beltsville Small White turkey hens which: been reared to maturity on wire floors (' son et al. 1955). Vitamin A was increas 9,000 I. U. per pound at the start of the 17- experimental period and menadione was add a level of 2 mg. per pound. D-alpha-tocop acetate additions to the rations were rev after 9 weeks of the test period (Tables 4% 5). The addition of 20 mg. of D-alpha-tocop. acetate per pound of feed increased hatcha approximately 36 percent the first 9 Weeks (' 4). When the hatchability data (Table 4) 1 treated statistically by analysis of varia significant difference was found to exist be y, treatments. An F value of 13.82 Was 0b ” for treatments whereas only 3.02 was need TABLE 5. EFFECT OF D-ALPHA-TOCOPHERYL ACETATE AND UNIDENTIFIED FACTOR SOURCES ON EGG PRODU FERTILITY. HATCHABILITY AND TOCOPHEROL CONTENT OF EGG YOLKS OF BELTSVILLE SMALL WHITE TURKEY‘ Group Supplement to basal diet Pmdflfm“ Eggs set Ferizljty’ Haet;:si‘e%le Toccgglll; 1 D-alpha-tocopheryl acetate (20 mg./lb.) 39.5 164 89.6 63.3 2 None 31.1 108 99.1 79.4 3 Fish solubles (5%) + D-alpha- 28.2 112 96.4 e 72.2 tocopheryl acetate (20 mg./lb.) 4 Fish solubles (5%) 24.7 82 98.8 80.2 5 Dried whey (3%) + D-alpha-tocopheryl acetate (20 mg./lb.) 28.7 102 37.3 65.8 6 Dried whey (3%) 37.7 145 55.9 71.6 7 Dried whey (3%) + fish solubles (5%) + D-alpha-tocopheryl acetate (20 mg./ lb.) 27.8 89 74.2 75.8 8 Dried whey (3%) + fish solubles (5%) 28.7 123 44.7 85.5 ' Effect of supplement other than D-alpha-tocopheryl acetate 1,2 None 35.3 272 93.4 70.1 3.4 Fish solubles (5%) 26-5 194 97-4 75-7 5,6 Dried whey (3%) 33.3 247 48.2 69.7 7.8 Fish solubles (5%) + dried whey (3%) 28.3 212 57.1 80.2 Effect of D-alpha-tocopheryl acetate ‘ 1.3.5.7 D-alpha-tocopheryl acetate (20 mg./ lb.) 31.1 467 76.9 68.5 2,453 None 30.6 458 70.7 78.7 ‘Data from 10th to 17th week, inclusive. Tocopherol content is of eggs collected during the 17th week. 6 l. cance at the 0.01 level of probability. On analysis an F value of 90.35 was obtained l min E versus no vitamin E, Whereas, only as needed for significance at the 0.01 level bability. This indicated that under the 'ons of the experiment vitamin E was re- to maintain an optimum level of hatcha- _ hability of eggs from groups not fed sup- p. tal vitamin E during the first 9 weeks 3| 1, 3, 5 and 7) was increased approxi- 17 percent by supplementation of the - ith D-alpha-tocopheryl acetate at the be- g of the 10th week of the experimental l ,(Table 5). j average hatchability of eggs from groups 6 and 8 was 88 percent (Table 4) for the 59-week period, during which time all birds i" groups received a supplement of 20 mg. lpha-tocopheryl acetate per pound. Remov- g the D-alpha-tocopheryl acetate from the diet e end of the 9th week caused the hatchabili- decrease from 88 to 78.7 percent (Tables 4 .5). i? embryos found dead during the incubation w were examined in order to determine time leath and to check for any abnormalities 9 might have occurred. There were 309 embryos out of 652 fertile eggs during the _ 9-week period from hens that did not re- p D-alpha-tocopheryl acetate; however, there _ only 83 dead embryos out of 708 fertile eggs " hens fed D-alpha-tocopheryl acetate. A- l number of embryos from the unsuppliment- oups died between the 24th and 28th days cubation. This was not true for those groups lemented with D-alpha-tocopheryl acetate. eficient embryos dying after the 17th day of ‘Nation were found to be smaller than normal [etermined by body measurement. Most of deficient embryos which died during incuba- j appeared to be blind and were found to have ‘udy lens or a cloudy spot under the cornea. ‘i of the embryos were found to have both of f conditions. ilure to obtain a rapid reversal of hatchabil- iupon reversal of the vitamin E treatments ble 5) indicates that the turkey hen may be '__ to store vitamin E in the body tissues to a ‘ter extent than was indicated by the report ensen (1953). He reported that the hatcha- of eggs from birds fed D-alpha-tocopheryl te was reduced from 80 to 45 percent by the oval of the supplemental vitamin E from the for 3 weeks. _ ithin 2 weeks affter the diets were reversed, above-mentioned’ abnormalities ceased to oc- l and none were found in any group during the i 6 weeks of the 17-week experimental period. 's would also indicate thatthere was consider- storage of the vitamin in the tissues of the @- It is apparent from Table 4 that the addition of D-alpha-tocopheryl acetate to the diet of tur- key hens had increased the tocopherol content of egg yolks by the end of the first 9 weeks of the experiment. Reversal of the diets at the begin- ning of the 10th week resulted in a reversal, also, of the tocopherol content of the egg yolks, at the end of the 17th week (Table 5) of the experiment- a1 period. Such a decrease in the tocopherol con- tent of the egg yolk did not result in as low a percentage hatchability in eggs from groups 2, 4, 6 and 8 as had been observed earlier in eggs from groups 1, 3, 5 and 7, even during the final week of the experiment. No explanation is immediate- ly apparent for the hatchability remaining at a higher level in groups 2, 4, 6 and 8 since the toco- pherol content of the eggs had decreased to a point where much lower hatchability might have been expected. The average hatchability of eggs from groups 2, 4, 6 and 8 was 78.7 percent from the 10th to the 17th week of the test, while it was 51.7 percent for eggs from the same birds during the 9 weeks prior to reversal of the diets. It is possible that unidentified factors, independent of or interacting with alpha-tocopheryl acetate, may have caused the hens to be depleted more slowly of tocopherol stores. Late in the season 200 Broad Breasted Bronze turkey eggs were obtained from a commercial turkey breeder who was feeding a commercial turkey breeder mash and had experienced a con- siderable drop in both fertility and hatchability. These eggs were incubated in our laboratory. Only 30 percent of the 200 eggs were found to be fertile, and only 41.7 percent of the fertile eggs hatched. Eight of the embryos which died between the 24th and 28th day of incubation were found to have a cloudy lens, were smaller in size and were quite similar in appearance to embryos obtained from hens in experiment 5 that had not received supplemental D-alpha-tocopheryl acetate. Experiment 6. Broad Breasted Bronze turkeys were fed the basal diet (Table 1) unsupplemented and supplemented with Vitamin E (D-alpha-to- copheryl acetate), dehydrated alfalfa meal, con- densed fish solubles and combinations of these substances, (Table 6) (Ferguson et al. 1956). The basal ration used was the same as shown in Table 1 except that an additional 4,500 I. U. of vitamin A were added per pound of ration. These hens were reared on range and had been fed a practical type ration containing sources of unidentified factors. The hens were maintained on litter during the experimental period, rather than on wire floors, in order to determine the effectiveness of feeding supplements of unidenti- fied factor sources and vitamin E to turkey hens maintained under practical conditions. The addition of Vitamin E, dehydrated alfalfa meal and condensed fish solubles to the basal diet resulted in hatchability varying from 71 to 79 percent during the first 8 weeks of the test (Table 7 TABLE 6. HATCHABILITY OF FERTILE EGGS AND TOCO- PHEROL CONTENT OF EGGS FROM BROAD BREASTED BRONZE TURKEY HENS Hatchabilit Toco herol _ Supplements (%) Y (nwq-gyolk)’ BT12? Group to basal diet‘ 1'3 9-20 13 21 H393} wks. wks. wks. wks. tahlY 1 None 74 56 430 540 24.3 2 Vitamin E 77 60 1.820 920 30.4 3 DAM 71 60 520 370 26.3 4 DAM + ' vitamin E 79 68 630 1.250 21.6 5 CFS 76 68 520 840 19.9 6 CFS + vitamin E 78 68 1.170 1.100 18.8 7 CFS + DAM 74 68 690 580 18.5 8 CFS + DAM + vitamin E 76 77 1.180 1.050 8.0 ‘Vitamin E = D-alpha-tocopheryl acetate. 20 mg./1b.: DAM = dehydrated alfalfa meal 5%." CFS = condensed fish solubles 5%. 3Pre-experimental tocopherol levels: 471 mcg./yolk. “Fourth week embryonic mortality in eggs laid during weeks 9th to 20th. 6). In each instance where vitamin E was added (groups 2, 4, 6 and 8) hatchability Was increased 2 to 8 percent over that observed in groups 1, 3, 5 and 7. The latter groups represent the groups fed unidentified factor sources without supple- mental vitamin E. The hatchability of eggs during the last 12 weeks of the experiment was improved from 4 to 12 percent by the addition of either vitamin E, dehydrated alfalfa leaf meal or condensed fish solubles, fed singly or in combination of any two of these supplements. However, when all three TABLE 7. EFFECT OF DEHYDRATED ALFALFA MEAL. CONDENSED FISH SOLUBLES AND DRIED BREWERS YEAST ON THE HATCHABILITY OF BROAD BREASTED BRONZE TURKEY EGGS 0-11 weeks 12-17 weeks 12-24 weeks Supplement Group to Total % Total % Total % basal diet eggs hatch- eggs hatch- eggs hatch- set flbllllYl set ability‘ set ability‘ None 428 75.0 182 24.8 352 36.9 5% dehydrated alfalfa meal 525 82.9 161 62.23 311 55.9 10% dehydrated alfalfa meal 562 77.5 167 63.03 284 67.03 5% condensed fish solubles 478 75.9 120 _ 73.53 246 56.8 5% condensed fish solubles + _ 5% dehydrated alfalfa meal 673 83.8 208 78.03 378 73.33 6 5% condensed fish solubles + 10% dehydrated alfalfa meal 558 82.9 194 71.23 300 62.43 7 5% dried brew- ers yeast 593 74.4 200 64.13 383 57.9 8 5% dried brew- ers yeast + 5% condensed fish solubles 593 84.6 254 78.63 462 74.53 UIbODNHH ‘Percent of fertile eggs. “Significant at .05 level. “Significant at .01 level. 8 of the supplements were fed (group 8),: cent hatchability was maintained Whi L sented an increase of 21 percent over t w’ unsupplemented group (group 1). The» this group (group 8) were the only on did not show a decrease in hatchability = the 9th and the 20th week of the expe period. Analysis of the dataEfor the 9 t0 I period showed that the addition of vi -§ (D-alpha-tocopheryl acetate) to the dieti an increased hatchability which was sta significant at the 5 percent level of pro y, The feeding of condensed fish solubles p, an increase in hatchability which was si_ , at the 1 percent level of probability. e creased hatchability due to vitamin E is i ment with reports by Jensen (1953), ' g et al. (1955) and Ferguson et al. (1954).; Embryonic mortality was essentially pf for all groups during the first 8 weeks. was also very little difference in the em, mortality between groups during the _, weeks of the incubation period for eggs :1 ing the 9th through the 20th week. _ percent embryonic mortality was found”. the 4th week of incubation of eggs from ‘ in group 8 which had been fed supple p alfalfa meal, fish solubles and vitamin E’ pha-tocopheryl actate) (Table 6), while v bryonic mortality ranged from 18.5 to Iii cent for the remaining groups duringt ' period. Experiment 7. Feldman et al. (1957a) results of supplementing an all-vegetable ration with dehydrated alfalfa meal, dri_ ers yeast and condensed fish solubles. reared Broad Breasted Bronze turkeys w, in this study, and the hens were mainta! litter during the laying season. Vitamin; mg. per pound), chlortetracycline (20 . pound), penicillin (5 mg. per pound) .5 thyl naphtho quinone (2 mg. per pound) W added to the basal diet shown in Table 1. dition, the levels of dicalcium phosphate ter shell flour were changed so that each ed at a level of 3.5 percent. The addition cium phosphate and oyster shell flour Wer at the expense of corn. r. The hatchability of fertile eggs from f Breasted Bronze turkey hens varied fro percent to 84.6 percent during the first 1; of the 24-week study. The hatchability of eggs from hens fed diets supplemented w‘ densed fish solubles in combination with‘ dehydrated alfalfa leaf meal or dried t yeast was observed to be slightly higher ti of hens fed the unsupplemented basal rati hens fed rations supplemented with onl. drated alfalfa meal or dried brewers yeast 7 . 1 The hatchability of fertile eggs from H‘ the unsupplemented basal diet (group 1) ={ a decided drop which was evident by the end " he 12th week. The hatchability for this group raged only 24.8 percent between the 12th and h week of the study. The addition of dehy- ted alfalfa meal, condensed fish solubles or brewers yeast to the" basal ration or con- I sed fish solubles fed in combination with de- rated alfalfa meal or dried brewers yeast pro- Yed a significant increase in the hatchability fertile eggs during this same period. Dehy- ted alfalfa meal was the only supplement 'ch was fed at more than one level. A- l0-per- t level of this supplement failed to produce any her increase in the hatchability of fertile eggs the 5-percent level from the 12th through 17th week. There was an indication from the data in Table that a depletion of the hatchability factor(s) ‘3urred between the 12th and 17th weeks in "ups fed supplements of 5 or l0 percent dehy- ted alfalfa meal or 5 percent dried brewers »:st. The highest hatchability of fertile eggs was ob- _;u by supplementation of the basal diet with binations of 5 percent condensed fish solubles i» 5 percent dehydrated alfalfa meal (group 5) with 5 percent condensed fish solubles and 5 cent dried brewers yeast (group 8). These ferences were not found to be statistically sig- icant. The hatchability of fertile eggs from Broad gsted Bronze turkey hens fed diets contain- _}'£ 5 percent dehydrated alfalfa meal (group 2), ercent condensed fish solubles (group 4) or 5 cent dried brewers yeast (group 7) decreased l further as shown in Table 7 for the period .1 'nning with the 12th week and ending with (24th week. Dehydrated alfalfa meal (group ' at the 10-percent level, on the other hand, was le to maintain hatchability at a satisfactory el during this same period. This would indi- A - that 5 percent dehydrated alfalfa meal did f- supply sufficient quantities of the unidenti- in hatchability factor(s). Hens fed 5 percent ydrated alfalfa meal, 5 percent condensed solubles and 5 percent dried brewers yeast 'bited almost identical average hatchability for g period of the 12th to the 24th week. ‘ Highly significant increases in the hatchability fertile eggs were obtained in groups 5 and 8. ese hens were fed combinations of condensed h solubles (5 percent) and dehydrated alfalfa . (5 percent) and condensed fish solubles (5 cent) and dried brewers yeast (5 percent) as wn in Table 7. * jTWo principal peaks of embryonic mortality re observed in eggs from the hens in this study. A. first occurred during the‘ first 7 days of the = ubation period and the second occurred during a last 7 days of the incubation period. TABLE 8. CONDENSED FISH SOLUBLES AND DRIED BREWERS YEAST ON THE EMBRYONIC MORTALITY OF BROAD BREASTED BRONZE TURKEY EGGS EFFECT OF DEHYDRATED ALFALI-‘A MEAL. First week Fourth week Supplement incubation incubation Gm“? ‘° 0-11 12-17 12-24 0-11 12-17 12-24 basal diet wks. wks. wks. wks. wks. wks. — —— Embryonic mortality. percent — — 1 None 8.9 70.0 48.2 13.4 2.9 13.1 2 5% dehydrated alfalfa meal 7.0 24.8 28.9 9.2 11.4 13.3 3 10% dehydrated alfalfa meal 7.0 13.9 14.9 14.1 25.3 18.4 4 5% condensed fish solubles 10.3 19.5 35.5 8.9 6.2 6.0 5 5% condensed " fish solubles + 5% dehydrated alfalfa meal 3.3 13.7 17.9 5.6 6.3 7.5 6 5% condensed fish solubles + 10% dehydrated alfalfa meal 9.5 14.1 26.5 7.3 10.8 9.0 7 5% dried brew- ers yeast 5.7 21.7 28.0 15.7 13.0 12.5 8 5% dried brew- . ers yeast -|- 5% condensed fish solubles 5.8 13.6 15.3 7.6 7.0 8.2 Slight differences in embryonic mortality were observed (Table 8) during the first 11 weeks of the experimental period which could not be at- tributed to the dietary treatments. Supplementa- tion of the basal diet with 1O percent dehydrated alfalfa meal reduced the first week embryonic mortality below that obtained on the diet supple- mented with 5 percent dehydrated alfalfa meal from the 12th through the 17th week. Condensed fish solubles fed in combination with either 5 per- cent or 10 percent dehydrated alfalfa meal re- duced the first week embryonic mortality below that of the condensed fish solubles group. The feeding of 5 percent dried brewers yeast was no more effective in reducing the mortality of em- bryos during this period than was 5 percent dehy- drated alfalfa meal or 5 percent condensed fish solubles, but when fed in combination with 5 per- cent condensed fish solubles, a slight improvement was noted. TABLE 9. TOCOPHEROL CONTENT OF EGG YOLKS FROM TURKEY HENS FED A GRAIN-TYPE OF DIET SUPPLEMENTED WITH AN ANTIOXIDANT. DIPHENYL-P-PHENYLENEDIAME (DPPD) VITAMIN E AND CONDENSED FISH SOLUBLES Diet‘ Weeks on diet Tocopherol per X yolk. mcg. Pre-experimental 5 1360 Basal 10 430 Basal -|- E 10 925 Basal -|- DPPD 10 470 Basal + E + DPPD 10 990 Basal + E’ 4 1135 Basal -|- E -|- CFS’ 4 1360 ‘D-alpha-tocopheryl acetate (vitamin E): added at level of 20 mg./lb.; DPPD added at 0.25 lb. per ton. QSee text for complete explanation. The first week embryonic mortality, during the 12 to 24 Weeks of the test, was greatly reduced when the hens were fed diets containing 10 per- cent dehydrated alfalfa meal (group 3) , 5 percent condensed fish solubles and 5 percent dehydrated alfalfa meal (group5), or 5 percent condensed fish solubles and 5 percent dried brewers yeast (group 8). There were practically no differences in the 4th week embryonic mortality between experimental groups. However, it should be pointed out that in this case, the groups which were fed rations containing condensed fish solubles (groups 4, 5, 6 and 8) consistently exhibited the lowest 4th week embryonic mortality throughout the entire 24 weeks of the study. Experiment 8. Thirty-six Beltsville Small White hens from groups 2, 4, 6 and 8 of experiment 5 were maintained on the basal diet (Table 1), (Ferguson et al. 1954b). Experiment 5 had term- inated in July when the hens stopped laying. Af- ter about 8 weeks the hens were placed under lights and mated with toms similarly treated. The first eggs were laid approximately 12 weeks af- ter the hens had gone out of production. Analy- sis for tocopherol was carried out on eggs col- lected during the fifth week of a pre-experi- mental period. Hatchability of fertile eggs during this 5-week period was O, 45, 70, 56 and 66 per- cent, respectively. At this time the birds were divided into four groups and fed the basal diet supplemented with 2O mg. per pound vitamin E (D-alpha-tocopheryl acetate) and an antioxidant, 0.0125 percent DPPD (N, N’-Diphenyl-p-pheny- lenediamine) as shown in Table 9. Hatchability of fertile eggs from the basal group was only 25 percent for the following 10- week period. Supplementation of the basal diet Figure 1. Normal-sized crop (top leit) and gizzard (top right) from Beltsville Small White turkey, removed immedi- ately after hatching. Enlarged thin-walled crop (bottom left) and gizzard (bottom right) from 27-day embryo. The lower pictures were made oi preserved material and the gizzard is smaller than it appeared in the live embryo. l0 TABLE 10. EFFECT OF VITAMIN E AND DPPD S _d MENTS ON HATCHABILITY AND TOCOPHEROL CO g OF EGGS FROM BELTSVILLE SMALL WHITE TUB I1: Tocopher mcgJyo ‘l, 1-5 wks. 6-18 wks. 8 wks. 16 Supplement to ‘X, hatchability Group basal diet 1 None 38 . 43,48 335 2 Vitamin E i (20 mg./lb.) 81 77 910 3 DPPD (1/4 lb./ton) 58 68 130 4 Vitamin E (Z0 mg./lb.) —|- DPPD (1/4 11)./ton) 43 77 840 ‘Pre-experimental values, 260 mcg./ yolk. with Vitamin E (D-alpha-tocopheryl acetate)! creased hatchability to 50.8 percent. The -- tion of DPPD and a combination of DPPD. vitamin E resulted in hatchability values 0 percent and 36 percent, respectively. i During the 10-week experimental period, 3 incidence of embryonic mortality was high i; groups during the 4th week of incubation. l? embryos from all groups were characterize edema, frequent eye disorders, an enlarged thin-walled crop and gizzard (Figure 1), u deposits in the kidneys and ureters and L ionally, an enlarged heart. ; After 10 weeks 26 of the birds were div' y into two groups (Table 9). One group was the basal diet plus vitamin E, and the other g é received the basal diet plus condensed fish g ubles and vitamin E. The hatchability of th supplemented group and the fish solubles 11 vitamin E group average 77 and 78 percent, spectively. The incidence of embryonic ab, malities was not observed during the 4-week iod except for four instances; one in group 1f of 117 fertile eggs, and three in group 2 ou‘ 127 fertile eggs. Analyses of eggs for tocophg content during this experiment are shown: Table 9. Experiment 9. Beltsville Small White tur, hens, reared to maturity on wire floors, were the basal diet (Table 1) in which de-germed l (63-percent) replaced ground yellow corn ground milo (Ferguson et al. 1955b). Soy‘ oil meal was increased to 31 percent. One g i received the basal diet unsupplemented and '- other groups received the basal diet supplem ed as indicated in Tables 10 and 11. Hatchability was low during the first 5 i in all groups except the one supplemented vitamin E (D-alpha-tocopheryl acetate) (T . 10). Some improvement was noted in ha bility when DPPD was added, but a lowe hatchability occurred when DPPD was fed vitamin E. During the 6th through the 1 g week, the hatchability improved in all gro with those containing vitamin E giving the r hatchability. Tocopherol values of egg yolks w, not increased by the addition of DPPD or de drated alfalfa meal (Tables 10 and 11). 11. EFFECT OF VITAMIN E AND DEHYDRATED ALFA MEAL ON HATCHABILITY AND TOCOPHEROL NT OF EGGS FROM BELTSVILLE SMALL WHITE TURKEYS Tocopherol‘ Supplement to hatchability basal diet 12 wks. mggdgslflk None 40 350 Vitamin E (20 mg./lb.) 73 715 Dehydrated alfalfa meal (5%) 66 a 260 Vitamin E (20 mg./ lb.) + de- hydrated alfalfa meal (5%) 63 805 p. experimental values. 260 mcg./yo1k. _ atchability of eggs after 12 weeks (Table 11) g highest when vitamin E was added. An im- ‘vement in hatchability over the basal group A also obtained when dehydrated alfalfa meal included in the diet. mbryonic abnormalities similar t0 those re- ed in experiment 8 occurred in all groups. , xperiment 10. Whiteside et col. (1957) fed ad Breasted Bronze turkey hens the basal diet table 1) unsupplemented or supplemented with ercent dried brewers yeast (Table 12). Addi- nal groups received, in addition to the dried wers yeast (5 percent), either 5 percent de- l» rated alfalfa meal or 5 percent condensed fish ubles. One group received both dehydrated alfa meal (5 percent) and condensed fish sol- (es (5 percent) in addition to the dried brew- p yeast (5 percent). Supplements were added the expense of grain and soybean oil meal so j-to maintain the same protein level among the 'ious groups. The basal diet also contained icillin (5 mg. per pound), aureomycin (20 mg. i» pound), and furazolidone (4.5 mg. per "und). Vitamin E was not included as a supple- nt to the basal diet. All unidentified factor I rces increased hatchability above that obtain- when the basal diet was fed. The addition of dried brewers yeast to an all- etable protein basal diet, without supplement- j vitamin E (D-alpha-tocopheryl acetate), im- tved the hatchability of eggs by 19 percent. No iprovement was obtained by adding dehydrated alfa meal to the basal ration supplemented th dried brewers yeast. However, when dried "11.? 12. EFFECT or DRIED BREWERS YEAST. ALFALFA a connsnssn FISH SOLUBLES on TURKEY HATCH- ABILITY FROM THE s TO IS-WEEK PERIOD brewers yeast was fed in combination with con- densed fish solubles or with dehydrated alfalfa meal and condensed fish solubles, further in- creases of 6 and 8 percent, respectively, were ob- tained over the group fed dried brewers yeast. Hatchability records were kept only for 15 weeks. Egg production data for the 26-week experi- mental period, and the amount of feed required per egg are shown in Table 13. The number of eggs produced by all groups fed dried brewers yeast was greater than the number produced by the basal group (Table 13). From 11 to 16 more eggs per hen were produced when dried brewers yeast and other unidentified factor sources were included in the diet. The average feed intake per hen, to produce an egg, ranged from 1.12 to 1.29 pounds. A slight reduction is noted when dried brewers yeast is added. The small increases (above group 2) no- ted in groups 3 and 5 may be due to the lowered energy content of the feed as a result of includ- ing alfalfa in the diet. Vitamin E and Embryonic Eye Development A gross and microscopic study was made of the blind turkey embryos observed in experiment 5 (Ferguson et at. 1954). A cloudiness in the cen- tral portion of the lens was found among the em- bryos from hens fed the all-vegetable protein basal ration without added vitamin E. This con- dition was observed in both eyes of 13 out of 30 live embryos examined on the 29th day of incu- bation. These embryos pipped the shell but failed to hatch. Seven of these had hemorrhages in the vitreous humor. One or both of the eyes fre- quently had a bulging of the cornea, giving the embryo a protruding eye appearance in extreme cases (Figure 2). The cornea was found to be slightly irregular in some embryos when exam- ined with a binocular microscope. A yellowish- white spot of material was seen in the eyes, be- tween the lens and the cornea, in a few specimens. This condition also occurred at later stages of embryonic development. An irregular-shaped iris was often associated with the bulging cornea, which made the pupil likewise irregular. Upon TABLE 13. EFFECT OF DRIED BREWERS YEAST. DEHYDRAT- ED ALFALFA MEAL AND CONDENSED FISH SOLUBLES‘ ON THE EGG PRODUCTION OF BROAD BREASTED BRONZE TURKEY HENS FOR A PERIOD OF 26 WEEKS 1% hatch‘ HatdmbilitY Supplement Total Increase E I ~ Lb. ‘ su lemem i, : i‘ 11511111!‘ % lmirease Group to eggs - over ggs feed/ oup pp : 3-15 ‘We!’ bflsul- basal diet 26 wks. basal n egg i i‘ wks. 3-15 wks. 1 Basal 918 61 1.21 Basal 49 2 + DBY 1082 164 72 1.11 + DBY 68 +19 3 + DBY + alfalfa 1154 236 77 1.16 + DBY + Alfalfa 69 +20 4 + DBY + CFS 1124 206 75 1.13 + DBY + CFS 74 +25 5 + DBY + alfalfa + DBY + Alfalfa + CFS 76 +27 + CFS ‘ 1075 157 72 1.29 11 Figure 2. Turkey embryos, 27 days. Embryo with bulging eyes (left) from hen on E-deficient diet. Normal embryo (right) from hen fed a diet supplemented with vitamin E. dissection 0f the lens, pressure applied firmly to the outside resulted in its rupture with a split- ting out of a smaller miniature lens and the loss of a Watery fluid. In some instances, a still smallerlens-like structure could be forced out. The eyes of the embryos obtained from hens sup- plernented with vitamin E apparently were nor- ma . The results of a study of microscopic sections of the lens from each of the two groups of em- bryos are shown by camera lucida drawings in Figure 3. Figure 3A» is a section near the center of the lens of a 29-day embryo from a hen that was fed added vitamin E. A portion of the iris and ciliary body are included. Figure 3B is a similar section from near the center of the lens of an embryo of the same age where the hen did not receive supplemental vitamin E. The lens is Figure 3. Drawings of microscopic sections through the eye of the turkey embryo. Normal lens is shown in A. Degenerative changes in the lens are shown in B. 12 apparently the same diameter as 3A, but ter and has a small lens body inside the ou layer. Degenerative changes have o; throughout the lens, especially in the cent posterior portions. a Measurements of the principal skeletal p) live embryos from the supplemented and plemented groups were made. These e were 29 days of age, and had pipped the sf, did not hatch. The legs, wings and body of the embryos from the groups not suppl ditional D-alpha-tocopheryl acetate were the supplemented groups. Ten of the 301i; bryos examined from the unsupplemented i on the 29th day of the incubation period a an edematous area on the neck at the backqj head. This condition did not occur in the D‘ tocopheryl supplemented groups. ~ Experiment 11. A detailed pathological of the cataract found in turkey embryos of from hens fed a synthetic type diet deficit vitamin E was reported by Ferguson (1956). a Beltsville Small White turkey hens wet vided into two groups, placed in individual and fed the synthetic basal diet (Table 14).f{ man E (D-alpha-tocopheryl acetate), 20 r pound, was provided as a supplement group. After nine and one-half Weeks on thy), the vitamin E (D-alpha-tocopheryl acetate) was increased to 40 mg. per pound. i Turkey embryos from the basal group i; moved daily from the 5th through the 28 of incubation. Only living embryos were and preserved for histological study. t, were noted in the external appearance, le epithelium and cornea. These embryos showed a keratocon‘, change (a bulging of the eyes) at about 19 of incubation, which increased as the e aged. Hemorrhage within the eye was ob, in two of the deficient embryos. Opacities ' in size in the deficient embryos; however, t TABLE 14. SYNTHETIC BASAL DIET Ingredient‘ Sucrose Drackett C-1 assay protein Fresh lard Salts mixture2 lThe following ingredients were added per kg. of diet; thiamin. 6 mg. riboflavin. 15 mg. calcium pantothe, mg. niacin, 4 mg. pyridoxine. 20 mg. para-aminobenz, 3.800 I.C.U. vitamin D3. 10.000 I.U. vitamin A, 33 mg. 1.000 mg. inositol. 0.2 mg. biotin. 2 mg. folic acid. i menadione. 2.000 mg. choline chloride. 7.5 gm. me { 4 gm. glycine. 30 mcg. vitamin B12. 2The salts mixture supplied the following per kg. -- mg. manganese. 30 mg. iodine, 559 mg. magnesium. iron. 4.3 mg. copper. 6.7 mg. zinc. 0.11 mg. cobalt. f‘ mixture supplied 2.5% calcium. 1.0% phosphorus. 0.5 and 0.2% potassium. "Y l5. INCIDENCE OF EYE DISORDERS IN BELTSVILLE l WHITE TURKEY EMBRYOS FED A PURIFIED SYN- ; THETIC TYPE DIET Diet Opfecrsy in Keratoconus —E Z1 (38.8%) 9 (17%) +5 2 (3.6%) 0 p: were usually similar in the same ‘bird. There _e opacities in one or both eyes of 21 out of 54 _, min E deficient embryos (Table 15). Only 2 ryos with opacities were observed in 55 tur- “embryos from the vitamin E supplemented "P- he most conspicuous change present in the t of these embryos occurred in the lens, char- rized by a liquefaction of either a part or es- tially all of the lens protein. The earliest ilnges noted were small focal areas of liquefac- t located either in the center or at the peri- ry of the lens immediately beneath the cap- p Preliminary studies also showed degener- changes in tissues other than the eye, which i now under investigation. 5 f» rtificial insemination was used to obtain fer- ' eggs. Hens were inseminated weekly with ml. of pooled semen which had been diluted _ or 1:2 with 0.85 percent saline solution. The (rage fertility of eggs as a result of this method »= 90 percent over a 15-week period. The fer- A y of eggs from hens provided vitamin E was percent, and 85 percent fertility was obtained the group not provided vitamin E (Table 16). atchability of eggs from both groups (Table Was determined for 15 Weeks. In the group iving the basal synthetic diet, without vita- E, a 21 percent hatchability of fertile eggs _; obtained. In the supplemented group, hatch- _lity of eggs was 55 percent. It should be point- iout that this synthetic diet did not contain un- intified hatchability factor sources. Tocopherol ues of eggs from these hens are given in Table i Vitamin E Analyses of Eggs he importance of vitamin E as a factor in in- asing hatchability has been demonstrated in Any of these experiments. Analyses of the eggs F étheir alpha-tocopherol content revealed some resting facts. l- l6. HATCHABILITY AND FERTILITY OF EGGS FROM W HENS FED A SYNTHETIC-TYPE DIET (15 WEEKS) Diet i‘. No. eggs Ferigfity’ afifigllllk f =1 21v as 21 p1 + Vitamin E‘ 190 - 95 5s lpha-tocopheryl acetate added at level oi 20 mg./lb. tor eeks and 40 mg./lb. thereafter. Table 4 shows that in hens not provided supple- mental vitamin E, the average tocopherol content after 9 weeks was 200 mcg. per yolk, while 838 mcg. per yolk Was found in eggs from hens whose diets were supplemented with vitamin E. It was also found by these analyses that the alpha-to- copherol occurs only in the yolk and not in the egg white. A reversal of the vitamin E treat- ments after 9 Weeks (Table 5) showed that when vitamin E was removed from the diet, the toco- pherol content of the egg decreased. Tables 4 and 5 also show there was no effect on the toco- pherol content of the egg due to dietary supple- ments of fish solubles or dried whey. _ In experiment 8, the hens from groups 2, 4, 6 and 8 of experiment 5 were continued on the basal diet (Table 1) for 8 weeks. When these hens were brought back into production, it was found that the tocopherol content of the first eggs averaged 1,360 mcg. per yolk (Table 9), whereas the previous average was only 215 mcg. per yolk. As egg production progressed, the values de- creased to 430 and 470 mcg. per yolk in groups not provided supplemental D-alpha-tocopheryl acetate and to 925 and 990 mcg. per yolk in the E-supplemented groups, which was consistent with earlier results. These data indicate that the hen is capable of storing this vitamin during non- laying periods and that tocopherol storage is de- pleted as egg production continues. During the last few Weeks of the test, when all hens were get- ting vitamin E, egg production declined and the tocopherol content of the eggs again increased. Analyses of eggs from hens fed various combi- nations of dehydrated alfalfa meal, condensed fish solubles and dried brewers yeast (Table 18) at the end of the 17-week study showed that supple- mental vitamin E maintained the tocopherol con- tent of the egg at a high level throughout the lay- ing period. The pre-experimental level of toco- pherol was 700 mcg. per yolk for the range-reared birds. In all instances, the tocopherol was in- creased during the laying period. The value for group 5 is within the range of normal values ob- tained when supplemental D-alpha-tocopheryl ace- tate is added to the diet at a level of 20 mg. per pound. Values for the other groups showed that there was no consistent effect on the tocopherol content of the egg due to supplements of dehy- drated alfalfa meal, condensed fish solubles or dried brewers yeast. TABLE 17. TOCOPHEROL CONTENT OF EGG YOLKS FROM HENS FED A PURIFIED SYNTHETIC-TYPE BASAL DIET Tocopherol per yolk (mcg.)‘ D. let Pre-Exp. 5 wks. 13 wks. Basal 260 130 245 Basal + E‘ 260 797 130D ‘Vitamin E added at 20 mg./lb. for 9 weeks and 4O mg./lb. thereafter. ' l3 TABLE 18. TOCOPHEROL CONTENT OF EGG YOLKS Tocopherol/ “:2.” s"::::r::*;.*° mg- ' 17 wks.‘ 1 None 1595 2 5% dehydrated alfalfa meal 1150 3 10% dehydrated alfalfa meal 1755 4 5% condensed fish solubles 1090 5 5% condensed fish solubles + 5% dehydrated alfalfa meal 1025 6 5% condensed fish solubles +- 10% dehydrated alfalfa meal 760 7 Dried brewers yeast 1075 8 5% dried brewers yeast + 5% condensed fish solubles 1265 ‘Pre-experimental values. 700 mcg./yolk. In experiment 9 (Tables 10 and 11), the toco- pherol content of the eggs was increased only when D-alpha-tocopheryl acetate was present in the diet, and no increasewas noted with the addi- tion of dehydrated alfalfa meal or DPPD. The tocopherol content of the egg yolks was in- creased in experiment 6 when vitamin E (D-al- pha-tocopheryl acetate) was added to the diet (Table 6). The addition of condensed fish sol- ubles or dehydrated alfalfa leaf meal produced, essentially, no effect on the tocopherol content of the yolks. No explanation is readily apparent for the low value obtained in group 4 (13 weeks) or the slightly increased tocopherol level in group 5 (21 weeks) . In experiment 11 the hens had been reared to maturity on wire floors and had been fed an all- vegetable protein diet without supplemental vita- min E. The experimental diet was a synthetic one which contained no added vitamin E except for one group of birds. The addition of D-alpha- tocopheryl acetate to the diet increased the toco- pherol content of the egg, with higher levels of the vitamin resulting in a further increase in the FIGURE 19. EFFECT OF DEHYDRATED ALFALFA MEAL. CONDENSED FISH SOLUBLES AND DRIED BREWERS YEAST ON EGG PRODUCTION, AVERAGE DAILY FEED CONSUMP- TION AND AMOUNT OF FEED CONSUMED PER EGG PRODUCED OF BROAD BREASTED BRONZE TURKEY HENS Eggs/ Feed/ Feed/ Group Supplementation hen/ bird/ egg ' day day. lb. lb. 1 None 0.34 0.44 1.36 2 5% dehydrated alfalfa meal 0.33 0.45 1.42 3 10% dehydrated alfalfa meal 0.35 0.49 1.51 4 5% condensed fish solubles 0.34 0.46 1.45 5 5% condensed fish solubles + 5% dehydrated alfalfa meal 0.40 0.53 1.41 6 5% condensed fish solubles + 10% dehydrated alfalfa meal 0.33 0.52 1.66 7 5% dried brewers yeast 0.38 0.44 1.24 8 5% dried brewers yeast + 5% condensed fish solubles 0.41 0.44 1.12 14 amount of tocopherol in the eggs (Table § this instance, fertility and hatchability 1 ages were both increased when vitami added to the synthetic diet. a The degenerative changes in the eye of key embryo, obtained from hens fed die vitamin E, are of interest‘ i1; view of kn tritional deficiencies and their effect on 1 formation. Rats fed a riboflavin-deficiej develop cataracts (Day et al. 1931). Cu (1932) produced cataracts in rats by f tryptophane-deficient diet. These two su are apparently the only true primary de that were previously known to result in ,4 formation. Vitamin E has been related v lental fibroplasia, a blindness in prema man infants (Owens and Owens, 1939). , lesion which suggested retrolental fibropl reported in rat embryos due to a defic' vitamin E in the diet of the mother by l’ and-Orent-keiles (1951). ‘ ~ There was no consistent effect due t0 r; fied hatchability factor sources or to vi on egg production. However, in experi there was a marked increase in egg p I‘ when dried brewers yeast was present in In experiment 7 there was a reduction, amount of feed required to produce an e supplements of dried brewers yeast We (Table 19). The feed per egg in this ex is in close agreement with results report periment 1O (Table 13), which also c“ dried brewers yeast as one of the supple t the basal diet. ' ACKNOWLEDGMENTS This work was supported in part by: C cial Solvents Corporation, Terre Haute, I Distillation Products Industries, Rochest. York; American Dehydrators Associatio as City, Missouri; Western Condensin pany, Appleton, Wisconsin; Philip R. Pa San Pedro, California; Distillers Feed R Council, Inc., Cincinnati, Ohio; Schenley- ers, Inc., Cincinnati, Ohio; Armour and C0 Chicago 9, Illinois; Stabilized Vitamin Brooklyn, New York; Brewers Yeast Inc., Chicago, Illinois; Corn Products Y Company, Argo, Illinois; United States‘; Health Service Grant B-759, National In of Health, Bethesda, Maryland; Merck an pany, Rahway, New Jersey; Hoffman La, Inc., Nutley, New Jersey; Lederle Labo Division, American Cyanamid Company, River, New York; Bowman Feed Produc Holland, Michigan; Dow Chemical u. Freeport, Texas; and Charles A. Pfizer C0 A, Terre Haute, Indiana, v Atkinson, R. L. and J. R. Couch, 1951a. The effect of vita- ' min B12, APF concentrate, aureomycin, streptomycin, liver “L” and fish meal 0n egg production and hatch- ability of Broad BreastedBronze turkeys. Poultry Sci. 30: 905. jtkinson, R. L. and J. R. Couch, 1951b. Unpublished data. tkinson, R. L. and J. R. Couch, 195s. Unpublished data. kinson, R. L., T. M. Ferguson, J. H. Quisenberry and i J. R. Couch, 1955. Dehydrated alfalfa meal, con- densed fish solubles, distillers dried solubles and dried whey as supplements to an all vegetable protein tur- key laying diet. Poultry Sci. 34: 730-735. tkinson, R. L., T. M. Ferguson, J. H. Quisenberry and ' J. R. Couch, 1955. Vitamin E and reproduction in turkeys. J. Nutrition, 55: 387-398. vllison, E. c. and E. Orent-Keiles, 1951. Abnormalities of the eye occurring in young vitamin E deficient rats. Proc. Soc. Exp. Biol. and Med. 76: 295-297. uch, J. R., J. R. Reed, Jr., R. L. Atkinson, R. W. Per- rett, B. E. Welch and J . W. Dieckert, 1954. Distillers dried solubles for growth and hatchability. Proc. 9th Distillers Feed Conference, pp. 38-48. rtis, P. B., s. N. Hauge and H. R. Kraybill, 1932. The nutritive value of certain animal protein concentrates. J. Nutrition, 5: 503-517. P. L., W. C. Langston and C. S. O’Brien, 1939. Cata- ract and other ocular changes in vitamin G deficiency. Am. J. Ophth. 14: 1005-1009. ‘$1 'ckens, F. L., R. T. Parkhurst and c. R. Fellers, 1941. Corn distillers dried grains with solubles in poultry rations. Poultry Sci. 20: 5-36-542. eldman, G. L., 1957b. The effect of vitamin E and other " supplements on the growth and reproductive perform- ance of turkeys. M.S. Thesis, Texas A&M College, College Station, Texas. L eldman, G. L., R. L. Atkinson, B. G. Creech, T. M. Ferg- i‘ " uson, B. L. Reid and J. R. Couch, 1957a. The effect of dehydrated alfalfa meal, dried brewers yeast and condensed fish solubles on the reproductive perform- ance of turkeys. Poultry Sci. 36: 792-797. LITERATURE CITED Ferguson, T. M., R. L. Atkinson and J. R. Couch, 1954. Relationship of vitamin E to embryonic development of avian eye. Proc. Soc. Exp. Biol. and Med., 86: 868- 871. Ferguson, T. M., H. P. Vaught and J . R. Couch, 1954-1955. Unpublished data. Ferguson, T. M., H. P. Vaught, B. L. Reid and J . R. Couch, 1956. The effect of vitamin E, dehydrated alfalfa meal and condensed fish solubles upon hatchability of eggs from Broad Breasted Bronze hens maintained on litter. Poultry Sci. 35: 872-875. Ferguson, T. M., R. H. Rigdon and J. R. Couch, 1956. Cata- racts in vitamin E deficiency. An experimental study in the turkey embryo. A.M.A. Archives of Ophth. 55: 346-355. ~ 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 for the mature fowl. Poultry Sci. 32: 812-816. Jensen, L. S., 1953. Vitamin E, niacin and grass juice in turkey hen nutrition. Proc. of the Cornell Nutrition Conference, pp. 62-65. Jensen, L. S. and J . McGinnis, 1956. Quantitative require- ment of turkey breeder hens for vitamin E. Poultry Sci. 35: 1149. Nestler, R. 1a., T. c. Byerly, N. R. E1115 and H. W. Titus, " ' 1936. A new factor, not vitamin G, necessary for hatchability. Poultry Sci. 15: 67-70. Owens, W. C. and E. U. Owens, 1949. Retrolental fibro- plasia in premature infants: II Studies on the prophy- laxis of the disease; the use of alpha-tocopheryl ace- tate. Am. J. Ophth. 32: 1631-1637. Scott, M. L., and T. S. Nelson, 1955. Quantitative studies on vitamin E in turkey hatchability. Poultry Sci. 34: 1233. Stadelman, W. J., R. V. Boucherand E. W. Callenbach, 1950. The effect of vitamin D in the turkey breeder ration on egg production and hatchability and on growth and calcification of the poults. Poultry Sci. 29: 146-152. Whiteside, C. H., T. M. Ferguson and J. R. Couch, 1956. Unpublished data. 15 Location oi field research units oi the Texas Agricultural Experiment Station and cooperating agencies ORGANIZATION OPERATION Research results are carried to Texas farmers, ranchmen and homemakers by county agents and specialists of the Texas Agricultural Ex- tension Service State-wide Research .2’: i’ The Texas Agricultural Experiment Station? is the public agricultural research agency oi the State oi Texas. and is one oi ten parts oi the Texas A