FERFQRMANCE 0F LAYING MASHES COMPOSED OF CER‘E‘ME‘E MED EHGREDSHQTS A‘JAMELE 8% ?Hi UNWED STATES» GK THE UNITED ARAB REPUBLic Thesis fear fin £399:er of M. 3. MECHECSAE STATE UMVERSWY Taha Hassan Hussein. Mahmoud 196:4 THESIS This is to certify that the thesis entitled Performance of Laying Hashes Composed of Certain Feed Ingredients Available in the United States or the United Arab Republic presented by Taha Hassan Hussein Mahmoud has been accepted towards fulfillment of the requirements for Masters degree mPouitrx Science flux, (MM M/jorflro'fessor Date July 8. 1964 0-169 LIBRARY Michigan State University PERFORMANCE OF LAYING MASHES COMPOSED OF CERTAIN FEED INGREDIENTS AVAILABLE IN THE UNITED STATES OR THE UNITED ARAB REPUBLIC by Taha Hassan Hussein Mahmoud AN ABSTRACT OF A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of Poultry Science l96h ABSTRACT PERFORMANCE OF LAYING MASHES COMPOSED OF CERTAIN FEED INGREDIENTS AVAILABLE IN THE UNITED STATES OR THE UNITED ARAB REPUBLIC by Taha Hassan Hussein Mahmoud The hypothesis to be tested was whether rations composed of certain easily available and economical Egyptian feed ingredients could perform as well as typical American poultry laying rations. Three groups of lOO twenty-four-week-old pullets (four replicates of 25 small Leghorn-type pullets each) were placed for a period of six months in floor pens lO' X 8' in size with wood shaving litter, hanging feeders, and automatic waterers. Two groups of birds were fed rations containing substantial amounts of barley and cottonseed meal with and without yellow corn. A third group was provided an American-type laying ration. Comparisons were made between the three groups as to rate of lay, egg quality, fertility, hatchability, body weight gain and mortality. Neither of the Egyptian rations performed as well as the American one. The former was inferior to the latter in rate of lay, feed consumption and conversion, egg size and hatchability. All rations were similar in yolk score, shell thickness and the Haugh units. Although ferric chloride was added to rations containing cottonseed meal, a high percentage of discolored yolks was observed in fresh eggs. This percentage was greater when eggs were stored for one week, two weeks, or six weeks at 60° F. The adverse results were due to the very high gossypol level in the cottonseed meal. PERFORMANCE OF LAYING MASHES COMPOSED OF CERTAIN FEED INGREDIENTS AVAILABLE IN THE UNITED STATES OR THE UNITED ARAB REPUBLIC by Taha Hassan Hussein Mahmoud A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of Poultry Science l96h 9 a o 9 ACKNOHLEDGEMENT The author is extremely grateful to Dr. Philip J. Schaible, Professor of Poultry Science, for his guidance and Special interest in this study and his many helpful suggestions. The author also appreciated the guidance and help provided by Dr. H. C. Zindel, Chairman of the Department of Poultry Science. Dr. C. A. Lassiter, Chairman of the Department of Dairy Science, Drs. T. H. Coleman and R. K. Ringer of the Department of Poultry Science and Drs. Selma Bandemer and E. J. Benne of the Department of Biochemistry. The author appreciated the correSpondence from the University of Alexandria, United Arab Republic through his brother, Mr. Abd-El- Mohsin. Thanks also go to Michigan State University, the Department of Poultry Science and the people of Michigan for providing laboratory and farm facilities used in this study. TABLE OF CONTENTS ACKNOWLEDGEMENTS . . . . . . . . . . . TABLE OF CONTENTS . . . . . . . . . . LIST OF TABLES . . . . . . . . . . . LIST OF APPENDICES . . . . e e . o e . INTRODUCTION AND LITERATURE REVIEW . . The use of barley in chicken rations . . . Treatments to enhance the nutritional value of the barley . . . . . . . . . Laying hen rations . . O O O O O O O The use of cottonseed meal for laying hen rat‘ons O O O O O O O O O O Cottonseed meal and egg discoloration Yolk discoloration in the presence of QOSSYPOIO O O O O O O O O O O O O O O O O The main characteristics of the gossypol pink-white discoloration. . . . . . . . . Conditions which influence discoloration in eggs . . . . . . . . . . . . . . . . . Composition and nutritive value. . . . . . GENERAL EXPERIMENTAL PROCEDURES. . . . Preparation of experimental rations. . . . Egg quality determinations . Hatchability and fertility . Statistical procedures . . . EXPERIMENT I - EXPERIMENTS WITH RATIONS FOR LAYING Results and Discussion . . . . . . . . . . . . O O O O O O 0 HENS. Rate of lay . . . . . . . . . . . . . . . l2 lh I7 20 20 20 2l 2l 22 22 22 Feed consumption Egg Size . . e . Shell thickness. Yolk score . . . Haugh unit . . o Hatchability . . . . Fertility . . . . . . Embryonic mortality . Non-hatched eggs. . . Adult mortality . . . Ayerage body weight . . . . . . EXPERIMENT II - EXPERIMENTS WITH ADJUSTED Results and Discussion. Rate of lay. . . Feed consumption Feed conversion. Egg quality determinations Egg size. . . . . . . Shell thickness . . YOlk score. 0 e e e Yolk discoloration. Haugh Unlto e e e e HatChabl'ltY . e 0 e e e Fertl'lty . e e e 0 Embryonic mortality iii 0 O O O O O O O O O LAYING HEN RATIONS. £299 23 24 2% 2h 2h 25 25 25 26 26 26 Si 51 Si 51 5| 5| SI 52 52 52 53 53 53 53 Non-hatched eggs Adult mortality Body weight gain SUMMARY . . . . . . . . . . . . . . APPENDIX . . . . . . . . . . . . . . LITERATURE CITED . . . . . . . . . . iv Table ha lib 5a 5b Sc lO lla llb l2a l2b l3a LIST OF TABLES Composition of Diet A and Diet B Used in Exper'mnt I O O O O O O O O O O O O O O O O O O O 0 Composition of Basal Ration Used as a Control for ExperimntIOOOOOOOOOCHOOOOOOOOOO Calculated Nutrient Content for the Rations in ExPerImentI.................... Egg Production of Pullets Fed the Three Rations; Calculated as £995 per Hen per Day . . . . . . . . . Statistical Analysis of Differences in Egg Production Feed Consumption for the Pullets Fed the Three Rations e e o e e e e e e e e e e e e e e e e e e 0 Statistical Analysis of Differences in Feed Intake . Feed Conversion Data for the Three Rations . . . . . Weights of a Random Sample of Eggs Produced by Pullets Fed the Three Rations. . . . . . . . . . . . Shell Thickness of Eggs Produced during January by Pullets Fed the Three Rations . . . . . . . . . . Yolk Scores of Eggs Produced by Pullets Fed the Three Rations. e e e e e e e e e e e e e e e e e e 0 Number of Discolored and Normal Fresh Eggs Produced by Pullets Fed the Two Cottonseed-containing Rations e e e e e e e e e e e e e e e e e e e e e e Haugh Unit Scores for the Fresh Eggs Produced by Pullets Fed the Three Rations. . . . . . . . . . . . Hatchability of eggs Produced by Pullets Fed the Three DIets O O O O O O O O O O O O O O O O O O .0 Statistical Analysis of Differences in Hatchability. Fertility of £995 Produced by Pullets Fed the Three RationsOOOOOOOOOOOOOOOOOOOOOO Statistical Analysis of Differences in Fertility . . Embryonic Mortality of Fertile Eggs Produced by Pullets Fed the Three Diets . . . . . . . . . . . . . 28 29 30 3i 32 33 3h 35 36 37 38 39 hi 1+2 1+3 Table 13b lha ihb l5a 15b 16 l7 i8 l9a i9b 20a 20b 20c 2i 22 23 2ha Statistical Analysis of Differences in Embryonic "OftalitY e e e e o e e e e e e e e e e e e e e e e Percentages of Non-hatched Chicks Related to the Three Diets O O O O O O O O O O O O O O O O O O O 0 Statistical Analysis of Differences between Percentages of Non-hatched Chicks . . . . . . . . . Mortality during the First Experiment . . . .'. . . Statl5tlcal AnalY5l5 Of Mortality e e e e O o e e a Body Weight Gain during the First Experiment for Puiiets Fed the Three Rations . . . . . . . . . . Composition of Diet C and Diet D Used in Experiment 11 o o o e e e e o e o e e e e o e e o 0 Calculated Nutrient Content for the Rations Used in ExPeriment II . . . . . . . . . . . . . . . . . . . Egg Production of Puiiets Fed the Three Rations; Calculated as Eggs per hen per day. . . . . . . . . Statistical Analysis of Differences in Egg PFOdUCthfl e e e e e e e o e e e e e e e e e e e 0 Feed Consumption for the Puiiets Fed the Three Rations O O O O O O O O O O O O O O O O O O O 0 Statistical Analysis of Differences in Feed Intake 0 O O O O O O O O O O O 0 O O O O O O 0 Feed Conversion Data for the Three Rations. . . . . Heights of a Random Sample of Eggs Produced by Puiiets Fed the Three Rations . . . . . . . . . . . Sheli Thickness of Eggs Produced during April by Puiiets Fed the Three Rations . . . . . . . . . . . Yolk Scores of Eggs Produced by Puiiets Fed the Three Rations O O O O O O O O O O O O O O O C O O 0 Number of Discolored and Normal Fresh Eggs Produced by Puiiets Fed the Two Cottonseed-containing Rations O O O O O O O O O O O O O O O O O O 0 vi “7 5 50 5h 55 56 57 58 59 60 6i 62 63 Table Zhb 21m 24d 25 26a 26b 27a 27b 28a 28b 293 29b 30a 30b 3i Number of Discolored and Normal Stored Eggs for One Week at 60° F. Produced by Puiiets Fed the Two Cottonseed-containing Rations . . . . . . . . . . . Number of Discolored and Normal Eggs Stored for Two Weeks at GOP F. Produced by Puiiets Fed the Two Cottonseed-containing Rations . . . . . . . . . Number of Discolored and Normal Eggs Stored for Six weeks at 6OP F. Produced by Puiiets Fed the Two Cottonseed-containing Rations . . . . . . . . . Haugh Unit Scores for the Fresh Eggs Produced by Puiiets Fed the Three Rations . . . . . . . . . . . Hatchability of Eggs Produced by Puiiets Fed the Three Diets O O O O O O O O O O O O O O O O O O O 0 Statistical Analysis of Differences in Hatchability Fertility of Eggs Produced by Puiiets Fed the Three Rations O O O O O O O O O O O O O O 0 O O O O O O 0 Statistical Analysis of Differences in Fertility. . Embryonic Mortality of Fertile Eggs Produced by Puiiets Fed the Three Diets . . . . . . . . . . . . Statistical Analysis of Differences in Embryonic MortalitYOOOOOOOOOO00.0.0.0... Percentages Non-hatched Chicks Related to the ThreeoietsOOOOOOOOOOOO00...... Statistical Analysis of Differences between the Percentages of Non-hatched Chicks . . . . . . . . . Mortality during the Second Experiment. . . . . . . Statistical Analysis of Mortality . . . . . . . . . Body Weight Gain during the Second Experiment for Puiiets Fed the Three Diets . . . . . . . . . . . . vii 66 67 68 69 7o 7i 72 73 7h 75 76 77 78 79 Table LIST OF APPENDICES Prices of Yellow Corn and Barley in Michigan and the United Arab Republic . . . . . . . . . . . Composition of Vitamin-Trace Mineral Mix . . . . . Labeling of Cottonseed Meal Used in the Experiment viii Page ‘81 82 82a INTRODUCTION AND LITERATURE REVIEW Egypt's poultry industry is based primarily on small flocks owned by general farmers (Asker, l93h, El-Ibiary, l955). Flocks are mostly fowl. ranging in size from six to twenty birds and weigh three to four pounds each. There are no well-defined breeds. They are classified locally into four groups, Baladl, Dandarawi, Fayoumi and Eskandaranl. The latter is new and is a cross of Fayoumi with Rhode Island Red. White Leghorn or Barred Rock. The chickens are usually fed table scraps and scavenge for other food in the village streets (Ei-Ibiary, l955). There is very little expenditure for commercial feed and no investment in housing. Possibly left- overs from the soldiers' meals (Army, Navy and Air Force) could be processed into a byproduct which might be added to commercial poultry feeds. Although the poultry industry in the United Arab Republic is presently many years behind that of the United States of America, it wms at one time in the forefront. Foreign visitors during the Middle Ages, or thereafter, were fascinated by the new'method the Egyptians used to get their chicks. Early writers stated that "all its details are kept most religiously secret (Bey. i927; Ei-Ibiary, l9h7, l955; Reid and Dardiry, i953). but that the attendant used to sit on the eggs until hatching. Thus, artificial incubation has been known for a very long time in Egypt. Until about twenty-five years ago. eggs were a chief export of Egypt* (Ragab and Assem. l953). They lost their export market. chiefly because of their small size and lack of unifonmity. *The peak years for exportation of eggs were l92h and i932 when more than 200 million eggs were sold. The average protein intake in Egypt is only ll grams per person per day, much below'the minimum requirement of 33 grams suggested by the U. 5. National Research Council (Phillips, l95l). It would be desirable to increase animal production in Egypt, but there is a shortage of roughages and concentrates for feeding (Ahmed and Badr, l953). One hard fact governs Egypt's agricultural situation; of its total area of about 2A7 million acres. almost 97 Percent is desert or otherwise unsuitable for agriculture. Each year the number of the Egyptians increases by over a half million, while the arable expands only slightly (Robbins. l95h). An expanded poultry industry might contribute importantly to the nutritional well-being of the Egyptians but some of the feeds used for. poultry, such as barley, are used directly in the human diet. Barley as a food for man dates back to 5,000 B.C. and before. Until the sixteenth century it was the chief grain for breadmaking in Europe and the Mediterranean region; then it was supplanted by wheat and potatoes. Even today. barley is widely used because it can be grown in a greater variety of climates than corn, wheat, or oats. In the United Arab Republic the Beduins (people who live in the desert) still make their bread from barley. In America, barley is used mainly as an animal food and for manu- facture of malt. In Canada and the U. S. Pacific coast, barley is grown extensively and used in poultry and swine feeds (Ewing, l9h7). It is used in about the same way as oats and may replace to a large extent oats, wheat, wheat byproducts. or corn in the ration. The price of barley in the United.Arab Republic is substantially lower than that of corn* (Ei-Ibiary, l955). This fact leads to the iThis is true also after nutritive adjustment according toTFFaps (l9h6). (See Appendix) extensive use of large quantities of barley in poultry diets. A major question from a nutritional standpoint is how'much barley can be used efficiently in these poultry diets. The difference in nutritional value of corn and barley for poultry has been recognized for decades. A quantitative measure of this difference was given by Fraps (l9h6) in terms of productive energy content of the two grains. Based on his results with chicks. barley has only 70 percent of the feeding value of corn. To overcome this lack of energy in barley. added fat is used in United States diets, but oils and fats cannot be economically used in the United Arab Republic. The methionine content of barley is approximately one-half to two-, thirds that of corn (Arscott et al.. l959). Like oats. it contains only an insignificant trace of vitamin A activity (Ewing. l9h7). Wagstaff _e_l_:__a_l. (i959) stated that chicks fed diets containing barley hulls or regular barley showed typical biotin deficiency symptoms. These products are more deficient in biotin than corn. Furthermore, birds not accustomed to barley do not consume it readily (Ewing, l9h7). Recent studies show quite convincingly that barley may be made equal to corn in nutritional value for poultry either by treating with water or feeding certain enzymes with it in the ration. If similar methods improved the nutritional value of barley for other monogastric animals, including man, future years may see an upswing in the use of the barley as human food in areas where it can be grown economically. The expansion of the poultry industry in recent years has also resulted in greater demands for protein concentrates of both animal and vegetable origin. Extensive studies have been made relative to the use of cottonseed meal, Egyptian feedstuff in ample supply. in rations for laying hens. The majority of these studies indicate that for egg pro- duction, cottonseed meal may replace a part of the animal protein. Unfortunately, a characteristic darkening of the yolk occurs upon cold storage of the eggs of hens fed this feedstuff which renders them unacceptable for sale. Consequently. the poultrymen have not been able to utilize cottonseed meal to any great extent in laying rations. Con- flicting reports on the effects of cottonseed meal on the internal quality of stored eggs indicates that marked differences occur in the cottonseed meal samples with reSpect to botanical, climatic, geographical and processing plant histories. It should be emphasized that "discolored yolks caused by hens consuming large amounts of cottonseed meal are not Spoiled or unwholesome but are sweet. clean and perfectly edible" (Ewing. 19W) . Cottonseed meal processed so that it is very low in free gossypol can be fed to hens without injuring the storage quality of the eggs. Halloran and Cavanaugh (l960) state that cottonseed meals can be produced commercially with an ASU** level of 0.3 or less. Of course, the eggs produced should be stored at 33° F. Glandless cottonseed meal. degossy- poiized cottonseed meal or oil dipping and spraying of eggs might permit a much greater use of cottonseed meals in poultry rations in Egypt. The hypotheses to be tested is whether easily available and economical Egyptian feed ingredientsiwill perform as well as American ones in poultry laying rations. The research reported herein will be of greatest interest to owners of big flock farms, such as the government, the amateur's Poultry Breeds Association. and educational institutions. *tlvaiiable gossypol units The Use of Barley in Chicken Rations Arscott.g£.gl. (l959) showed that barley used up to 25 percent of the grain component (l3 to i5.3$% of the total ration) in a high efficiency broiler ration exerted no significant adverse effect on growth of chicks; the growth was depressed when 50 to lOO percent barley was included in the ration. [As the barley content of the rations increased, body pigmentation decreased. Since the hull of the barley is responsible for its lower nutritional value, a new hulless variety was studied by Anderson‘gtugl. (l96l). He reported that the rate of gain and gain/feed ratio of chicks fed a ration based on yellow corn averaged i2 percent greater than that of chicks fed a ration based on the hulless barley. Rations based on hulless barley were not superior to those based on regular barley. Treatment to Enhance the Nutritional Value of Bariey--Dobson and Anderson (l958), Arscott (l958), Fry £31. (i957). Fry 25:1. (l958). Wharton‘gt'gl. (l958). Berg (i959), and Arscott (l960) obtained marked improvement in growth of chicks when barley was treated with water prior to mixing or when certain enzymes were added with the barley to the feed. Willingham.gt‘gl. (l960) reported that Western barley is improved when it is water treated, but other investigators (Gerry, l959) have failed in this regard. The generally accepted hypotheses are that the chick does not . produce enzymes required for the digestibility of barley as it contains an inhibitor of tryptic activity (Fry 3331., l957). Arscott 35;], (l960) suggested that water treatment of barley removed the inhibitor and permitted tryptic activity to take place nonmally. Willingham'gt.2l. (l959) indicated that a highly significant im- provement in growth and feed utilization occurred when diets containing barley were supplemented with enzymes from bacterial or fungal sources and malted barley. However, crystalline alpha amylase was completely inactive. Barley autoclaved* wet was not improved by dietary enzyme supplementation. Ten percent of malt was generally effective in improving growth. Rose and Arscott (l962) reported that when corn constituted one- half of the ground grain of the barley type ration. results were comparable to supplementing a barley ration with an amylolltic enzyme. An attempt was made by Arscott (l963) to reduce the amount of corn necessary to elicit a reSponse to barley similar to that obtained from an amylolltic enzyme supplement. Replacing l/8 to i/h barley in the ration with corn proved equally as effective as an amylolltic supplement, when data on body weight, feed conversion, and accumulated droppings are considered. Broilers fed rations containing corn as the sole grain component still proved superior to rations containing barley, amylolltic enzyme or corn- barley combinations. Rose‘gt‘gl. (l962) stated that at least 2.5 pounds of an amylolytic enzyme supplement** per ton of feed were necessary for a pelleted barley ration containing three percent fat to elicit responses comparable to those obtained from a mash ration containing corn. All levels (l.25, 3.5 and 5.0 pounds per ton) of the enzyme supplement markedly iamroved the "sticky droppings“ condition. Where corn replaced one-half the barley in a pelleted ration, perfonmance was comparable to the corn mash ration. Anderson_g§.gl. (l96l) reported that maintaining the temperature * Processed under l5 lbs. pressure. **Dawes Laboratory, Chicago, Ill. of the grain during water soaking at 85-90? C. followed by drying at 6OP C. made the soaking procedure effective, but a higher temperature (80 - l20p C.) did not. Thomas'gtugl. (l960) reported that water treated barley contained many more bacteria than untreated barley. even though drying temperature was 70? C. Thomas_g£fl21. (l96l) indicated that autoclaved barley was significantly improved for chicks by water treatment. Therefore. enzymes endogenous to barley were not necessary for improvement. nor is its physical effect from water treatment. It was concluded that enzymes or other factors produced by microorganisms bring about the nutritional im- provement of barley by water treatment. 0n the other hand, Leong‘gtlgl. (l962) reported that the addition of a fungal enzyme supplement increased the metabolizable energy of Western pearled barley and regular barley 23.8 and lh.5 percent, reSpectively. whereas water treatment resulted in increases of l6.l and 2#.6 percent in metabolizable energy. Laying Hen Rations--Berg 23,21. (l9S9) have shown that under the climatic conditions of western Washington, hens would lay equally well on corn and barley diets. However. barley was only 82.7 percent as efficient as corn for the production of eggs. Hens fed barley rations voided a larger volume of wetter droppings than hens fed corn diets. The addition of funial and bacterial enzymes barley rations did not affect the rate of lay, amount of feed per dozen eggs, body weight gains. hatchability of fertile eggs. egg quality or yolk color. The effect of water treatment was the same as enzyme addition, except that there was a small reduction in food required per dozen eggs. The fungal enzyme preparation in the feed decreased moisture content of the litter and improved growth rate and feed efficiency in young chicks fed barley rations. The results obtained might be related to the age of pullets (29 weeks) and the fact they were in full lay (Berg, l96l). The addition of enzyme supplement to the diet containing barley increased growth rate up to eight weeks of age but had no effect during the period between 8 to 2i weeks. It increased body weight gains during the laying period and also improved the condition of the litter on which the birds were reared to 2i weeks of age. No significant improvement in rate of lay or feed efficiency was obtained by addition of enzymes at any of the time intervals studied. Likewise, no benefit from enzyme supplementation was obtained for egg weight, Haugh units*, specific gravity of eggs, hatchablllty of fertile eggs or livability of birds. Nelson (l958) reported that the addition of enzymes or water treating the barley improved egg production and feed efficiency over the barley-fed group. The added enzyme gave a significant increase in hatchablllty over the barley and soaked barley diets. There was little difference in egg size, egg quality or weight gains between the different diets. The Use of Cottonseed Meal for Laying Men Rations The cotton plant produces more food for man and feed for livestock than it does of the fiber that bears its name**. Cottonseed Meal and Egg Discoloration--Robert and Rice (l89l)*** noted that egg yolks were discolored in cold storage in eggs laid by hens fed rations containing cottonseed meal. However, no real attempt was made to determine the cause or correction of this discoloration until about * Haugh units are the albumen height adjusted according to the weight of the egg. (For details see Haugh. R. R.. l937. The Haugh unit for measuring egg quality. U. 5. Egg 5 Poultry Mag. #3: 552-555, 572-573.) ** National Cottonseed Products Ass'n., Inc. Cottonseed‘ggg.;£s Products. ***Cited from Heywang at _a_l_., l963. l930. By then, it became recognized that two discolorations were in evidence. Cottonseed and its byproducts are implicated in two ways: i. Gossypol present in cottonseed and their products produced olive or chocolate-brown yolks described by Schaibie‘g£‘_l. (l93h) and Swenson‘gt'gl. (19h2). 2. Salmon-colored yolks and pink whites associated with cottonseed meal feeding as reported by Sherwood (l928), (l93l). Yolk Discoloration in the Presence of Gossypol: l. Pure gossypol (Lorenz, l939) 2. Cottonseed meal processed by different methods (Meywang at 1955; Evans 55, 31.. 1957) 3. Cottonseed oil (Sherwood, l93l; Evans'££.gl., i957) h. Crude cottonseed oil (Sherwood, l93l; Evans.g£.gl., l957) 5. Cottonseed still bottoms (Pepper‘gt'gl., l962) 6. Ether-soluble soapstock (Evans 35 31., l957) 7. Raw cottonseed (Lorenz, l939; Heywang'gt‘gl., l9h9, l95h) 8. Ether extract of cottonseed or meal (Sherwood, l93l; Lorenz and Elmquist, i93h; Schaible‘gtlgl., l93h) 9. Partially refined cottonseed oil (Sherwood, l93l) l0. Cottonseed pigment glands (Heywang‘g£.gl., l95h) The data reported by Heywang‘gt‘gl. (l955) indicated that cottonseed meal rations of equal gossypol content would not produce equal incidence and intensity of yolk discoloration. The amount of cottonseed meal that may be fed safely to simple-stomach animals and young ruminants was thought to be dependent upon the level of free gossypol in the meal. Although gossypol in the bound form is not considered toxic, it reduces the nutritive quality of the meals. Developments in processing cotton- seed have reduced the hazards from feeding and improved the quality of l0 the oil, but the undesirable effects of gossypol have not been eliminated entirely. .As mentioned before, earlier workers presumed that coloration develops because of the "free“ gossypol present in the meals (Heywang £531., l955; Heywang 5531., l95‘i; Schaible 5.5;]... l93li; Swenson £331., l9h2). Apparently, cottonseed meals cannot be graded for laying rations on the basis of their "free" gossypol contents, however, since the correlation between intensity of coloration in the yolks and free gossypol content of the meals fed is poor (Framptonig£.gl., l96l). A.method for grading cottonseed meals for laying rations proposed by Grau (l95h, l955) depends upon a greater concentration in the yolks of some acetone-hexane soluble material. A cottonseed meal having an AGU* of 0.30 or less was reported by Grau to be suitable for laying rations in the amount up to ten percent of the total ration. The AGU values were almost invariably positive (Frampton g£_§fl,, l96l). Considerable improvement may be effected by developing varieties of cotton in which the gossypol content is less than 0.50 percent which level yields meals containing less than 0.85 percent total gossypol (Hamilton, l958). Cotton geneticists have des- cribed genes which enable the gossypol content of the seed to be reduced without altering desirable characteristics of the cotton plant. Two geneS. glz and gl3 were found reSponsible for glandlessness (absence of pigment glands) in the cotton plant. Gossypium hirsutum‘L. glandless cottonseed, which is associated with absence of gossypol and other pig- ments, promises to be of economic value to cottonseed crushing industry (McMichel, 1960). *AGU - (AFEOD - A'hSO) - ANROO - A9450) X l00 Percent of the material tested in the ration A' refers to the absorbance of the extract from cottonseed meal-produced eggs; A” refers to the absorbance of the extract from the control eggs. The new cottonseed meals could be incorporated in the broiler rations to supply twenty percent protein with no excess of the 0.2 percent dietary free gossypol levels, above which Heywang and Bird found White Leghorn and New Hampshire chicks affected. Laying hens fed essentially gossypol-free, low-oil, glandless cotton- seed meals produced eggs which maintained excellent quality during several months cold storage (Phelps, i962). Cottonseed meal which had been sub- jected to solvent treatment during processing, and presumably contained less oil and cyclopropene fatty acids, produced less dark yolk discoloration when compared to meals processed by pressing operations. Phelps‘gt'gl. (1963) stated that the dark olive discoloration is due to a combination of yolk gossypol with yolk iron. Two subsequent studies (Kemmerer'gt‘gl., l962, l963) indicate that partially refined cottonseed oil, when fed at a relatively high level, will act like‘§. foetida oil in enhancing gossypol-induced yolk discoloration. The intensity of yolk discoloration after three months storage of eggs from hens fed 6 mg/day of gossypol and 2.5 percent cottonseed oil was similar to that observed in three-month-stored eggs from hens fed 50 mg/day of sterculia foetida oil plus six mg of gossypol daily (Kemmerer‘gt‘gl., l96l). The estimated daily intake of cyclopropene fatty acids for the hens receiving the 2.5 percent cottonseed oil ration would be from l4 to 18 mg‘g. foetida oil fed at the highest level of 50 mg/day. More recent data (Kemmerer‘gt'gl., l963) obtained by feeding 3 mg/day of gossypol with O.l percent cotton- seed oil suggests that practical levels of low residual oil cottonseed meals may not significantly enhance yolk color in stored eggs. l2 The Iain Characteristics of the Gossypol--Glands containing various pigments, including gossypol, are common to all Species of the genus Gossypium. The first isolation of a yellow'pigment was recorded by an English chemist (Anon, l886; Bailey, l9h8). Several years later, a Polish chemist, Harchlewski, purified the pigment and demonstrated the polyphenolic character of the compound and named it gossypol, from gossyp (ium phen) ol, to indicate both its origin in cottonseed and its chemical nature. (For more details read Bailey, l9h8). Though gossypol is light yellow, it is usually singled out for special emphasis because of its toxic properties. Toxicity due to gossypol limits the use of cottonseed meal as a feed unless the gossypol ls inactiv- ated or removed in processing the seed. Gossypol is soluble in many substances as cold dioxane, diethylene glycol, methanol, ethanol, etc. When heated, gossypol melts with decomposition. Since gossypol is unstable in solution at room temperature, it may be concluded that it would decouh pose or rearrange at higher temperatures. Gossypol reacts as a strong dibasic acid, forming neutral disodium and dipotassium salts which are colored a deeper yellow than the free compound. The iron which is normally bound by the egg yolk proteins is released during storage and this iron combines with gossypol to form the product which is responsible for the dark color of the egg yolks (Swenson, i9h2). Comprehensive details about the gossypol structure, its functional groups, and its estimation are given by Curtis (l938), Bailey (l9h8), Crau'g£.gl. (l95h, l955), Uoronick and Grau (l955). Pink-White Discoloration--Sherwood (l928, l93l) described, many years ago, the pink-white discoloration of stored eggs from hens fed cottonseed meal. If most of the oil was extracted from the meal, however, 13 the egg discoloration was reduced greatly. Similarly, the feeding of refined cottonseed salad oil resulted in eggs which did not discolor when stored. Evans'g£.gl. (1957) reported that feeding of either crude cotton- seed oil or cottonseed meal to hens resulted in production of pink whites. Since raw cottonseed contains both gossypol and the Halphen* substances, it is believed that this type of discoloration is the resultant of the action of both substances. Evans £3 21. (l960) stated that stored eggs from hens fed crude cottonseed oil developed pink whites and large salmon or large brown thick translucent curdled yolks. No egg discoloration was observed when crude cottonseed oils, heated at 200° C. for eight hours or at 2H0? C. for one hour, were fed. The recent reports of Kemmerer‘gt‘gl. (l960) and Frampton'_£._l. (l96l) show that the dark discoloration caused by gossypol and appearing in yolk of stored eggs from layers fed cottonseed meal is accompanied by an increase in the pH and is enhanced by another component of cottonseed meal (halphanic acid), which also causes pink-white and salmon-colored yolks. Evans._£'gl. (l962) reported that heated, Halphen-test-positive cottonseed oils resulted in normal colored eggs when fed to laying hens. Furthermore, a Halphen negative fraction of cottonseed oll fatty acids not precipitated by urea produced pink-white discoloration when fed to laying hens, whereas the remaining fraction precipitated by urea gave a positive Halphen test but resulted in normal colored eggs when fed. The pink color was reported by Hasson 21:. 11. (I957) and by Shenstone and Vickery (l959) to be caused by prOpene ring fatty acid. Further information has been accumulated concerning the molecular structure *The presence of pink or red color indicates a positive Halphen reaction. (See Bailey, l96l) lh required for producing both the Halphen test and pink whites in eggs by the preparation of derivatives of sterculic acid (Nunn, 1952; Nordby ‘gt'gl., 1962). Schaible and his associates concluded that the cause of pink egg white discoloration can be attributed to a combination of conalbumin, an egg white protein, with yolk iron (Bandemer and Schaible, 1996; Bandemer‘gt‘gl., 1996; Schaible and Bandemer, 1996a, 1946b; Schaible ._£.gl., 1996). Azari* and Feeney (1961) reported that conalbumin binds two ferric ions per molecule of conalbumin and that the complex is salmon or pink colored and relatively resistant to chemical and physical treat- ments. Evans gtugl. (I959, 1959) reported that several egg white proteins of eggs from hens fed 2.5 percent crude cottonseed oil migrated to the. yolk during six months cold storage. Ovalbumin, conalbumin, and lysozyme migrated to the yolk but not ovomucoid or ovoglobulin. Livetin migrated from the yolk to the white. Schaible and Bandemer (1996a) had shown previously that iron migrated from the yolk to the white under similar experimental conditions. Pink discoloration of eggs has been reported to occur when non- cottonseed rations were fed but a longer storage period is usually required (Thompson, 1939; Schaible ££.El-9 1996). This indicates that the onset of the pink-white disorder is a highly accelerated manifestation of some change which may occur in normal eggs. Conditions Hhich Influence Discoloration in Eggs-~These could be the main factors which influence discoloration in eggs laid by hens fed cottonseed meal. 1. Types of Cottonseed Heals -- To obtain successful results, cottonseed meal for laying rations must be low in both free gossypol, *Cited from Pehlps £5 21,, 1963 15 or A60 and residual lipids. At the ten percent level in the ration, meals should not exceed .Oh percent free gossypol and 1.0 percent residual lipids (measured by extraction with diethyl ether). By selecting lowbgossypol or solvent-processed cottonseed meals, poultrymen can successfully employ cottonseed meal in laying rations. 2. Methods of Processing -- The cottonseed meal produced by hydraulic or expeller methods vary from that produced by solvent-process methods in fat and gossypol content. Evans‘g£.gl. (1960) stated that no egg discoloration was observed when crude cottonseed oils heated at 2009 C. for eight hours or at 290p C. for one hour were fed. 3. Feeding Period -- After a feeding period of only four days, pink- white discoloration with a positive Halphen test in stored eggs may develop if sufficient cyclopropenoid fatty acid-containing materials are fed (Lorenz, 1939). On the other hand, Heywang'g£.g1. (1959, 1963) reported that a ten-day period is necessary after removal of cottonseed from the diet of layers to completely eliminate discolored eggs, 9. Storage Period -- Heywang gtflgl. (1955) reported that longer storage periods result in a higher incidence of pink discoloration. Deutschman g£_£fl, (1961) also reported an increased incidence of pink-whites as the storage period was increased. 5. Storage Temperature -- Heywang‘gtlgl. (1962) reported that treated eggs showed no yolk discolorations when the storage temperature was 35° F., but not when it was 50 - 55° F. Kemmerer‘gt‘gl, (1962) found that the simultaneous feeding of crude cottonseed oil with crystal- line gossypol to laying hens intensified the discoloration caused by gossypol in eggs stored for one to three months at 35° F. 16 Pepper‘gt‘gl. (1962) reported that the storage time required to develop pink-white discoloration decreased when storage temperature was increased from -1° C. to 16° C. t'gl. (1932) reported that eggs from hens fed large Thompson amounts of cottonseed meal did not discolor when stored in a frozen condition, either whole or with white and yolk separated. Schaible _£_gl, (1996) stored eggs from hens fed either twenty percent cotton- seed meal or five percent crude, cold pressed, expeller cottonseed oil in a frozen state, with whites and yolks separated, for nine months. Whole eggs showed a pink color after nine months of storage at OF C. or six months at 99 C. but the separated yolks and whites did not discolor. 6. Treatment of Cottonseed Meal -- The treatment of cottonseed meal with $02 appeared to destroy both the pink white discoloring capacity and the olive yolk effect of the cottonseed meal (Deutschman t al., 1961). Card (1952) stated that 0.5 percent ferric chloride or ferrous sulfate fed along with cottonseed meal will combine with the gossypol and prevent its absorption from the intestinal tract. Without this precaution, no more than five percent cottonseed meal may be safely fed to laying hens. Heywang (1957) stated that 0.25 and 0.5 percent FeSOu inactivated some of the free gossypol, but not all of it. Cottonseed meal treated with HCl caused no discolored whites or yolks in eggs stored for six months (Deutschman _£._l., l96l). Eggs from hens fed cottonseed meal treated with $02 gas prior to feeding failed to discolor when exposed to an atmOSphere of NH3 for three hours. 17 t a1. (1961) stored eggs 7. Treatment of Stored Eggs -- Deutschman from hens fed twenty percent cottonseed oil meal under air, oxygen, carbon dioxide or nitrogen and obtained no differences in the incidence of pink whites. Eggs showing the pink whites yielded pH values of 8.26 and 8.32, reSpectively. Heywang gt‘gl. (1962) reported that dipping fresh eggs in oil, apparently prevented the formation of dark yolk discoloration when dietary level of free gossypol was less than .008 percent. Spraying was effective at the .001 and .002 percent levels of free gossypol when the storage temperature was 35° F. Composition and Nutritive Value Cottonseed meal of the usual grade contains #1 percent protein or. more, thus having nearly as much as soybean oil meal. The nutritive quality of cottonseed meal is impaired when it is heated (Condon‘gt'gl., l95h), partly because of the destruction of a portion of the lysine in the proteins (Conkerton e_t__a_]_., 1957; Martinez _e_t§_1_., 1958). The mechanisms of this destruction is not known (Martinez 25.21., 1961). It is possible that both carbohydrates and gossypol are involved, as amino acids are deaminated and decarboxylated when heated in the presence of either of these two carbonyl or potential carbonyl-containing substances (Frampton ‘g£.gl., 1958). A part of the impairment may also be due to the unavaila- bility of lysine in the meal to the animal because of the reaction l_.situ of gossypol, sugar, or other constituents with the free epsilon groups of lysine in the proteins (Conkerton‘gt‘gl., 1959). Hartinez‘gt‘gl. (1961) reported that the reduction in nutritive value in processed cottonseed is due not only to lysine destruction but to lysine binding as well. 18 Approximately one-fifth of the lysine content of the seed may be destroyed in conventional commercial operations. The quantity of lysine destroyed is dependent upon the "severity" of the processing, and not upon the processing used (Martinez and Frampton, 1958). On the other hand, Zablan gtflgl. (1963) stated that, while the estimated metabolizable energy from the approximate analysis of a par- ticular sample of cottonseed oil meal was 1260, it was found to be only 920 calories per pound as determined biologically. The quality of the cottonseed meal apparently cannot be evaluated satisfactorily by quantities of various amino acids present (Miner and Watts, 1961). Phelps (1962) reported that presently manufactured cottonseed meals contain gossypol, and also have a portion of the lysine bound in a form not utilizable by nonruminant animals. The lysine is believed to be bound to non-gossypol components of the seed, as well as to gossypol. The binding occurs during production. Improved response following lysine supplementation of cottonseed meal-containing rations afford evidence of the need for additional lysine. Further evidence of the need for lysine has been shown by blood amino acid studies. Studies have demonstrated the advantage of mild heat treatment during processing, and the benefit of solvents which remove most of the gossypol. However, cottonseed meal does not furnish protein of high quality for poultry, partly because of its low lysine content and also because of toxicity of the gossypol. 0n the other hand, Halloran and Cavanaugh (1960) stated that cottonseed meals could be produced commercially which would have an AGU level of 0.3 percent and less. At 33° F. storage, they found no objectionably colored yolks after three and six months' storage. l9 Glandless cottonseed offers outstanding opportunity for the cotton- seed industry to improve the quality of its products through breeding. Even glandless meals had a need for additional lysine when furnished the main protein supplement for chick rations. Results of glandless meals, however, were superior to those with glanded cottonseed meals. Cottonseed meal produced by the hydraulic or expeller methods has five percent more fat. The fat content was somewhat higher prior to new processing methods which made possible more complete removal of the oil. Solvent-process cottonseed meal is much lower in fat. Hydraulic or expeller process cottonseed meal has about 1.91 percent fat and solvent- process cottonseed meal has about 1.91 percent fat. High-grade cottonseed meal supplies slightly less total digestible nutrients than linseed meal. The solvent-process meal is somewhat lower than the expeller-process or hydraulic-process meal because of the lower fat content. Cottonseed meal is one of the richest feeds in phOSphorus, containing about 1.0 percent or more. It has only about 0.2 percent calcium. Like other seed products, cottonseed meal lacks vitamin D and has little or no carotene. Ramey (1962) concluded that increasing the polyunsaturate to saturate ratio can be done through increasing the amount of linolenic acid. GENERAL EXPERIMENTAL PROCEDURE Preparation of Experimental Rations To compare ingredients, composition tables and chemical analyses were used. Some of the ingredients used were substitutes for other ingredients. For example, the Egyptian Berseem* (Trifolium alexanderinum) was replaced by alfalfa meal, 17%.protein. The calculated protein analysis of the former ranges from 16 to 18 percent. The use of five percent alfalfa was to obtain acceptable yolk pigmentation. Fine wheat bran was replaced by coarse wheat bran. No attempts were made to water treat the barley, or use enzyme supplementation in the feed. Diets were prepared monthly. When the ferric chloride was used during the second phase of the experiment, efforts were made to obtain thorough distribution of the chemical in the cottonseed meal prior to mixing with other ingredients. Feed was weighed back on the first day of the month during the whole experimental period. Egganlity_0eterm1nations Birds were trapnested five days a week and collected daily. Each month a week's eggs were broken out and examined for egg quality factors as influenced by feed treatments. Albumin quality was determined in Haugh units. A micrometer was used to measure shell thickness. Yolk color score was determined by comparison with the Heiman and Carver roter** device. The ammonia atmOSphere test was given by Schaible (193h) and was employed for predicting yolk discoloration. * Morrison, 1962, p. 574. ** U. S. Egg and Poultry Magazine XLi: 8h0, Aug.. 1935. 20 21 All egg quality measurements, except yolk discoloration, were determined on fresh eggs. However, to obtain potentialities for dis- coloration, a random sample of eggs were stored at 50° F. and tested after one, two, and six weeks. Hatchability and Fertility Hatchability was calculated as the percentage of sound chicks that hatched from fertile eggs. Malformed chicks were not considered sound chicks but were included with non-hatched chicks. Dead embryos and the non-hatched chicks were calculated as a percentage of fertile eggs. Birds were weighed individually at the beginning and end of the experimental periods in order to ascertain any gain of body weight. Statistical Procedures Certain data from the experiments were subjected to statistical analysis, usually by the analysis of variance. The "t" test, and standards of deviation were also used. Individual as well as group differences for such characteristics as weights of the birds, weights of the eggs, etc. were analyzed statistically for significance at the 5 and 1 percent levels of probability. When the fertility reached zero percent in pen number 3 during the first experiment, adjustments outlined by Cochran and Cox (1957) were made in an effort to overcome this problem. Tables of hatchablllty, fertility, embryonic mortality and percent non-hatched chicks were converted as shown by Snedecor (1950). EXPERIMENT I - EXPERIMENTS WITH RATIONS FOR LAYING HENS Three groups of 100 small, twenty-four-week-old Leghorn pullets (9 replicates of 25 birds each) were placed in floor pens 10' X 8' in size. Pens and birds were chosen at random. Wood shaving litter, hanging feeders, and automatic waterers were used in all pens. Two groups were fed cottonseed meal and barley in rations with and without yellow corn. The third, control group, was fed an egg mash patterned after present-day commercial feeds. Compositions of diets A and B are given in Table 1 and the control in Table 2. Calculated nutrient contents of the three rations are shown in Table 3. The weights of birds at the beginning of the experiment are shown. in Table 16. A whole week's egg yield was tested monthly for hatcha- bliity and fertility. Incubated eggs were candied for fertility and dead embryos on the eighteenth day of incubation. Results and Discussion of Experiment I 1. Rate of Lay - From Table ha, it is obvious that the control diet was superior to other diets in promoting egg production. Pullets on the control diet laid almost twice as many eggs as those on diet A, and about 2.5 times those on diet 8. According to Berg 35 21. (1959, 1960), there is no ad- verse effect on egg production when pullets are fed rations con- taining untreated barley. But Heywang and Bird (l95h) found that free gossypol depressed egg production and feed consumption when its level in the diet reached .016 percent. According to their results, the average number of eggs per hen per day decreased to .h2 when the pullets were fed a diet containing a level of 0.025 percent free 22 23 gossypol. Although the free gossypol content in the cottonseed meals used in this experiment is not known, it is evident from the low rate of lay that it is very high. This is further substantiated by the fact that all of the eggs tested had discolored yolks when freshly laid (Table 9). In Table hb, the F ratio indicates that these differences were highly significant (P (.01). There were also significant seasonal differences in egg production with all feed treatments with re5pect to different months during the experimental period but no interation. 2. Feed Consumption - The drop in egg production was accompanied by a decrease of feed consumption for groups fed cottonseed meal (Tables 5a, 5c). From Table 5a, the average total feed consumption per pullet was from about 10 to uh percent lower for dietsiA and 8, compared to groups fed the control ration. These differences were found to be highly significant. Heywang and Bird (195h) reported that the average total feed consumption/pullet/day reached 66.7 grams when birds were fed a diet high in gossypol level (0.25% free gossypol) compared to 88.9 grams for the control with a de- crease of about 25 percent. Feed intake between the four different months of the experimental period, was found not to be significant and there was no interaction (Table 5c). The feed conversion was a third to a quarter as good on diet A than the control ration and only half as good on diet B as the control diet (Table 5a). According to these results, these particular ration formu- lations could not be used for a commercial feed. 2h 3. Egg Quality Determinations - Eggs produced by pullets fed diet A or B were smaller than those produced by birds fed the control ration (Table 6). The calculated standard deviation for each pen shows that there were some variations in egg sizes within the pen. The ”t” test indicated that these differences between diets were significant (P (.01 and P (.05). Heywang and Bird (1950) repdrted that free gossypol at the .02“ percent level had an adverse effect on egg weight, this effect was greater at the .036 percent level. It is evident that the small size of eggs produced by pullets on diets A and B was due to the high gossypol level in these diets. Shell thickness was found not to be significantly different for the different dietary treatments (Table 7). Eggs produced by birds fed all three diets has approximately the same average yolk score, although the control diet was slightly higher (Table 8). Although diet B contained no yellow corn, it had an acceptable yolk score because of the five percent alfalfa meal in this ration. Eggs produced by pullets fed diets A and 8 had equal, if not better, albumin quality than those fed the control ration as judged by Haugh unit scores (Table 10). It is known that birds fed rations containing cottonseed meal will produce discolored yolk eggs if these eggs are held in cold storage for several months. However, practically all of the eggs were discolored when tested. (The total normal eggs counted, about 5.9% and 1.3% of all eggs tested during January and February, reSpectively.) Of course, some of these eggs were one-day old. 25 Some of these eggs were double-yolked. It was observed that the discoloration was always the same in each yolk. Eggs produced by pullets on diet A (15% cottonseed meal) actually had less discolored yolks than those produced by pullets on diet 8 (12% cottonseed meal). But these differences are not significant. Although the free gossypol level was not determined in these rations, it may be stated that the free gossypol level in these rations was high enough (more than .25% free gossypol) to cause that kind of early discoloration. The cottonseed meal used in this experiment from Buckeye, Cincinnati, Ohio was labeled as shown in Appendix Table 3. Hatchability - The hatchablllty of eggs from pullets fed the control ration was higher than those from pullets fed diets.A and 8 during the entire experimental period (Table 11a). The F ration (Table 11b) indicates that these differences were highly significant (F {L01). Cottonseed products have been reported to cause decreased hatchability (Morgan, 1931; Morgan and Ringrose, 1939; Heywang gt‘gl., 1999; Heywang £3 31., 1950; Naber and Morgan, 1957) but Nelson (1958) reported some improvement in hatchability when barley was treated. This would indicate that the cause of the drop in hatchablllty was the particular sample of cottonseed oil meal used. The F ratio in Table 12b indicates that observed differences in fertility were not significant. The F ratio (Table 13b) indicates that the embryonic mortality (Table 13a) for the three groups was not significantly different. Naber gtflgl. (1957) showed that there were peaks of mortality at the fourth and the eighteenth day of development. Since in this 26 experiment eggs were candied on the 11th day of incubation, no comparisons can be made relative to these results. As shown in Table 1ha the percent of non-hatched chicks from eggs produced by pullets fed diets A and B was higher than those of birds fed the control ration. Differences were found to be highly significant and there was no interaction effect (Table lhb). Fertility - The fertility of eggs from pullets fed diets A and B was as high as those from birds fed the control ration (Table 12a). The F ratio in Table 12b indicates that observed differences in fertility were not significant. Adult Mortality - The F ratio indicates that differences between the number of dead birds in the three groups were insignificant at both levels of probability (Tables 15a and 15b). Average Body Weight -.As shown in Table 16, the gain in body weight of birds fed the control ration was twice as much as that of birds fed diet 8; and was three times as much as that of birds fed diet A. 27 Table 1. Composition of Diet A and Diet B Used in Experiment I Percent of Ration Ingredients Diet 3 Diet Ii Ground barley, 12% protein 28.00 55.00 Corn, ground yellow 25.00 -- Cottonseed meal, solvent, 41% protein 15.00 12.00 Wheat bran, coarse 5.00 5.00 Alfalfa meal, 17% protein 5.00 5.00 Meat scraps, 55% protein 3.50 3.50 Fish meal, 60% protein 2.75 3.00 Animal fats 6.00 7.00 Skimmilk, dried 2.00 2.00 Steamed bone meal 1.00 1.00 Salt, iodized 0.50 0.50 Vitamin-trace mineral mix,* Nopcosol M-4 0.25 0.25 Limestone 6.00 5.75 Total 100.00 100.00 * See the Appendix for the composition of M-4. 28 Table 2. Composition of Basal Ration Used as a Control for Experiment I Ingredient Percent of Ration Corn, ground yellow 59.90 Soybean meal, 45% protein 21.40 Meat and bone meal, 50% protein 2.00 Fish meal and solubles, 60% protein 2.00 Alfalfa leaf meal, 20% protein 2.50 Limestone, ground 6.00 Dicalcium phosphate 1.20 Salt, iodized .25 Fat, animal 4.50 Nopcosol M-4 .25 29 Table 3. Calculated Nutrient Content for the Rations in Experiment I Diet A Diet B Control Protein % 17.71 17.72 17.67 Fat % 8.95 9.23 7.27 Fiber % 6.03 6.76 3.42 Calcium % 3.10 3.08 3.00 Phosphorus % 0.62 0.76 0.68 Arginine % 1.12 1.22 1.02 Methionine % 0.34 0.33 0.35 Cystine % 0.32 0.31 0.25 Lysine % 0.58 0.79 0.90 Tryptophan % 0.17 0.22 0.19 Vitamins (Amt. /1b.) A I.U. 5375 5000 4021 D3 (added) I.C.U. 750 750 750 E mg 3. 6 3. 6 5. 04 B12 mcg 5.00 5.00 6.26 Riboflavin mg 1.56 1.54 2.34 Niacin mg 18.2 19.79 19.89 Pantothenic acid mg 4.2 4.7 5.79 Choline mg 524.00 528.00 537.00 Folic acid mg ? ? .23 Prod. energy Cal/1b 869 811 983 30 Table 4a. Egg Production of Pullets Fed the Three Rations; Calculated as Eggs Per Hen Per Day Diet Replicate Dec. Jgn. Feb. A l .32 .21 .16 2 .27 .20 .13 3 .24 .14 .09 4 .26 .20 08 Average .27 .19 .12 B 1 32 .31 23 2 .37 .32 .24 3 .35 .38 .31 4 .35 .30 .27 Average .35 .33 .26 Control 1 .78 .76 .68 2 .79 .79 79 3 .55 .63 58 4 57 .57 .58 Average .67 .69 .67 31 Table 4b. Statistical Analysis of Differences in Egg Production* Source Sum Degrees of of of Mean F ygriation sgugre freedom square rgtio Row means 42716.3 2 21358.2 7.76 Column means 868194.0 2 434597 157.89 Interaction 9818.1 4 2404.5 0.87 Subtotal 921528.4 8 Within groups 74315.2 27 2752.4 Total 995843.6 35 * Calculated as absolute figures; not as percentages. 32 Table 5a. Feed Consumption for the Pullets Fed the Three Rations Pounds feed per bird per day Diet Replicate Nov. Dec. Jan. Feb. A 1 0. l9 0. 17 0. 18 0. l9 . 2 0. 19 0. 19 0. 19 0. 15 3 0.19 0.17 0.17 0.17 4 0. 19 0. 18 0. 19 0. 19 Average 0. 19 0. 18 0. 18 0. 18 B 1 O. 21 0. 19 0. 20 0. 19 2 0. 19 0. 17 0. 21 0. 18 3 0. 18 0. 17 0. 20 0. 17 4 O. 20 0. 20 0. 20 0. 19 Average 0. l9 0. 18 0. 20 0. 18 Control 1 0. 21 0. 24 0. 26 0. 25 2 0. 23 0. 26 0. 25 0. 27 3 0. 20 0. 21 0. 27 0. 22 4 0. 21 0. 22 0. 27 0. 25 Average -. 21 0. 23 0. 26 0. 25 33 Table 5b. Statistical Analysis of Differences in Feed Intake* Source Sum Degrees of of of Mean F varigtion squares freedom square ratio Row means 1655.8 3 331.2 2.28 Column means 16279 2 8139.5 55.94 Interaction 2417.5 6 219.8 1.51 Subtotal 20352.3 2 Within groups 5236.7 36 145.46 Total 25589 47 * Calculated as absolute figures; not as averages. 34 Table 5c. Feed Conversion Data* for the Three Rations Pounds feed per dozen eggs Diet Replicate Dec. Jan. Feb. A 1 6. l 10. 2 14. 6 2 7.9 10.3 14.9 3 8.5 13.8 22.1 4 8.2 11.3 48.8 Average 7.6 11.2 18.8 B 1 6.9 7.5 10.0 2 5.5 7.7 9.3 3 5. 6 6. 2 6. 9 4 6. 7 7. 7 8. 8 Average 6.1 7.2 8.6 Control 1 3.5 4.0 4.2 2 3. 9 3. 6 4. 1 3 4. 4 4. 9 4. 7 4 4.4 5.4 5.3 Average 3.99 4.4 4.6 * Pen basis. 35 Table 6. Weights of a Random Sample of Eggs Produced by Pullets Fed the Three Rations Average weight (gms)* Diet Replicate Dec. Jan. Feb. A 1 41.2 _-1_- 7.22 40.2 i 6.02 44.9 _-1_- 3.21 2 42.0 i- 6. 12 42.4 i 5.01 43.2 i- 4. 15 3 41.91-4.28 40.01- 3.26 44.61- 2.98 4 38.0;l_-5.04 42.3:4.11 43.6: 7.88 Average 40.8 41.2 44.1 B 1 42.818.21 45.91- 2.25 49.71-4.14 2 41.213.12 39.8: 6.12 47.51-5. 18 3 39.2: 7.14 42.01-4.12 43.31-4.02 4 36.5 i- 2. 26 41.8 i- 3. 08 48.5 _-_1- 6. 13 Average 39.9 42.4 47.3 Control 1 56. 2 _-1_- 4. 20 56. 7 i- 2.06 56. O _-l_- 3. 16 2 54.2 1- 3.39 52.9 _-_1- 3.21 55.5 i- 4.26 3 52.8_-_l-2.28 55.1-_i-4.22 54.91-5.28 4 55.917.02 57.015.19 56.21-6.11 Average 54.8 55.4 55.7 * Eggs from the random sample were weighed individually. 36 Table 7. Shell Thickness of Eggs Produced During January by Pullets Fed the Three Rations Diet No. Replicates A B Control at) ()1) ()1) 1 15.7 15.6 14.6 2 15.9 15.7 16.3 3 14.5 14.1 15.1 4 16. 2 14. 9 16. 2 Average 15.6 15.1 15.6 37 Table 8. Yolk Scores of Eggs Produced by Pullets Fed the Three Rations Diet Repliggte January February A 1 17. 9 18. 1 2 16.9 15.9 3 17. 2 18. 0 4 17.4 16.4 Average 17.4 17.1 B 1 15.9 16.2 2 16.4 15.8 3 16. 2 17. 1 4 17.1 16.4 Average 16.4 16.4 Control 1 17.8 18.4 2 19.8 17.2 3 19. 2 18. 2 4 19.4 19.7 Average 19.1 18.4 38 H 0 9H mH wH HN o H am He Hmuoa i l o N o w o 0 NH oH q H o m o N N o o m m m l i 0 OH N m o o w wH N l l H H w m o o m a H m o o m m 0N mN H N om Hm HmHOH N o o o m m o N m a q l l o H m w o o m m m H o o N w m H o m m N H o m o H N o H 0H m H < .nwm .cmh .nom .cmh .nom .cmh .nmm .amh .HoomHn Hmshoz HoomHv .HoomHe mama emumou .oa uoHQ who: moEum choEEm uszH xumn HmEhoz mwmo oumoHHmoM ammo Hmshoz mxHomiomuoHoomHm Hmuoa mGOHumm wchHmucoolooomaouuou 03H o5“ mom muoHHsm ma woodwoum wwwm smoum Hmeoz can wouoHoomHm mo Hmcaoz .m oHan Table 10. Haugh Unit Scores for the Fresh Eggs Produced by Pullets 39 Fed the Three Rations Diet Replicgte Control Jan. Feb. Jan. Feb. Jan. Feb. 1 85. 8 79. 7 83. 1 84. 1 85. 1 77. 9 2 82.7 77.3 83.9 82.8 82.0 83.2 3 62. 4 82. 7 80. 3 87. 1 68. 9 73. 4 4 73.0 88.2 83.8 79.7 60.8 80.3 Average 76.0 82.0 82.8 83.4 74.2 78.7 40 Table 11a. Hatchability* of Eggs Produced by Pullets Fed the Three Diets December Januaryg Februgry No. of Percent No. of Percent No. of Percent Diet Replicgte eggs hgtch eggs hgggh eggs hggch A 1 38 70.4 17 93.8 16 93.8 2 34 51.6 19 77.8 7 83.3 3 25 25.0 8 85.7 3 66.6 4 38 78.4 24 75.0 3 0.0 Total and av. 135 60.0 68 87.7 29 84.6 B 1 31 72.0 28 89.3 13 92.3 2 36 52.0 35 72.7 16 93.8 3 31 73.9 33 86.7 15 80.0 4 32 83.3 25 92.0 9 50.0 Total and av. 130 70.1 121 84.5 53 82.7 Control 1 48 78.6 25 88.0 10 88.9 2 72 85.1 62 96.6 40 94.9 3 84 76.0 87 95.2 44 ** 4 76 79.2 55 96.1 26 96.2 Total and av. 280 80.6 229 96.1 120 94.6 * Calculated as percent sound chicks of fertile eggs. ** Fertility in this pen was zero. Table 11b. Statistical Analysis of Differences in Hatchability* Source Sum Degrees of of of Mean F variation square freedom square rgtio ROW'means 1109.4 2 554.7 6.35 Column means 992.0 2 496.0 5.68 Interaction 54.5 4 13.63 0.16 Subtotal 2155.9 8 Within groups 2358.4 27 87.35 Total 4514.3 35 * The percentages of hatched chicks were converted according to the tables given by Snedecor, 1950, pp. 449. 42 Table 12a. Fertility of Eggs Produced by Pullets Fed the Three Rations December Janugry February No. of Percent No. of Percent No. of Percent Diet Replicate eggs fert. eggs fert. eggs fert. A 1 38 71.1 17 94.1 16 100.0 2 34 91.2 19 94.7 7 85.7 3 25 80.0 8 87.5 3 100.0 4 38 97.4 24 100.0 3 33.3 Total 135 85.2 68 95.6 29 89.7 B 1 31 80.6 28 100.0 13 100.0 2 36 69.4 35 94.3 16 100.0 3 31 74.2 33 90.9 15 100.0 4 32 75.0 25 100.0 9 88.9 Total 130 74.6 121 95.9 53 98.1 Control 1 48 87.5 25 88.0 10 90.0 2 72 93.0 62 95.2 40 97.5 3 84 29.8 87 24.1 44 zero* 4 76 94.7 55 92.7 26 100.0 Total (incl. 280 73.6 229 66.8 120 61.7 rep 3) Total (excl. rep 3) 196 92.3 142 92.9 76 97.3 * The rooster was replaced by another one after obtaining this result. Results were adjusted according to Cochran, W. G. and G. M. Cox. 43 Table 12b. Statistical Analysis of Differences in Fertility* Source Sum Degrees of of of Ewan F variation squgre freedom squgre rgtio Between means 2605.6 8 325.7 1.06 Within 8331.7 27 308.58 Total 10937.3 35 * Converted according to Snedecor, 1950, 449. 44 Table 13a. Embryonic Mortality of Fertile Eggs Produced by Pullets Fed the Three Diets December Janggry February No. No. No. Diet Replicate eggs % eggg, % eggs % A 1 38 18.5 17 0.0 16 0.0 2 34 16.1 19 11.1 7 16.7 3 25 20.0 8 0.0 3 0.0 4 38 8.1 24 25.0 3 100.0 Total and av. 135 14.8 68 12.3 29 7.7 B 1 31 12.0 28 7.1 13 0.0 2 36 20.0 35 24.2 16 0.0 3 31 8.7 33 6.7 15 13.3 4 32 0.0 25 4.0 9 37.5 Total and av. 130 10.3 121 11.2 53 9.6 Control 1 48 2.4 25 4.5 10 0.0 2 72 1.5 62 1.7 40 0.0 3 84 4.0 87 4.8 44 -- 4 76 1.4 55 1.9 26 0.0 Total and av. 280 1.9 229 2.6 120 0.0 45 Table 13b. Statistical Analysis* of Differences in Embryonic Mortality Source Sum Degrees of of of Mean F variation square freedom squgre grgtio Between means 1111.6 8 138.95 1.18 Within 3169.4 27 117.39 Total 4281.0 35 * Snedecor, G., 1950. p. 449. 46 Table 14a. Percentages of Non-hatched Chicks Related to the Three Diets* December January, February Diet Replicgte Eggs % Eggs % Eggs % A 1 38 11.1 17 6.3 16 0.3 2 34 32.3 19 11.1 7 0.0 3 25 55.0 8 14.3 3 33.3 4 38 13.5 24 0.0 3 0.0 Total and av. 135 25.2 68 5.9 29 6.9 B 1 31 16.0 28 13.6 13 7.7 2 36 28.0 35 3.1 16 6.3 3 31 17.4 33 6.7 15 6.7 4 32 16.7 25 4.0 9 12.5 Total and av. 130 19.6 121 4.3 53 7.7 Control 1 48 19.1 25 0.0 10 11.1 2 72 13.4 62 1.7 40 5.1 3 84 8.5 87 0.0 44 -- 4 76 19.4 55 1.8 26 3.8 Total and av. 280 17.5 229 1.3 120 5.4 * Malformed chicks were included in these percentages. 47 Table 14b. Statistical Analysis of Differences Between Percentages of Non-hatched Chicks Source Sum Degrees of of of Mean F varigtion squgre freedom squgre ratio Row means 1750.5 2 875.3 12.5 Column means 197.5 2 98.7 1.4 Interaction 208.1 4 52.1 0.75 Subtotal 2156.1 8 269.5 Within groups 1885.5 27 69.8 Total 4041.0 35 * Snedecor, G., 1950. p. 449. 48 Table 15a. Mortality During the First Experiment A B Control No. No. No. Replicgge birds % birds % birds % . 1 1 4 1 4 0 0 2 1 4 2 8 0 0 3 0 0 3 12 3 12 4 2 8 1 4 2 8 Total and av. 4 4 7 7 5 5 49 Table 15b. Statistical Analysis of Mortality* Source Sum Degrees of of of Mean F variation squgges freedom square ratio Category means 1.17 2 0.585 0.46 Within 11.50 9 1.278 Total 12.67 11 * Calculated as absolute figures, not as percentages 50 Table 16. Body Weight Gain During the First Experiment for Pullets Fed the Three Rations Average weight in pounds Diet Replicate Initigl 2nd weight Gain (1b) A 1 2.910.26 3.31-0.24 00.4 2 2.810.25 3.31-0.32 00.5 3 2.810.48 3.210.? 00.4 4 2.81-0.32 3.11-0.41 00.3 Average 2. 8 3. 2 00.4 B 1 3.01-0.39 3.610.32 00.6 2 2.910.21 3.51-0.28 00.6 3 2.91-0.25 2.61-0.21 00.7 4 2.810.28 3.61-0.23 00.8 Average 2. 9 3. 6 00. 7 Control 1 2. 8 1 0. 22 4. 2 1- 0. 48 1. 6 2 2.910.27 4.110.32 1.2 3 2.81-0.32 3.810.24 1.0 4 2.810.42 .010.37 1.2 Average 2. 8 4. 0 l. 2 EXPERIMENT II - EXPERIMENTS WITH ADJUSTED LAYING HEN RATIONS An attempt was made to correct the inadequacies of rations A and 8 used in Experiment I (Tables 17 and 18). It was thought that increasing the level of productive energy and increasing the methionine level up to .35 percent might help correct these difficulties. Also part of the cottonseed meal was replaced by linseed meal and higher levels of ferric chloride was used in attempts to overcome the discoloration caused by cottonseed meal. Results 1. Rate of Lay - As shown in Table 19a, pullets on the control ration maintained a higher level of production than did those on diets C and D. The control birds produced almost twice as many eggs as did pullets on either cottonseed diet. These differences were found to be very significant (Table l9b). No interaction effect was involved. 2. Feed Consumption e As shown in Table 20a, pullets fed the control ration consumed about eight percent more than those on diets C or 0. These differences were found to be very significant (Table 20b). 3. Feed Conversion - Feed conversion was slightly improved over that in the first phase of the experiment. As shown in Table 20c, the control ration had a conversion score averaging about 3.5 percent better than that of diets C or D. 4. Egg Quality Determinations - Pullets on diets C or D produced smaller eggs than those fed the control ration (Table 21). These differences were found to be significant (P '.01). 51 52 Shell thickness was similar for eggs produced on all three rations (Table 22). Although some of these differences were significant at .05 level of significance, it may be concluded that these differences were due to the small number of eggs tested. The yolk score was found to be similar in the three different groups (Table 23). As shown in Table 243, the increase in ferric chloride in the rations helped in obtaining more normal yolk eggs than in the first experiment. Although Card (1952) claimed that 0.5 percent ferric chloride in the ration was satisfactory for obtaining normal fresh eggs, about ten to fifteen percent of these rations was .05 percent and .05 percent for rations D and C, respectively (Table 17). When eggs were stored at 609 F. for one week a great increase in the number of abnormal eggs occurred (Table 24b). That remained true also for eggs stored at 609 F. for two and six weeks as shown in Tables 24c and 24d. The normal eggs counted from 39.9 percent to 69.4 percent for eggs produced by pullets fed ration C when stored for one week at 60p F., where it ranged from 42.5 percent to 62.5 percent for eggs produced by pullets fed ration 0. When eggs were stored for two weeks at 609 F. the normal eggs produced by pullets fed ration C ranged from 40 to 70 percent, where it ranged from 53.3 to 75.0 percent for eggs produced by pullets fed ration 0. When eggs were stored for six weeks at 60° F., the normal eggs counted about 37.5 and 50 percent for the sample of eggs produced by pullets fed ration C during March and April, reSpectively. It was 35.7 and 45 percent for eggs produced by pullets fed ration 0 during March and April, reSpectively. 53 The Haugh unit was found to be almost the same for the three groups (Table 25). Hatchability The hatchability of eggs from pullets fed the control ration was higher than those from pullets fed diets C and D, during the entire experiment 1 period (Table 26a). The F ratio (Table 26b) indicates that these differences were highly significant (P‘(.01). Although the ferric chloride was added to the cottonseed meal-containing rations, the hatcha- bility of eggs produced by pullets fed these rations was significantly lower than those produced by pullets fed the control. It was found that fertility of eggs from pullets fed diets A and B was as high as that obtained on the control ration (Table 26a). The-F ratio in Table 26b indicates that there are differences in fertility with respect to dietary treatments. As shown in Tables 27a and 27b, the embryonic mortality for the three groups was not significantly different at both levels of probability (P<.01). As shown in Table 29a, the percent of non-hatched chicks of eggs produced by pullets fed diets A and B was higher than those of birds fed the control ration. Differences were found to be highly significant and there was no interaction effect (Table 29b). The F ratio indicates that differences between the number of dead birds in the three groups were insignificant at both levels of probability (Tables 30a and 30b). As shown in Table 31, the gain in body weight of birds fed diet C and D was five to six times as much as that of the control. This is due to the fact that pullets on the control diet were heavier at the start of the second experiment and had already reached their mature weight. 54 Table 17. Composition of Diet C and Diet 0 Used in Experiment II After Adjustment Percent of ration Ingredients Diet C Diet 0 Ground barley, 12% protein 29.00 52.00 Corn, ground yellow 22.00 -- Cottonseed meal, solvent, 41% protein 12.00 10.00 Linseed meal, solvent, 34% protein 3.70 3.75 Wheat bran, coarse 5.00 5.00 Alfalfa meal, 17% protein 5.00 5.00 Meat scraps, 55% protein 3.50 3.50 Fish meal, 60% protein 2.75 2.75 Animal fats 7.50 3.95 Skimmllk, dried 2.00 2.00 Steamed bone meal 1.00 1.00 Salt, iodized 0.50 0.50 Vitamin-trace mineral mix M-4 0.25 0.25 Ferric chloride 0.07 0.06 Methionine 0.03 0.04 Limestone M E Total 100.00 100.00 55 Table 18. Calculated Nutrient Content for the Rations Used in Experiment II Diet C Diet 0 Control Protein % 17.58 17.54 17.67 Fat % 10.27 10.29 7.27 Fiber % 5.2 7.3 3.42 Calcium % 3.08 3.08 3.00 PhOSphorus % .67 .68 .68 Arginine % 1.114 1.28 1.02 Methionine % .35 .348 .35 Cystine % .3135 .309 .25 Lysine % .75 .84 .90 T'YPtOPha" % .167 .213 .19 Vitamins: A USP 5360 5000 4021 03 added I.C.U. 750 750 750 E mg 3.6 3.6 5.04 812 added mcg 5.0 5.80 6.26 Riboflavin mg 1.58 1.5 2.37 Niacin mg 18.41 19.7 19.89 Pantothenic acid mg 4.14 4.9 5.79 Choline mg 508.5 545.0 537.00 Folic acid mg 7 7 .23 Productive EnergY cal 890.6 839.7 983.0 Ca1orie:protein ratio 50.0 48.9 55.5 IIIII I, 56 Table 19a. Egg Production of Puiiets Fed the Three Rations; Calculated as Eggs per Hen per Day Diet Replicate March c I .22 .35 2 .16 .24 3 .17 .28 ‘1 4.12 13.2 AV. .17 030 0 1 .39 .43 2 .31 .35 3 .32 .35 4 6 8 Av. .35 753 Control 1 .70 .67 2 .78 .79 3 .48 .99 h AV. fig 57 Table l9b. Statistical Analysis of Differences in Egg Production* Source Sum Degrees of of Mean F variation Square freedom square ratio Row'means 387790.8 2 193895.“ 50.55 Column means 5104.2 1 5104.2 1.33 Interaction 12494.0 2 6247.0 1.63 Sub-total 405389.0 5 Within groups 69039.0 18 3835.5 Total 474428.0 23 *Calculated as absolute figures, not as percentage. 58 Table 208. Feed Consumption for the Pullets Fed the Three Rations Pounds feed per bird per dgy Diet Replicate March April C l 0.22 0.22 2 0.20 0.20 3 0.21 0.20 4 0.2 0.24 Average 0.21 0.22 D l 0.26 0.19 2 0.20 0.22 3 0.21 0.21 4 2,25 0.20 Average 0.23 0.20 Control 1 0.26 0.24 2 0.23 0.27 3 0.25 0.24 4 .Qegé 0 26 Average 0.25 0.25 ............... III! 59 Table 20b. Statistical Analysis of Differences in Feed Intake* Source Sum Degrees of of of Mean F variation square freedom g§guare ratio Row means 3339.6 2 1669.8 9.46 Column means 155.1 1 155.1 0.88 Interaction 540.5 2 270.25 1.53 Sub-total 4035.2 5 807.04 Within groups 312618 ‘18 176.49 Total 7212 23 *Calculated as absolute figures, not as averages. 60 Table 20c. Feed Conversion Data* for the Three Rations Pounds feed per dozen eggs Diet Replicates March April c l 11.13 7.38 2 13.8 9.51 3 13.8 8.16 ‘* A M 392 verage 13.72 .27 D 1 7.37 5.20 2 7.30 7.28 3 7.28 6.79 4 Average .7759]- gig-5 Control 1 4.21 4.06 2 3.22 3.98 3 5.73 5.65 4 Average 1%? 111% *Pen basis 61 Table 21. A Random Sample of Weights of Eggs Produced by Pullets Fed the Three Rations Average weight (grams)* Diet Rep. Io. March Syg. April S.d. C 51.2 6.76 51.3 2.18 51.5 2.55 52.9 3.12 53.5 4.59 50.2 3.19 £212 3.03 ‘2213 2.29 Average 51.4 51.9 D 53.1 4.17 54.2 3.17 50.9 3.98 49.9 5.26 53.5 4.81 52.8 2.01 53,} 5.49 2,33 3.80 Average 52.5 52.4 Control 56.8 4.02 57.9 4.11 61.5 3.46 58.2 3.98 57.3 2.32 54.5 5.22 21,} 1.85 5912 2.18 Average 58.4 57.5 iEach egg from the random sample was weighed individually. 62 Table 22. Shell Thickness of Eggs Produced during April by Pullets Fed the Three Rations Diet No. Replicate C 0 Control 9 u H 1 16.2 17.3 15.9 2 15.9 15.9 15.8 3 16.1 16.4 17.8 a 12.2 12.8. 19.2 Average 16.4 16.8 16.5 ..... ..... 63 Table 23. Yolk Scores of Eggs Produced by Pullets Fed the Three Rations Diet Replicates March April C 1 18.2 16.2 2 17.9 17.9 3 16.8 15.8 ‘* 121.1 .1211 Average 18.00 17.4 D 1 17.9 19.5 2 18.2 17.2 3 18.2 18.2 a 1th 1.6.2 Average 17.9 17.9 Control 1 18.4 19.9 2 19.9 18.8 3 17.9 19.8 t. 1.812 32:1 Average 18.8 19.6 aaaaaaa 64 Efirrmwuefimfimsfi HN w o H H N m o HN m on NH : mN m o o o c N N wN m on HH m mN N. 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Haugh Unit Scores for the Fresh Eggs Produced by Pullets Fed the Three Rations Replicates Diet C 0 Control March April March April March April 1 82.7 78.8 86.2 85.6 81.9 79.8 2 83.2 68.7 84.2 82.5 80.2 87.2 3 31.9 32.9 73.9 79.2 59.3 72.9 4 22,4 82,6 81,2 82,2 86,8 88,2 Average 81.8 79.0 82.8 82.6 79.7 82.0 69 Table 26a. Hatchability* of Eggs Produced by Pullets Fed the Three Diets March April ng, Diet Replicates Nogeggs ‘76 No. eggs % No, gqgg % C 1 14 54.5 35 96.2 23 66.7 2 15 50.0 21 71.4 17 70.6 3 15 91.7 29 40.0 20 68.8 t» .2 29.2 2.2 22.1 22 22.2 Total 5 Av. 52 62.8 110 72.2 83 70.6 D l 39 51.7 55 63.6 36 53.3 2 38 66.7 44 44.4 25 72.7 3 27 78.3 44 88.6 27 72.0 i. 22 22.9 22 21.2 22 22.2 Total 5 Av. 139 65.5 193 79.2 114 74.5 1 89 39.7 91 94.3 38 85.7 2 69 91.4 87 88.2 54 84.0 3 57 100.0 51 96.1 22 92.3 h 22 22.2 .62 22.2 29 22.2 Total 5 Av. 280 92.8 295 93.2 144 86.3 *Calculated as %.sound chicks of fertile eggs. 70 Table 26b. Statistical Analysis of Differences in Hatchability* Source Sum Degrees of of of Mean F variation squares freedom sggare ratio Row means 3005.2 2 1502.6 17.29 Col. means 41.2 2 20.6 0.24 Interaction 127.6 4 31.9 0.37 Sub-total 3174.0 8 Within groups 2346.0 27 86.89 Total 5520.0 35 *Converted according to Snedecor, 1950, pp. 949. Table 27a. 71 Fertility of Eggs Produced by Pullets Fed the Three Rations March No. % No. 74 No. T Diet Replicates eggs Fert eggs Fert eggs Fert C l 14 73.0 35 74.3 23 73.3 2 15 80.0 21 100.0 17 88.2 3 15 80.0 29 52.6 20 80.0 a .2 .1222 22 .222 .22 2.2.2 Total 52 82.7 110 71.8 83 81.9 D l 39 74.4 55 67.3 36 75.0 2 38 86.8 44 40.9 25 '88.0 3 27 85.2 44 100.0 27 92.6 h 22 .2122 22 .222 .22 22.2 Total 139 79.1 193 67.4 114 85.9 Control 1 89 97.8 91 95.6 38 92.1 2 69 34.1 37 97.7 54 92.5 3 57 32.5 51 100.0 22 59.1 u 22 222 22 222 32 22.1 Total 280 89.3 295 94.2 144 86.1 ............... ............... ............. 72 Table 27b. Statistical Analysis of Differences in Fertility* Source Sum Degrees of of of Mean F variation sqgares freedom sguare ratio Between 949.6 8 118.7 0.86 Within 3712.0 27 137.48 Total 4661.6 35 *Snedecor, 1950, pp. 449. 73 Table 28a. Embryonic Mortality of £995 Produced by Puiiets Fed the Three Diets March April ng Diet Replicates No. eggs % No. eggs ‘% No. eggs % C 1 14 9.1 35 0 23 11.1 2 15 0 21 4.8 17 0 3 15 0 29 5.3 20 0 '* .3. _°_ ...22 11.2 3.2 .9... Total 5 Av. 52 2.3 110 3.8 83 2.9 0 1 39 0 55 o 36 7.4 2 38 0 44 0 25 0 3 27 0 44 0 27 0 it 22 .2 .22 1.2.2 .22 2. Total 8 Av. 139 0 193 3.1 114 2.0 Control 1 89 0 91 1.1 38 0 2 69 0 87 0 45 0 3 57 0 51 1.9 22 0 1. .62 2 .22 o .22 2 Total 8 Av. 280 0 295 0.7 144 0 74 Table 28b. Statistical Analysis of Differences in Embryonic Mortality*~ Source Sum Degrees of of of Mean F variation squares freedom sqgare ratio Between means 314.5 8 39.31 0.88 Within groups 1208.5 27 44.76 Total 1523.0 35 *Snedecor, 1950, pp. 449. 75 Table 29a. Percentages Non-hatched Chicks Related to the Three Diets March April May Diet‘ Replicates No. eggs % No. eggs % Ng,peggs % C 1 14 36.4 35 3.8 23 22.2 2 15 50.0 21 23.8 17 20.0 3 15 3.3 29 50.0 20 31.3 4 .8 2.2 .22 26.2 22 22.6. Total 5 Av. 52 34.9 110 24.1 83 26.5 D 1 39 48.3 55 5.4 36 14.8 2 33 33.3 44 55.6 25 27.3 3 27 , 21.7 44 11.4 27 28.0 a .22 22.0 50 22.2 .22 22.2 Total 5 Av. 139 34.5 193 17.7 114 23.5 Control 1 89 10.3 91 4.6 38 14.3 2 69 8.6 87 11.8 54 16.0 3 57 0.0 51 1.9 22 7.7 6 .62 2.2 .62 .2._6 .22 22.2 Total 8 Av. 280 7.2 295 6.1 144 13.7 76 Table 29b. Statistical Analysis of Differences between the Percentages of Non-hatched Chicks* Source Sum Degrees of of of Mean F variation squares freedom sqgare ratio Row means 2093.4 2 1046.7 11.71 Column means 141.6 2 70.8 0.79 Interaction 248.4 4 62.1 0.69 Sub-total 2483.4 8 Within groups 2412.5 27 39.35 Total 4895.9 35 *Snedecor, 1950, pp. 449. Table 30a. 77 Mortality during the Second Experiment Diet C Diet 0 Control No. No. No. Repl icates birds % birds % birds % 1 0 0 l 4 2 8 2 l 4 l 4 0 0 3 0 0 l 4 0 0 ‘1 1 .‘t 2 .9 .1. it Total 2 2 3 3 3 3 78 Table 30b. Statistical Analysis of Mortality* Source Sum Degrees of of of Mean F variation sqgare freedom square ratio Category means 0.17 2 0.085 Within 4.50 .12 0.500 0.17 Total 4.67 11 *Calculated as absolute figures; not as percentages. 79 Table 31. Body Weight Gain During the Second Experiment for Pullets Fed the Three Diets Pen Average Weight in Pounds Diet no, 2nd wt. S.d. Last wt. S.d. Gain c 1 3.3 :“0.24 3.9 10.45 +.6 2 3.3 *0.32 4.1 20.48 +.8 3 3.2 to. 28 3.5 £0.32 +.3 4 3,1 350.41 1,; £0.38 1,}; Average 3.2 ' 3.8 +.6 0 1 3.6 10.32 4.2 20.52 +.6 2 3.5 $0.28 4.0 20.52 +.5 3 3.6 10.21 3.9 £0.48 +.3 4 ' Lg £0.23 1,1 20.51 1,; Average 3.6 4.1 +.5 Control 1 4.2 10.48 4.3 20.51 +.1 2 4.1 £0.32 4. 2 20.49 +.1 3 3.8 $0.24 3.9 10.38 +.1 4 5,2 10.37 3.9 £0.48 _._,_1_ Average 4.0 4.1 +.l 80 SUMMARY The hypotheses tested was whether easily available and economical Egyptian-type feed ingredients would perform as well as the American- type ingredients in poultry laying rations. A high level of barley and cottonseed meal feature the former rations. Results obtained from these experiments showed that neither Egyptian rations used performed as well F. as the American one. ' The American-type ration was superior to the Egyptian-type ration in rate of lay, feed consumption, feed conversion, egg size, and ‘I‘tfi¥-, . _' ..,t'.’ hatchability. The Egyptian-type rations were similar to the American- type ration in yolk score, shell thickness, the Haugh unit and embryonic mortality. ‘ Although ferric chloride was added to the rations containing cottonseed meal, a high percentage of the yolks were discolored in the fresh eggs. It is believed that adverse results were due to a very high gossypol level in the cottonseed meal, although the meal was not tested for this constituent. It is obvious that these particular Egyptian-type laying rations cannot be used satisfactorily for market eggs. Further studies are needed in order to overcome those problems caused by the high gossypol levels in laying hen rations containing cottonseed meal. 80 8l Appendix Table l. Prices of Yellow Corn and Barley in Michigan and the United Arab Republic Ingredient Yellow corn Barley Adjusted Barley* Differences Michigan 39.20 37.00 52.86 - 13.66 *Fraps (l9h6) 82 Appendix Table 2. Compositions of Vitamin-Trace Mineral Mix* Per 5 lb, Vitamin A, USP Units ,000,000 Vitamin 03, IC Units ' 1,500,000 Vitamin E, I Units 7,500 Riboflavin, gm 3 El d-Pantothenic acid, gn 5 g} Niacin, gm 20 i Choline chloride, gm 200 Menadione sodium bisulfite, gn -- E; Vitamin 812, gm l0 Zinc bacltracin, gm -- Butylated hydroxy toluene, gm ll3.h Manganese, % 2.4 Zinc, % l.l Iron, % 0.8 Copper, % 0.08 Iodine, % 0.0h8 Cobalt, % 0.008 *Nopcosol M45, Nopco Chemical Company, Harrison, New Jersey 82a Appendix Table 3. Labeling of Cottonseed Meal Used in the Experiment hi Percent Protein Solvent Extracted Cottonseed Meal with Added Hydrolyzed Cottonseed Oil Crude protein Min. hl% Crude fat Min. 2% Crude fiber Max. l#% Nitrogen free ext. Min. 27% LITERATURE CITED Ahmed, I. A. and M. F. Badr, 1953. A study of animal feeding methods in Egypt and treatments of defects therein. J. Agr. Sci. (Cairo Univ., in Arabic) Almquist, H. J. and F. U. Lorenz, 1933. "Pink whites" in stored eggs. U. 5. 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