OVERDUE FINES ARE 25¢ PER DAY PER ITEM Return to book drop to remove this checkout from your record. EFFECT OF FEATHER MEAL AS A SOURCE OF PROTEIN ON THE PRODUCTION OF LAYING HENS By Abolghasem Golian 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 1979 ABSTRACT EFFECT OF FEATHER MEAL AS A SOURCE OF PROTEIN ON THE PRODUCTION OF LAYING HENS By Abolghasem Golian It has been proposed that more hydrolyzed feather meal can be used in the diet of laying hens than in that of broilers. It was therefore of interest to evaluate the effect of different levels of hydrolyzed feather meal on feed utilization, egg production, feed-efficiency, weight gain and egg quality as measured by Haugh units. Replicate pens of Single Comb White Leghorn (SCNL) hens were fed rations containing 3, 6 or 9 percent hydrolyzed feather meal with and without supplementation of amino acid (lysine and methionine). The birds were 20 weeks of age when placed on treatment and data were collected for 20 weeks. The birds were confined in individual cages and exposed to 14 hours of light:lO hours of dark throughout the experiment. The experiment was designed so that the orthogonal test for determining linear or quadratic relationship between the hydrolyzed feather meal and interest factors could be utilized. There was a significant difference (P< 0.05) between birds which received the 3 and 9 percent hydrolyzed feather meal with no supplementation of amino acid in egg out-put and egg weight with a Abolghasem Golian linear decrease from 3 to 9 percent. There was no significant difference in egg production and egg weight (P:>0.05) between the control group and those fed 3, 6 or 9 percent hydrolyzed feather meal with supplementation of methionine and lysine. There was no significant difference (P>'0.05) between the control and treated groups in feed- intake. feed efficiency, Haugh units or mortality. All the observed factors except mortality changed significantly (P< 0.05) by periods in various treatments. Mortality occurred at random in all treatments. ACKNOWLEDGMENTS The author wishes to express his appreciation to Dr. C. J. Flegal and Dr. T. H. Coleman as his academic and research advisors, for their guidance and interest in this study and their helpful suggestions in the preparation of this manuscript. Special thanks is due to Dr. H. C. Zindel as Chairman of the Poultry Science Department who made available the funds and facilities that I needed. Most sincere appreciation is extended to Dr. C. R. Anderson for his help in computer programming and Dr. J. L. Gill for his help in statistics. The author also appreciates Mr. A. Ansari as a fellow graduate student who was invaluable in helping him during the experimental procedure. ’ Finally. the author is indebted to the College of Agriculture, Ferdowsi University, Mashhad, Iran for providing him the scholarship to work toward his master of science degree in the Department of Poultry Science at Michigan State University. ii TABLE OF CONTENTS LIST OF TABLES .......................... LIST OF FIGURES ......................... INTRODUCTION ....................... REVIEW OF LITERATURE ................... Broilers ........................ Feather Meal ...................... Agricultural and Industrial Uses ............ Soil Fertilizer .................. Feed ........................ Factors Affecting Utilization of Low Quality Feather Protein by Poultry .............. Feed Intake .................... Digestibility ................... Chemical Structure of Feather Protein ......... Chemical Studies on Keratin (Wool and Feather) ..... Processing Effects on Low Quality Feather Protein . . . Cooking ...................... Drying ....................... Fat Extraction ................... Grinder ...................... OBJECTIVES ........................ EXPERIMENTAL PROCEDURE ..... . ............ Experimental Design ..... . ............ Treatments ...................... Management and Feeding Program ............. Statistical Analysis .................. iii Page -—I 100000000 0301050000 w Chapter Page V. RESULTS AND DISCUSSION .................. 25 Production ....................... 25 Egg Height ....................... 28 Feed Consumption .................... 32 Feed Conversion .................... 37 Haugh Units ...................... 42 Mortality ....................... 42 Weight Gain ...................... 50 General Discussion ................... 50 The Effect of Feather Meal as a Source of Protein on the Production of Laying Hens ........... 52 Summary ...................... 52 APPENDIX ............................. 54 LITERATURE CITED ......................... 61 iv Table 01-500“) 10. ll. l2. l3. T4. 15. LIST OF TABLES Chicken Meat, Broilers: Production in Top Ten Companies ........................ The Top Eight Broiler Firms in the United States . . . . Composition of Feather Meal ............... Composition of Control Diet Used for the Experiment . . . . Composition of Rations With Different Levels of Hydrolyzed Feather Meal ................. Calculated Rations Nutrient Composition ......... The Effect of Dietary Supplementation With Methionine, Lysine and Mixture of Essential Amino Acids to HFM Diets on Performance of Laying Hens ........... The Percentage of Production in the Control Group, Supplemented and Unsupplemented Groups ......... The Average of Egg Weight in the Control Group, Supplemented and Unsupplemented Groups ......... The Average of Feed Consumption in the Control Group, Supplemented and Unsupplemented Groups ......... Nutrients Consumption .................. The Average Feed Efficiency in the Control Group, Supplemented and Unsupplemented Groups ......... The Average Haugh Units in the Control Group, Supplemented and Unsupplemented Groups ......... Analysis of Variance by the Use of Split-Plot Repeat Measurement for Egg Production .......... Analysis of Variance by the Use of Split-Plot Repeat Measurement for Egg Weight . . .......... Page 21 22 23 26 3O 35 36 38 44 47 54 55 Table Page l6. Analysis of Variance by the Use of Split-Plot Repeat Measurement for Feed Consumption ......... 56 17. Analysis of Variance by the Use of Split-Plot Repeat Measurement for Feed Conversion ......... 57 18. Analysis of Variance by the Use of Split-Plot Repeat Measurement for Haugh Units ........... 58 19. Analysis of Variance by the Use of Split-Plot Repeat Measurement for Mortality ............ 59 20. Analysis of Variance by the Use of Split-Plot Repeat Measurement for Average Weight Gain ....... 60 vi LIST OF FIGURES Figure Page 1. Broiler Feather Processing ............... ll 2. Percent Production Response by Period for Control and Supplemented Groups ................ 27 3. Percent of Production Response by Period for Control and Unsupplemented Groups ............... 29 4 Percent of Production Response to HFM ......... 3l 5 Egg Weight_Response to HFM in Diets .......... 33 6 Egg Weight Response to Periods ............. 34 7. Feed Consumption Response to HFM in Diets ....... 39 8. Feedélntake Response to Period ............. 40 9 Feed Efficiency Response by HFM ............ 41 lO. Feed Efficiency Response by Period ........... 43 ll. Haugh Units Response by HFM .............. 45 l2. Haugh Units Response by Period ............. 46 l3. Effect of Treatment on Mortality . ........... 48 l4. Mortality Response by Period .............. 49 15. Weight Gain Response by Treatments ........... 51 vii CHAPTER I INTRODUCTION Unavoidable problems of by-product and waste disposal have plagued the poultry slaughtering industry for many years. The problems have become more acute with the development of large scale commercial plants for poultry slaughter. At least 7 kg of feathers are left for every 100 kg of live birds (Mitchell, 1926, 1931). Furthermore, feathers are made up of 80 to 85 percent protein. Protein supplements in the poultry diets represent one of the major items of cost. Alternate sources of these supplements might have a beneficial effect on production cost providing they are available at a competitive price and are acceptable to poultry. Feather protein was found to be deficient in tryptophan, methionine, histidine, and lysine (Routh, 1942). It is known that by adding the essential amino acids missing in feather meal one can increase the percentage of this ingredient in the diet without decreasing maximal production (Moran et al.. 1969; Bhargava and O'Neil. 1975a; Daghir, 1975). But its usage in feed still is limited because the feather meal is poorly absorbed (Summers et al.. 1965), and it also affects the palatability of the feed and decreases feed intake (Van and Payne. 1977). Both growing and laying birds may be expected to perform below normal as a result of use of feather meal. It is also possible that these protein sources may be put to more extensive use with the hen than with the chicks or broilers. This expectation is based on the knowledge that the protein consumed for maintenance of the hen is higher than for the broiler. The laying hen, because of its larger absolute body size and normally lower rate of net nitrogen accumulation (body tissue plus egg formation) will have a greater amount of its feed devoted to maintenance than will the rapidly growing broiler. This is due to the fact that maintenance in the hen is primarily concerned with synthesis of feather keratin (Leveille and Fisher, 1960) and keratin feed meals have an amino acid pattern nearly perfect to meet this need. It appears that feather meal might be used to a greater extent with the laying bird. CHAPTER II REVIEW OF LITERATURE Broilers The broiler industry has expanded tremendously over the past 20 years throughout the world. In 1978, Holly Farms Company broke the billion pounds a year barrier on the live weight basis (Broiler Industry, December 1978). The top ten countries which lead in broiler production produce more than 9 billion kilograms of meat annually and 700 million kilograms of feathers as indicated in Table 1. Table 2 shows the weekly slaughter of the top eight broiler companies in the United States. Feather Meal Mitchell (1926. 1931) reported the feather yield of Leghorns and Rhode Island Reds ranged from 6.6 to 7.6 percent with an average of 7.0 percent of live weight. About 23 percent of the live bird is blood and offal. Feathers are composed of quills. barbs, barbules. and the barbicels, which in the broiler feather processing are cooked and ground together to produce feather meal (Humbert, 1957). The American Association Feed Control Office (AAFCO) defines hydrolyzed poultry feather as the product resulting from the treatment under pressure of clean undecomposed feathers from slaughtered poultry Chicken Meat, Broilers: Table 1 Production in Top Ten Countries (in thousand of metric tons) 1975 1976 1977 1978 United States 3,666, 4,109 4,272 4,604 Japan 616 701 785 825 Brazil 349 552 632 676 Spain 561 617 652 688 France 517 536 561 596 United Kingdom 444 550 555 560 Italy 495 496 500 515 Soviet Union- 190 190 387 500 Canada 291 329 340 355 Netherlands 267 292 300 292 Total 7,396 8,372 8,984 9,611 Reports of U.S. Agricultural Attaches, August 1978, Foreign Agriculture Circular, USDA. Table 2 The Top Eight Broiler Firms in the United States Weekly Slaughter Firm Plant Locations (Head) Gold Kist Boaz, Trussville, Ala.; Oak Live, 5.0 million F1a.; Athens, Carrollton, Elligjay, or more Holly Farms Tyson Foods Lane Poultry Company Perdue Foods Valmac Industries Central Soya Company Conagra Ga.; Durham, N,C.; Jasper, Texas; Fayetteville, Ark. Wilkesboro, Monroe, Hiddenite, N.C.; 4.7 million Center, Seguin, Texas; Glen Allen, or more New Market, Temperanceville, Va.; *5.2 marketed (*Includes broilers packaged by Holly but processed by others) Berryville, Green Forest, Nashville, 4.0 million Rogers, Springdale, Ark.; Lola, Kan.; or more Monett, Mo.; Shelbyville, Tenn.; Springdale, Ark.; Cumming, 6a.; Dobson, Robbins, N.C. Ashland, Blountsville, Gadsden, 3.3 million Heflin, Ala.; Grannis, Ark.; or more Broken Bow, 0k1a.; Fort Worth, Mt. Pleasant (1/2), Wacco, Texas; Dexter, Mo. Felton, Georgetown, De1.; Salisbury, 2.7 million Md.; Lewiston, N.C.; Accomac, Va. or more Clarksville, Dardanelle, Pine Bluff, 2.6 million Waldron, Ark.; Logansport, La.; or more Muskogee, 0k1a.; Carthage, Nacogdoches, Texas; SwiftzBloomer, Ark. (May 1) Athens, Canton, Ga.; Monroe, 2.5 million Robersonville, N.C.; or more Chattanooga, Tenn. Athens, Enterprise, A1a.; Dalton, 6a.; Arcadia, La. Broiler Indu stry, December 1978, p. 17; by permission. free of additives and/or accelerators. Not less than 75 percent of its crude protein shall consist of digestible protein by the pepsin digestibility method (Proposed, 1961; adopted by AAFCO, 1965). Agricultural and Industrial Uses Soil Fertilizer In the past untreated offal and feathers have been applied to soil as a fertilizer; this method of disposal was generally unsatisfactory (Naber and Morgan, 1956). feed. The Keratin proteins have been considered to be of little or no nutritional value because of poor digestibility (Mangold et al.. 1930); however, the feather is very high in protein and relatively rich in energy, mineral and fat (Table 3). The development of a method by Binkley and Vasak (1950), for processing feathers into friable, high density meal opened the way for feather meal to successfully supply part of the dietary protein in poultry (Moran et al.. 1966, 1967a, 1967b, 1968; Daghir, 1975). In rations for ruminants, feather meal protein is equal to any protein supplement on a "per unit of protein" basis because amino acid balance doesn't have the same importance as it has in non-ruminant rations. It can be used to replace all of the plant protein supplement when the animals are given a chance to become accustomed to it (Kennett et al.. 1972; Morrison, 1971). Table 3 Composition of Feather Meal NRC Scott b Nutrient Name Units Analysisa Analysis Metabolizable energy cal/kg 2,360 2,310 Protein % 86.4 85 Arginine % 5.42 5.6 Glycine % 5,31 _- Histidine % 0.34 -- Isoleucine % 3.26 -- Leucine % 6.72 -- Lysine % 1.67 1.5 Methionine % 0.42 0.5 Cystine % 4.00 3.0 Phenylalanine % 3.26 -- Threonine % 3.43 -- Tryptophan % 0.50 0.5 ' Valine % 5.57 -- Total fat % 3.30 2.5 Total fiber . % 1.00 1.5 Calcium % 0.33 0.2 Available phosphorous % 0.55 0.6 Sodium % 0.71 -- Thiamine mg/kg 0.10 -- Niacin mg/kg 27.00 24.0 Riboflavin mg/kg 2.10 2.0 Pantothenic acid mg/kg 10.0 11.0 Vitamin B-12 mg/kg 0.078 -- Choline mg/kg 891.00 900.0 Pyridoxine mg/kg -- -- Folacin mg/kg 0.20 -- Biotin mg/kg 0.44 -- Potassium % 0.44 -- Magnesium % 0.20 -- aNational Research Council, 1977. b Scott, Nesheim and Young, 1976. Factors Affecting Utilization of Low Quality Feather Protein bngoultcy The two most important factors which influence utilization of low quality feather protein by animals are feed intake and digestibility. Feed Intake When protein feed supplements are in short supply feather meal might be a suitable ingredient in feed for laying hens and broilers (Sullivan and Stephenson, 1957; Bhargava et al., 1975b). However, the feed intake will decrease as the level of feather meal is increased in the diet (Bhargava and O'Neil, 1975b; Moran et al., 1969). Van and Payne (1977) pointed out that the higher amino acid supplementation of feeds with protein sources deficient in some amino acids is much more critical on low feed intake. Digestibility Approximately 85 to 90 percent of the protein from feather meal comes from keratin (Harrap and Woods, 1964). The keratins are classified in the sclero-protein group because of their insolubility in aqueous solvents (Fruton and Simmonds, 1960). This keratin must be hydrolyzed in order that it may be digested by animals. According to the AAFCO official definition for hydrolyzed feather meal, 75 percent of its crude protein shall consist of protein that is digestible by pepsin. Chemical Structure of Feather Protein The chemical structure of keratin consists of chains of amino acids joined by peptide bonds formed by a combination of the amino group of one acid to the carboxyl group of the next. A large number of amino acid residues are linked into a single molecule. Assuming all the protein of feather is keratin, the molecular weight is 10,400 (Harrap and Woods, 1964). In their native state, these molecular chains are arranged in an orderly manner, stabilized primarily by hydrogen bonds which can be broken by chemicals or heat. When these bonds are broken, the protein loses its original native properties and is denatured. The great stability of keratin is due to a cystine disulfide cross link, a central bond between the two sulfur atoms (covalent bonds); 8.8 percent of this protein is cystine (Block and Bolling, 1951). The structure of feather keratin may be visualized as extended chains of amino acids bonded together by hydrogen bonds and cross linked by disulfide bonds. Chemical Studies on Keratin (Wool and Feather) The insolubility of keratins and their resistance to digestion by enzymes have to be explained on the basis of protein structure. Many investigators have studied these properties and have attempted to alter them. Kuhne in 1877 observed that the keratin of hair was 10 made digestible by pepsin when surface area was increased by mechanical means. Powdered wool was used by Harris and his coworkers (1932) in studies on isoelectric point, the amino nitrogen content (Kanagy and Harris, 1935). Routh et a1. (1938) observed that after wool was ground the powdered material was digested by both pepsin and trypsin. An appreciable fraction of the nitrogen and sulfur of powdered keratin was soluble in water. Scott and Payne (1926-1928) showed that hydrolyzed feathers improved egg production in one year.but not in the second year, and these researchers concluded that feathers added nothing of importance to poultry diets and the cost of hydrolyzing was too great to allow the use of feather keratin in practical poultry feeds. Balance studies conducted by Mangold and Dubiski (1930) failed to show any digestion of white goose feathers by cats, owls, dogs, and rats. Thus, it appeared that native feather keratin was not only seriously deficient in certain amino acids but also poorly digestible. It was found that powdered feathers were capable of supporting moderate growth in the young rats when supplemented with methionine, lysine, tryptophan and histidine (Routh, 1942). Processing Effects on Low Quality Feather Protein The discovery of a processing method for poultry by-products opened a way for researchers to use more of these products as feed. Figure 1 shows the normal processing. 11 new ~98 .3: a. 38 «.3 my 5305 98¢ .00 OOOOOOOOOOOOO. o\. 8 . 2.8 e 09. 3% V .2 n. t oNpm .225 n. «3.0 IJJE 358. 2 8 m. § 3 A.R A _.\.v_ _ 6m 95 an; a. -d _o\.mm _ 5295 2 m3. IIOOOOOOIOOOOOOOO _ $3. 2.8 a. om. 7.3 _ .2 a. to Tam _ .033 a. 35 «we 652: e 03.. .u.o .coumcwsmm: .flvmm0 on \fim.we n .355" u totem n .8300. u ” “00000.00" “00000.00 OOOOOOOOOO 50¢” ; a. thm bod”; n. whsrpn IBoommmmhmmz 400.000.00.000 Ill]. .0950 _ firm. U..Ow £09m .30.: .2 .83 .o a; .moEb an _ mam 3mm! "am .m . a...» $3 .0300 o .22.. b. m9 .32.. Jul. .2. «on... a. m. 22.2.. a. co. 12 Cooking The quality of by-products can be influenced during cooking by either physical or chemical means. Elevated temperature and prolonged cooking time promote chemical changes, especially oxidation of the fat. It is recommended that the temperature during cooking definitely not be allowed to go over 250° F. Also, for maintaining fat quality, cooking should not be continued for more than two hours. The fact that poultry offal usually takes longer to cook than wastes from other animals makes it more difficult to obtain a high-quality fat from poultry offal. It has been claimed that partially cooking poultry offal in a cooker and completing the drying in a separate dryer will give better fat quality. It also appears that higher temperatures.(>250° F) cause the by-product meal to have a low digestibility and a low nutritive value (0thmor, 1954). Studies on flame drying and steam-tube drying of fish meal, where presumably the flame drying caused higher temperature in the meal, did not show any difference in digestibility of the fish meal (Grau et al., 1955; Almquist, 1956). Overcooking to the point of burning the tankage and forming a rubbery carbonized product with low feed value is pos"\1e if cooking temperatures are not controlled. Overcooking will also reduce the yield of fat during subsequent pressing. Physical changes in quality result from agitation and over- cooking. Some renderers feel that less fines are produced (which subsequently press out with fat) if the agitator speed is kept below 13 20 rpm instead of the commonly used 38 rpm (Humbert, 1957). The lower agitator speed will increase the required cooking time. Overcooking and making the tankage too dry causes more fines than higher agitating speed does. "In summary, one can qualitatively state that low cooking temperatures, low agitation speed, and short cooking time will tend to give a better quality product" (Humbert, 1957). Final moisture content is important in that it indicates whether the cooker batch is overcooked or undercooked. Eight percent moisture in the tankage is usually assumed to be the optimum moisture content. The effect of undercooking results in too much moisture and poor pressing characteristics of the tankage. Drying Separate dryers are used to increase production by reducing the time the tankage must be held in the cookers. Generally, the tankage must be removed from the cookers in half the time normally required if separate drying is used. The use of separate dryers is quite common for feather meal. Use of separate dryer probably reduces the agitation and grinding effect on the tankage (Humbert, 1957). It is possible to overheat the tankage in a dryer. However, the drying temperature had to be lowered to reduce the obnoxious odors given off. (When the material-exit temperature was reduced from 160- 170° F down to 140-150° F, the amount of the odor was reduced and the color of the feather meal became lighter, indicating better quality [Harstad, 1956].) _..‘ 14 Fat Extraction Storage of offal before pressing to extract fat will reduce the quality of the by-product meal and fat. The fat present in the offal tends to be oxidized and become rancid. If a suitable stabilizer (antioxidant) has been added during the cooking, the problem of ran- cidity developing during storage of the tankage before pressing is reduced. A high moisture content of the tankage promotes hydrolysis 1 and thus increases the chances of spoilage. The pressing itself does not alter the quality of fat significantly, unless the equipment is not kept clean. The moisture in the fat is primarily influenced by the cooking operation. If the pressing is not properly operated, the by-product meal can have too high a fat content, which may promote spoilage of the meal. This ‘high fat content will also make the subsequent grinding operation more difficult. Grinder Grinding must be performed at some stage before poultry by-products are compounded into animal feeds. In some cases small renderers may not grind any of their by-products but sell them as pressed cakes or unground feather meal and unground dried blood. Hammermills are used to obtain the finished grind, commonly to 8 or 12 mesh. In some instances, separate crushers are used to break the pressed cake and obtain a material that can be fed to the hammermill. However, many hammermills are manufactured with a built-in cake crushing mechanism. The power required to grind a specific 15 quantity of material will depend on the particle size to which it is ground. A relatively coarse material may be produced with only 10 horsepower per ton per hour of capacity, while a relatively fine material may require 25 horsepower per ton per hour of capacity (Humbert, 1957). Feather Meal in Poultry Rations 0n the basis of the amino acid content of keratins given by Graham et a1. (1949) and from the amino acid contents of the other dietary ingredients (Almquist, 1952), it is apparent that keratin from feathers can substitute for a large portion of soybean oil meal in commercial chick rations without distrurbing the amino acid balance of the ration (Wilder et al., 1953; Sullivan and Stephenson, 1957; Naber et al., 1961; Poppe, 1965; Vogt and Stute, 1975). Since feather meal has a good replacement value for the chicks at high protein dietary levels (20 to 23 percent) and poor substituting ability at low dietary levels (15 percent), Sibbald et al. (1962) suggested that feather meal was being used as a source of non-specific nitrogen. Studies conducted by Naber et a1. (1956) showed that feather meal and poultry meat scrap were capable of replacing 5 percent or one-fourth of the protein in broiler rations containing large amounts of soybean oil meal and corn fortified with fish meal, dried whey product, methionine, minerals, vitamins and antibiotics. McKerns and Rittersporn (1958) substituted keratin for 50 percent of the soybean oil meal equivalent to 25 percent of the 16 total diet, keratin meal improved the feed efficiency. Wilder et a1. (1953, 1955) also found a growth response from feather meal when fed to chicks. Gerry et a1. (1954) and Morris and Balloun (1971, 1973) reported that feather meal substituted for one-fourth of the total protein in the diet had no adverse effect on growth of broilers. In the studies by Burgos et a1. (1974) and Bhargava et al. (1975a), poultry offal meal and hydrolyzed feather meal were blended to evaluate poultry by-product and hydrolyzed feather meal (PBHFM) as a protein supplement in the diet of broiler chicks. There were no adverse effects on body weight and feed efficiency when PBHFM was incorporated into the diet to the level of 10 percent. But the addition of either 15 or 20 percent (one-fourth of the total protein) PBHFM significantly depressed growth and feed efficiency. Very few studies have been reported on the use of feather meal in rations of laying hens. Moran et a1. (1969) studied the effect of feather meal and hog hair meals (keratin) as sources of protein for laying hens with 10 percent protein based on 5 percent from soybean and 5 percent from corn. When the diet was supplemented with methionine the diet was shown capable of supplying the estimated minimal essential amino acid needs of laying hens (Leveille and Fisher, 1960). However, the diet was unable to support maximal performance. Also, by adding 5 percent keratin meals to the basal diet, it was found that several measured parameters of performance were improved comparable to the basal diet, but that supplemental methionine was necessary for maximal production and egg weight. Van and Payne (1977) conducted two 17 experiments. In the first one, the control diet was compared with five diets that contained 7 percent hydrolyzed feather meal (HFM) which varied in lysine content. The basal diet was very deficient in lysine. As lysine supplementation was increased, egg production, feed consumption and live-weight increased, and improved feed effi- ciency occurred. In the second experiment, the control diet was compared with five diets that contained 7 percent HFM which varied in methionine, lysine and tryptophan. When the diet with HFM was supplemented with all three of the above amino acids to a level higher than that in the control diet, a satisfactory level of performance was obtained. Their study in the methionine supple- mentation for increasing egg output was in agreement with the work of Moran et a1. (1969). The effect of tryptophan supplementation was . explained partly by the deleterious effect of HFM in poultry diets as indicated by Daghir (1975) and Macalpine and Payne (1977). Moran also concluded that the lower egg production confirms the suspicion that not all lysine measured by amino acid analysis in HFM is available to poultry. CHAPTER III OBJECTIVES Interest in the use of feather meal for poultry feed is increasing as the prices of quality protein feeds increases. At present, millions of pounds of feather meal available annually are utilized as feed. The potential use of feather meal is worthy of ' consideration in view of the low price of its protein unit content. Feather meals are essentially protein feeds, low in fiber and rela- tively high in calcium, phosphorous and fat. Tsang et a1. (1963) incorporated hydrolyzed feathers in a series of broiler rations. Based on actual chemical analysis, it was calculated that approx- imately 40 kilograms of hydrolyzed poultry feather and 35 kilograms of ground yellow corn would supply the same amount of protein and productive energy as 70 kilograms of dehulled soy-bean meal and 5 kilograms of stabilized animal grease. Feather meals cannot be the sole source of protein in poultry rations, because of the imbalance of amino acids and because the pro- tein of feather meal is poorly absorbed (Summers et al., 1965). The objectives of this study were to determine: 1. the effect of utilization of feather meal on production of laying hens. 2. to what level commercial feather meal can be used without decreasing egg production and/or egg quality. 18 CHAPTER IV EXPERIMENTAL PROCEDURE The experiment was conducted at the Michigan State University Poultry Science Research and Teaching Center, from December 15, 1978 to May 3, 1979. Experimental Design Twenty-week—old Single Comb White Leghorn (SCWL) pullets were sorted into groups of similar weight. Pullets were selected (omitting the extremes of heavy- and light-weight ones), weighed and distributed into groups according to average weight. They were housed in individual laying cages (19 centimeters wide X 36 centimeters high X 33 centimeters deep) with double deck. A chart of random numbers was used so that each part of a deck would have each particular replicate. This was done to reduce the chance of having a treatment group on a particular deck be more favorably treated than those placed elsewhere. Treatments The experiment consisted of seven treatments with three replicates each. There were eight birds within each replicate with a single feeder. The treatments were designed to consist of seven rations as follows: 19 20 1. Control ration (A) with no feather meal1 in it. 2. Ration (B) with 3 percent feather meal supplemented with amino acids. 3. Ration (C) with 6 percent feather meal supplemented with amino acids. 4. Ration (0) with 9 percent feather meal supplemented with amino acids. 5. Ration (E) with 3 percent feather meal without supplementation of amino acids. 6. Ration (F) with 6 percent feather meal without supplementation of amino acids. 7. Ration (G) with 9 percent feather meal without supplementation of amino acids. The control diet used for this experiment is shown in Table 4. It is basically a corn-soybean type diet satisfying all the nutrient requirements of the laying chicken. In all rations total energy and protein content of the diet were adjusted by changing the amount of corn and soybean. Ration G was further adjusted by changing the amount of the fish meal in the diet. Therefore all rations provided isocaloric and isonitrogenous contents (Table 5). The composition and calculated analysis of nutrients of the experimental rations are shown in Table 6. 1Feather meal contained minimum 85 percent protein, minimum 2 percent fat and minimum 6 percent moisture manufactured by Badger By-Products Company, 511 E. Menomonee Street, Milwaukee, Wisconsin 53202. 21 Table 4 Composition of Control Diet Used for the Experiment Ingredients Percentage of Rationa Corn 59.57 Soybean 49% ' 18.50 Fish meal MH 3.00 Wheat middling 5.00 Fat hydrolyzed 1.80 Alfalfa 17% 3.00 Limestone 6.70 Methionine DL 0.006 Dical 1.50 Salt 0.427 Premixb 0.50 aAs fed basis. bSupplies the following per kilogram of ration: 8800 USP vitamin A; 2750 ICU vitamin 03; 7.7 mg ribo- flavin; 13.64 mg pantothenic acid; 27 mg of niacin; 429 mg choline chloride; 1.1 mg folic acid; 0.0013 mg vitamin B-12; 5.5 IU vitamin E; 1.65 mg menadione sodium bisulfite; 64 mg manganese; 1 mg iodine; 4 mg copper; 251 mg cobalt; 50 mg zinc; 25 mg iron; 500 mg magnesium. 22 .muwoe ocean mo copumuewEmpaasm uaoguwz Fame cmguowm vm~apocuxz u 1ze= .mcmom ocean Go cowumueoEmpnazm sum: poms cmsumme um~zpoeuxz u +zu: om.o om.o om.o om.o om.o om.o om.o stmca -1 11 1- m~.o moo.o 11 11 enema. oo.m oo.o oo.m oo.m oo.o oo.m - Fame Locummu ~N¢.o ~N¢.o NN¢.o m~.o mmm.o mmm.o ~N¢.o .upem om.p om.p om.p om._ o¢.F om.~ om.p .muwo moo.o moo.o ooo.o mno.o umo.o _mo.o o .o mcweopgumz on.o o~.m o~.m o~.e o~.m on.o o~.m meoummem. oo.m oo.m oo.m oo.m oo.m oo.m oo.m NAP nepaep< om.F om._ om.p om.o o~.o I o~.p om.~ «Napoeuas an; oo.m oo.m oo.m oo.m co.m oo.m oo.m .uwe pews: oo.m oo.m oo.m oo.m oo.m oo.m oo.m :2 Fame cmwd um.o ~w.¢ mo._P o~.~ o~.o om.- om.wp Nae cmmnzom o¢.o~ m~.~m ¢¢.mo mm.uo me.mm oo.mm mm.mm ccoo -2”... .3 .2... .3 1...... em +2.: .3 1...: .2. +2.... .8 2. 553.. 833.88.. am. copumm Ad. comumm Am. cowumm Ac. cowpmm Au. cowuma Am. :owumm Pocucou comm Asezv Fem: Locumme um~xFoeuxz mo m_m>w. acmcmmwwo new: Ag. meowumm eo eowuwmanou m «Fame 23 .cmxowco we» mo cowuwcpaz “asap .m:=o> use Ewmgmmz .uuoom Ease muesmwun .xgupzoa we mangmcwacmc acmwcuaz ”sump .queaou goeemmmm chopumz use soc» umuoaeuxm mew: macaw?» mmmgpm bk 11 11 oN.o mem.o mN.o p~.o pm.o FN.c FN.o Ezwtom 1- 1- m¢.o m¢.o w¢.o m¢.o w¢.o w¢.o w¢.o ngocnmoca .pmw>< 11 11 ~—.m e~.m mp.m Np.m ~—.m N—.m -.m Eswupmu up.o _—.o mm~.o mm~.o nm~.o mp.o m_.o om.o mm.o suscepaxyh 11 -1 mpm.o mvm.o mmm.o mm.o No.o oo.o mm.o who + gum: em.o um.o mom.o mm.o mum.o Fm.o pm.o pm.o pm.o An 1 gum: mn.o om.o mpw.o oo.o 55.9 mo.o me.o ow.o em.o mamme ms.o oc.o om.mp oe.o— mo.mp mp.mp mm.m~ mm.~— mm.np ammuoga mango an m mm. mm. mm. mm. mm mm. mu 0- P. “U NU ”WU “U "U ”U U0 . _ . + + +1! \II.. M m m m. m m w Aeomuma mo NV =o_uwmoQEou bemwcuzz eomuom umumpzu_mu m mpnmh 24 Management and Feeding Prggram The house was ventilated (0.25 to 4.0 CFM/bird) and incandescent light was supplied 14 hours a day at 0.5 ft. candles intensity. A11 rations were prepared at the poultry farm. Feed and water were supplied ad libitum. The amount of feed offered to each group of hens and the amount refused was weighed and recorded every 28 days (one period) in order to determine feed-intake. Daily records of egg production were kept except for the first five days in order that the animals could get accustomed to the new diets. During the last three consecutive days of each period eggs were weighed and broken on a glass plate for direct measuring of the Haugh units. The birds were weighed individually at the beginning and at the termination of the experiment. Records of mortality were maintained. Statistical Analysis All data were analyzed by analysis of variance, orthogonal polynomials, using General Linear Model (GLM), Quadratic Model sub- routine of Statistical Analysis System (SAS). Significant differ- ences between means of treatment, overall means of supplemented and unsupplemented hydrolyzed feather meal rations were tested at the level of 5 percent (Appendix, Tables 14 through 20) by the use of Split-Plot repeat measurement procedure (Snedecor and Cochran, 1967). CHAPTER V RESULTS AND DISCUSSION The effects of various treatments and/or lysine and methionine supplements on performance are shown in Table 7. Production The percent production did not show a significant difference between the means of treated and control groups (Appendix, Table 14). Work by Leveille and Fisher (1960) showed diets at the level of 7 per- cent HFM (Hydrolyzed Feather Meal) with supplemental methionine were capable of supplying the estimated minimal essential amino acids, but it was unable to support maximal production. In the present experiment the percentage of lysine and methionine became lower than the NRC requirement for the laying hens only at the level of 9 percent HFM with no supplementation of amino acids (Table 6). There was a sig- nificant (P<:0.05) difference between the percentage of egg production of the birds receiving the 3 percent hydrolyzed feather meal diet with no supplementation of methionine and lysine (HFM-) and those receiving the 9 percent HFM- diet (Appendix, Table 14). Figure 2 shows that the percentage of production was generally higher in the hens that were fed the diet with 3 percent HFM+ than in the control group. 25 26 .mupom ocean mo cowuuucwempqaam 0: new: Fame emnummw umuxpocu»: .mmmm oNP mo mmacm>m mg» mp Sauce scum use mewmaFV mumum ocean mo comumucmEmpnqam gap: Fame Locummm um~apocuaz 1zmzu .Amepcopsume +=e=u a .Amma mo mxmmz ow o» om. muomcma upon: new comuuanoea mo unaccomm e.m~_ o._m o.o~. m.o- we.mm ma.~m 1:.1 am _.mmm m.mm m.-_ 0.55. .m.em me.mm 12;: an o.m- A.om e..m_ m.m~. me.om m..m~ u-zex an m.e_m ..mm ~.mm_ m.om~ m~.em me.mm +z.= em m.m~m ..mm «.mm_ o.~e~ mm.mm mm.mm +2.: we o.¢o~ o.~m e.Fm_ e._m. m_.em mm.ao u+2ez Rm m.e- o.mm _..N_ m.mmp m~.mm me.~m Poeueou when oe.\u..m b_e= swo\=m:\ mmu\ b;m_a= mam gawbuaeoe. moeaeoaaee \Amsocmv :Poo agave: zone: a a a Amsmcmv compasamcou ummu mew: memxe. $0 museELomcma co mumps Apewz cmzuemd um~zpogvxzv an: o» mumo< ocmE< —mwucmmmm mo mezuxm: use mcwm». .mcwcowgumz cum: :o_uou:m5opaa=m xgmummo we uommeu use N m—noh Production Percent 27 35' .—o 3% HFM+ H 60/. HFM+ A___A 97o HFM+ 30. c. ....... n CONTROL 175' o R. 70" I‘ 625L so» 55' 50' e Period Figure 2. Percent Production Response by Period for Control and Supplemented Groups. 28 There was no significant difference in percent production of hens fed the diets supplemented with HFM+ (Appendix, Table 14). Egg production of the hens which were fed the diets with 3 and 6 percent HFM, whether supplemented or unsupplemented, improved when the amino acid requirement was not lower than that of National Research Council (Figures 2 and 3). Production was higher at 3 than at 6 percent HFM. Peak production occurred in all treated groups at an earlier age than in the control birds (Table 8 and Figures 2 and 3). Thus, there may be some factor or factors in the hydrolyzed feather meal (HFM) which stimulated the birds to peak earlier. Figure 4 shows that increasing the percentage of HFM in the diet of laying hens linearly decreased the percentage of production. This reduction of production by additional HFM was statistically significant (P<:0.05) for the hens that were fed rations with no supplementation of lysine and methionine (Appendix, Table 14). The percentage of production significantly (P<:0.05) changed by periods. Peak production was achieved between 24 and 28 weeks of age for all test groups and between 28 and 32 weeks of age for the control group (Figures 2 and 3). Egg Weight There was no significant difference between weights of eggs produced by treated hens and those produced by control hens (Appendix, Table 15). Van and Payne (1977), in comparing a control diet with one containing 7 percent HFM, showed that as supplementation of lysine 29 .——-.3% HFM- 85r- o——06% HFM- u -------- :1 Control 80 - 11 75 - 1o - 0”". ‘1, Percent Produciion 0| 0* 0 0| 0 0| I . I 50 45 Po rlod Figure 3. Percent of Production Response by Period for Control and Unsupplemented Groups. 30 Table 8 The Percentage of Production in the Control Group, Supplemented and Unsupplemented Groups Periodsa Treatments lst 2nd 3rd 4th 5th Controlb 59.8 59.9 71.57 55.5 55.9 Average of supplementedc 52.1 75.1 55.4 59.9 58.46 Average of unsupplementedc 57.7 77.7 72.5 52.5 55.0 aEach period is 28 days long. 24 birds. bThe percentage of production in this row is obtained from cThe percentage of production in these rows are obtained from 72 birds. pe rcenl producllon 31 ‘ O——o H m- Hurm‘ 0 Control 0 3 6 D D Perceni OI HFM Figure 4. Percent of Production Response to HFM. 32 increased egg weight improved. This was supported by the present experiment (Figure 5), but it was not true when no supplementation was added. Figure 5 shows that the egg weights were suppressed when no supplementation of lysine and methionine were put into the diets. Moran et a1. (1969) showed the increase in egg weight resulting from the addition of amino acids into the diets containing 5 percent HFM. Egg weight from all groups of birds increased throughout the duration of the experiment. However the egg weight increased signif- icantly more (P<:0.05) in the hens which were fed diets with lysine and methionine supplementation than in those with no supplementation of amino acids (Appendix, Table 15 and Figure 6). Table 9 shows the average egg weight for each period. The average egg weight was sig- nificantly (P&:0.05) less in the hens which were fed with 9 percent HFM- than in those fed the 3 percent HFM- diet (Appendix, Table 15). Also the average egg weight from the hens which were fed the diets with HFM- (with no supplementation of amino acids) was significantly lower (P<:0.05) than that of eggs from the supplemented groups (Appendix, Table 15). Feed Consumption and Nutrient Intake Feed consumption by periods is shown in Table 10. There were no significant differences in feed consumption due to treatments (Appendix, Table 16). Figure 7 shows the relationship between feed consumption and percentage of HFM. Moran et al. (1969) demonstrated an improvement in feed-intake by adding 5 percent HFM while maintaining the lysine and methionine AVERAGE EGG WEIGHTIQroms/egg) 57 56 54 53 f U 33 .——.HFM- ‘ Hun-1+ 5 5 PERCENT OF H FM Figure 5. Egg Weight Response to HFM in Diets. AVERAGE EGG wslowusromxegg) 59 57 55 53 51 1 34 HHFM- ‘ H HFM+ a ooooo 00:] CONTROL - 2 £- 4‘ PERIOD Figure 6. Egg Weight Response to Periods. Table 9 35 The Average of Egg Weight in the Control Group, Supplemented and Unsupplemented Groups Periodsa Treatments lst 2nd 3rd 4th 5th Controlb 51.8 54.3 55.9 57.3 58.5 Average of supplementedC 51.8 54.8 57.0 58.4 58.8 Average of unsupplementedc 51.3 52.8 55.5 55.5 57.4 aEach period is 28 days. bThe average of egg weight in this row obtained from 72 eggs (grams/e99). cEach datum in these rows is the average of 216 eggs (grams/e99). The Average of Feed Consumption in the Control Group, Supplemented and Unsupplemented Groups 36 Table 10 (grams/hen/day) Periodsa Treatments lst 2nd 3rd 4th 5th Controlb 113.7 125.0 125.8 114.4 125.2 Average of supplementedc 111.5 129.9 128.9 134.0 139.0 Average of unsupplementedc 102.5 129.4 132.0 128.5 125.5 aEach period is 28 days. b day). Each datum is the average of 24 birds consumption (grams/hen/ cEach datum in these rows is the average of 72 birds consumption (grams/hen/day). 37 requirement for laying hens in the diets. In this study there was not any significant (P>»0.05) change of feed-intake in the treated groups whether it is supplemented or not (Appendix, Table 16). The level of protein and age of birds used may explain this difference. The feed-intake significantly (P<:0.05) changed by periods (Appendix, Table 16). Figure 8 shows the relationship between feed consumption and periods in the treated and control groups. Table 11 shows that there was little difference between calories and proteins intake among the treatments. Lysine and methionine intake varied from 1.138 and 0.375 to 0.495 and 0.242 grams/hen/day, respectively. Feed Conversion Feed efficiency for birds on each treatment is shown in Figure 9. There was no significant (P>10.05) difference in feed conversion between the birds receiving the supplemented rations (HFM+) and those receiving the unsupplemented rations (HFM-). The results obtained with 3 and 6 percent HFM with and without supplementation of amino acids did not improve the feed efficiency (Appendix, Table 17 and Figure 9). However, the level of lysine and methionine matched the NRC requirement for the laying hens. Moran et a1. (1969) showed the improvement of feed efficiency in the hens that were fed at 5 percent HFM with supplementation of amino acids. Van and Payne (1977) carried out an experiment which showed the improvement of feed efficiency at the level of 7 percent HFM into the ration of laying hens when the amino acids were supplied to the requirement level of laying hens. These variations of results 38 .me\=m;\mEmcw a .xmc\=mg\mvcopmum en.m mm.m 5F.e oo.e mp.¢ op.e mm.m asapopou mm_.o oo~.o mmm.o mom.o mm~.o mmN.o w-.o neecaouaxgp NvN.o som.o Fem.o Nmm.o mpe.o noe.o mum.o ago-oeweomgpm: mm¢.o nmm.o Fpo._ emw.o mpm.o ._mo.p mm—.P nmcwmxo e¢.mp mm.o~ oe.- e~.mm nm.m~ mo.m~ mN.P~ acmmuoca muzcu m.mem o.—mm N.nom F.5om ¢.pmm N.o~m m.mem mwmcopnu 12;: gm 12¢: no -2m: gm +ze: no +ze: No +2e: mm Focpcou Hemmeuaz Auv :owumm Amy compmm Amy comumm on compem Auv cowumm Amy cowpmm A’0.05) between the control 53 group and the birds that received the 3, 6 and 9 percent HFM+ (hydrolyzed feather meal with supplementation of amino acid) rations in egg production and egg weight. Egg weight in all treated and control groups was significantly increased (P<:0.05) by the age of the birds. Feed-intake, feed conversion Haugh units and mortality did not significantly change (P> 0.05) between the control and treated groups. All the above factors except mortality significantly changed (P<:0.05) by periods in various treatments. The overall weight gain in 140 days of the experiment was significantly different between the various treatments. 0n the basis of this study it can be concluded that the amino acid requirement of laying hens can be met when up to 6 percent hydrolyzed feather meal is included in a corn-soybean type diet. Inclusion of 9 percent HFM in the diet has deleterious effects on production and egg weight if the diet is not supplemented with lysine and methionine. APPENDIX 54 Table 14 Analysis of Variance by the Use of Split-Plot Repeat Measurement for Egg Production (%) Source of Sum of Mean Variation df Square Square F P>1F Treatment 6 377.22 62.87 1.94 0.05 B vs. 0a 1 58.6 -- 1.81 0.05 E vs. Gb 1 241.3 -- 7.46 < 0.05* Bi-D vs. C 1 26.7 -- 0.826 0.05 Ei-G vs. F 1 35.3 -- 1.09 0.05 Control vs. treated 1 1.22 -- 0.038 0.05 Supplemented vs. unsupplemented 1 13.75 -- 0.425 0.05 Error a 14 452.662 32.33 -- -- Period 4 489.425 122.356 26.848 < 0.05* Period by ‘ treatment 24 114.845 4.785 1.05 0.05 Error 0 56 255.212 4.56 -- ~- Tbtal 104 2066.234 *These contrasts are significantly different. aB, C and D diets are 3, 6'and 9 percent hydrolyzed feather meal, respectively, with amino acid supplementation. bE, F and G diets are 3, 6 and 9 percent hydrolyzed feather meal, respectively. without supplementation of amino acid. 55 Table 15 Analysis of Variance by the Use of Split-Plot Repeat Measurement for Egg Weight (grams/egg) Source of Sum of Mean Variation df Square Square F P>1F Treatment 6 129.59 21.599 2.356 0.05 B vs. 0a 1 2.44 -- 0.266 0.05 E vs. Gb 1 67.17 -- 7.330 <0.05* Bi-D vs. C 1 8.76 -- 0.955 0.05 Ei-G vs. F 1 4.18 -- 0.456 0.05 Control vs. treated 1 1.38 -- 0.150 0.05 Supplemented vs. unsupplemented l 46.46 -- 5.068 < 0.05* Error a 14 128.34 9.167 -- -- Period 4 620.898 155.224 126.93 <:0.05* Period by ‘ treatment 24 40.597 1.692 1.383 0.05 Error b 56 68.48 1.22 -- -- Total 104 1118.296 *These contrasts are significantly different. 38, C and D diets are 3, 6 and 9 percent hydrolyzed feather meal, respectively, with supplementation of amino acid. b meal, respectively without supplementation of amino acid. E, F and G diets are 3, 6 and 9 percent hydrolyzed feather 56 Table 16 Analysis of Variance by the Use of Split-Plot Repeat Measurement for Feed Consumption (kilograms/period) Source of Sum of Mean Variation df Square Square F P>1F Treatment 6 1.120 0.1867 0.904 0.05 B vs. 06‘ 1 0.0136 -- 0.065 0.05 E vs. Gb 1 0.380 -- 1.840 0.05 Bi-D vs. C 1 . 0.011 -- 0.053 0.05 Ei-G vs. F 1 0.014 -- 0.067 0.05 Control vs. treated 1 0.253 -- 1.225 0.05 Supplemented vs. unsupplemented 1 0.46 -- 2.22 0.05 Error a 14 2.89 0.2065 -- -- Period 4 6.339 1.585 31.41 ‘<0.05* Period by. ‘ treatment 24 2.254 0.094 1.86 0.05 Error b 56 2.825 0.050 -- -- TPta‘ 104 16.559 *This contrast is significantly different. 38, C and 0 diets are 3, 6 and 9 percent hydrolyzed feather meal, respectively, with supplementation of amino acid. b meal, respectively, without supplementation of amino acid. E, F and G diets are 3, 6 and 9 percent hydrolyzed feather 57 Table 17 Analysis of Variance by the Use of Split-Plot Repeat Measurement for Feed Conversion (grams of feed/egg) Source of Sum of Mean Variation df Square Square F P>1F Treatment 6 67062.3 11177.0 1.256 0.05 B vs. 0a 1 15833.2 -— 1.779 0.05 E vs. 0” 1 23129.6 -- 2.660 0.05 8+-0 vs. 0 1 752.3 -- 0.084 0.05 Ei-G vs. F 1 993.14 -- 0.111 0.05 Control vs., treated 1 17411.14 -- 1.950 0.05 Supplemented vs. unsupplemented 1 17787.26 -- 2.009 0.05 Error a 14 124592.97 -- -- -- Period 4 55186.5 13796.6 8.530 <:0.05* Treatment by period 24 40940.65 1705.86 1.050 0.05 Error b ' 56 90523.98 1617.42 -- -- Total 104 453319.9 *This contrast is significantly different. aB, C and D diets are 3, 6 and 9 percent hydrolyzed feather meal, respectively, with supplementation of amino acid. bE, F and G diets are 3, 6 and 9 percent hydrolyzed feather meal, respectively, without supplementation of amino acid. 58 Table 18 Analysis of Variance by the Use of Split-Plot, Repeat Measurement for Haugh Units Source of Sum of Mean Variation df Square Square F P>1F Treatment 6 552.30 92.05 1.43 0.05 B vs. 0" 1 16.72 -- 0.259 0.05 E vs. Gb 1 137.80 -- 0.140 0.05 Bi-D vs. C 1 4.62 -- 0.072 0.05 Ei-G vs. F l 4.58 -- 0.071 0.05 Control vs. treated 1 8.11 -- 0.126 0.05 Supplemented vs. unsupplemented 1 4.40 -- 0.069 0.05 Error a 14 901.29 64.38 -- -- Period . 4 1070.30 267.58 3.480 < 0.05* Period by . treatment 24 1883.40 78.47 1.020 0.05 Error b 56 4301.7 76.90 -- -- Total 104 8885.22 *This contrast is significantly different. aB, C and D diets are 3, 6 and 9 percent hydrolyzed feather meal, respectively, with supplementation of amino acid. b meal, respectively, without supplementation of amino acid. E, F and G diets are 3, 6 and 9 percent hydrolyzed feather 59 Table 19 Analysis of Variance by the Use of Split-Plot Repeat Measurement for Mortality Source of Sum of Mean Variation df Square Square F P>aF Treatment 6 0.857 0.1428 0.940 0.05 B vs. 0a 1 0.300 -- 1.969 0.05 E vs. 0” 1 0.300 -- 1.970 0.05 84-0 vs. C 1 0.011 -- 0.072 0.05 Ei-G vs. F 1 0.011 -- 0.072 0.05 Control vs. treated 1 0.057 -- 0.374 0.05 Supplemented vs. unsupplemented 1 0.177 -- 1.160 0.05 Error a 14 2.133 0.1524 -- -- Period 4 0.1524 0.038 0.271 0.05 Period by treatment 24 2.381 0.0992 0.706 0.05 Error b 56 7.866 0.1405 -- -- TPtal 104 14.2454 No contrasts are significantly different. aB, C and D diets are 3, 6 and 9 percent hydrolyzed feather meal, respectively, with supplementation of amino acid. b E, F and G diets are 3, 6 and 9 percent hydrolyzed feather meal, respectively, without supplementation of amino acid. 60 Table 20 Analysis of Variance by the Use of Split-Plot Repeat Measurement for Average of Weight Gain Source of Sum of Mean Variation df Square Square F P>1F Main effect 6 123017.7 20502.9 3.276 < 0.05* Treatment 6 123017.7 20502.9 3.276 < 0.05* Explained 6 123017.7 20502.9 3.276 < 0.05* Residual 14 87628.0 6259.1 -- -- Total 20 210645.8 10532.3 *Significantly different. 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