METABOLISM AND NUTRITION. Glandless Cottonseed Meal for Laying and Breeding Hens and Broiler Chicks

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1 METABOLISM AND NUTRITION Glandless Cottonseed Meal for Laying and Breeding Hens and Broiler Chicks J. R. RYAN, F. H. KRATZER, 1 C. R. GRAU, and P. VOHRA Department of Avian Sciences, University of California, Davis, California 916 (Received for publication April 29, 1985) ABSTRACT The Acala C-9 variety of glandless cottonseed meal (GCSM) was tested for available gossypol in a 1-day feeding trial with laying hens. Corn-soybean diets incorporating GCSM at 1 and 2% were compared with diets formulated with Acala SJ-5, a commercially available cottonseed meal (CSM), at 1% and a control diet with no CSM. Each diet was fed to 25 hens. No detectable gossypol was found in the yolks of eggs from hens fed GCSM or the control diet, but gossypol was present when hens were fed the diet with CSM. Egg yolk discoloration in an ammonia atmosphere was noted in all of the eggs tested from the CSM-fed hens but only at a very low incidence in eggs from other groups. There were no differences between any of the four treatments in percent egg production, average egg weight, or hatchabihty. Broiler chicks were fed for 19 days diets with GCSM as the sole source of protein to determine the limiting amino acids. The diets contained 23% protein, 32 kcal metabolizable energy/kg, and National Research Council levels of vitamins and minerals. Diets were supplemented with lysine, methionine, leucine, and isoleucine from to % and were kept isonitrogenous with glutamic acid. Only lysine was limiting in GCSM for chick growth. Chick weights did not differ significantly from those fed an adequate reference diet if % lysine was added. (Key words: glandless cottonseed meal, protein source, egg production, hatchabihty, yolk discoloration, gossypol) INTRODUCTION Cottonseed meal (CSM) is used extensively in diets for chickens, turkeys, swine, and ruminants but not for laying hens, because of its potential deleterious effect on egg quality. Schaible et al. (1934) showed that the gossypol present in CSM fed to hens is responsible for the marked discoloration of yolks that occurs during the storage of eggs. Grau et al. (1954) and Woronick and Grau (1955) found that when gossypol is fed to laying hens, a yellow component, which is different from gossypol and is presumably a gossypol-cephalin complex, is present in the egg yolk. Heywang et al. (1955) reported that the percentage of discolored yolks and the degree of discoloration increased during storage of eggs as the level of free gossypol in the diet increased. Heywang et al. (1949) observed reduced hatchabihty when 2 or 3%, but not 1%, of hydraulic or solvent processed CSM was present in the diet. In further studies, egg production and feed consumption were reduced when the 1 To whom correspondence should be addressed Poultry Science 65: free gossypol content of the diet was.2% or higher (Heywang and Bird, 1954). Narain et al. (1957) added cottonseed pigment glands to a laying hen diet and found that.2% free gossypol depressed feed consumption, and higher levels caused reduced body weight, egg production, and egg size. El-Abbady et al. (1975) reported that.18% and.27% free gossypol in the diet of hens depressed hatchabihty, but free gossypol up to 2% had no effect on egg production or egg weight. According to Lyman et al, (1953), gossypol is an important factor in influencing the nutritional value of cottonseed protein. However, lysine supplementation of diets containing CSM with low levels of free gossypol resulted in good growth of chickens. Anderson and Warnick (1966) determined that lysine is the most limiting amino acid in cottonseed protein for chickens. Because gossypol is contained in the pigment glands of the cottonseed, the development of a strain of cotton in which the seeds are without pigment glands and, therefore, low in gossypol should eliminate the problems associated with feeding CSM. When half or all of the soybean Downloaded from at Penn State University (Paterno Lib) on March 6,

2 95 RYAN ET AL. meal in a laying diet was replaced by glandless cottonseed meal (GCSM), some yolk discoloration in eggs was observed (Heywang and Vavich, 1965a,b), but only a few egg yolks were discolored after 6 months of storage. Almost all of the eggs showed discoloration when regular cottonseed meal was used. Egg production and hatchability were about the same with both meals. Anderson and Warnick (1966) found more available lysine in GCSM than in regular meal; however, methionine and lysine were equally limiting. Waldroup et al. (1968) reported that lysine supplementation to the ration was only needed when 75% or more of the soybean meal in a broiler diet was replaced by GCSM. Apparent lysine availability was 62.1% for GCSM and 49.7% for regular cottonseed meal (Reid et al, 1984). Although glandless cottonseed strains have been available for several years, they have not been used significantly for commercial cotton production because of poor yield and fiber quality characteristics. A strain developed by H. B. Cooper at the University of California Cotton Experiment Station at Shafter, CA, TABLE 1. Composition 1 Constituent Crude protein Moisture Fat Crude fiber Ash Lysine, total Lysine, available Arginine Threonine Glycine Serine Cystine Methionine Valine Isoleucine Leucine Tyrosine Phenylalanine Gossypol, free Gossypol, total overcame these drawbacks and was selected for commercial use. It was grown in large enough quantities to test its suitability in poultry diets. The effects of the GCSM were compared with those of a regular CSM in a laying diet on egg production, hatchability, and on egg yolk discoloration. Glandless cottonseed meal was also tested for its suitability as a protein source for chickens with and without supplementation with various amino acids. MATERIALS AND METHODS Glandless cottonseed of an experimental Acala C-9 strain was processed commercially by the prepress solvent method to produce the GCSM used in these studies. The composition of GCSM along with that of a commercially processed CSM that was used in a control diet is shown in Table 1. The metabolizable energy (ME) of the GCSM as determined by a rapid method with adult male chickens (Vohraeia/., 1982) was 17 kcal/kg. This value, along with those in Table 1, were used in the computerized formulation of diets. of glandless and glanded cottonseed meals Glandless cottonseed meal <.1 4 (/o) - Glanded cottonseed meal Downloaded from at Penn State University (Paterno Lib) on March 6, 216 'Analyses by Woodson-Tenent Labs, Inc., Memphis, TN, according to Association of Official Analytical Chemists (1984) methods. Amino acids determined after acid hydrolysis. 2 Analyses, except gossypol, by ABC Labs, Inc., Columbia, MO.

3 GLANDLESS COTTONSEED FOR LAYERS AND BROILERS 951 The diets for the laying hen trial were formulated to contain 17% protein and 28 kcal ME/kg (Table 2). The control diet was based on corn and soybean meal and the test diets contained 1 or 2% GCSM or 1% CSM. All diets were formulated to meet the National Research Council (NRC, 1977) recommended levels for nutrients. Laying Hen Studies. Hy-Line White Leghorn hens, approximately 3 weeks of age and in their first laying cycle, were randomly selected into groups of 25 hens each, weighed, and fed the experimental diets. The hens were kept in individual cages in an open-sided house and were provided feed and water ad libitum. A lighting schedule of 16L:8D was maintained for the 1-day duration of the experiment. Eggs were selected at random during the last 5 days of the experiment for gossypol determination. The yolks were extracted with acetone, followed by hexane-acetone (3:1) according to the method of Grau et al. (1954). For the study to determine the tendency for yolks to discolor, 11 eggs per treatment were broken out within a day after lay during the last 3 days of the trial. Ingredients Corn Soybean meal (47% protein by analysis) Limestone Alfalfa meal (17% protein) Dicalcium phosphate, dihydrate Premix 2 NaCl DL-Methionine Wheat bran Soybean oil GCSM CSM Total Eggs were saved for incubation during the last 3 days of the trial for the fertility and hatchability study. The hens were inseminated on the 1st and 3rd days of the week prior to saving eggs and once a week thereafter. Pooled semen from males fed a conventional ration without CSM was used for insemination. Eggs were collected and weighed daily and production was recorded for individual hens. Broiler Chick Studies. For the broiler growth studies, the basal diet was formulated to provide 23% protein and 32 kcal ME/kg using GCSM as the sole source of protein (Table 3). From the amino acid profile, GCSM protein is deficient in isoleucine, leucine, lysine, and methionine (Table 4). In the feeding trial, the basal diet containing GCSM was supplemented with these amino acids. In addition, lysine and methionine were supplemented at levels of.1,, and % to determine the degree of deficiency of these amino acids. Diets were kept isonitrogenous by the addition of glutamic acid. A commercial chick starting diet based largely upon corn and soybean meal was used as an adequate reference diet. TABLE 2. Percentage composition of diets for laying hens 1 Control Day-old male broiler chicks were randomly 1% GCSM % GCSM % CSM Downloaded from at Penn State University (Paterno Lib) on March 6, 216 Calculated analyses Metabolizable energy, kcal/kg Protein, % Ca, % Available P, % GCSM = Glandless cottonseed meal, CSM = cottonseed meal. 2 Supplied the following levels per kilogram of diet: vitamin A, 9 IU; vitamin D, 15 ICU; vitamin E, 11 IU; vitamin K, 1 mg; choline, 52 mg; riboflavin, 6 mg; pantothenic acid, 8 mg; vitamin B 12,.12 mg; niacin, 24 mg; folacin, 1 mg; biotin, mg; Mn, 73 mg; Zn, 6 mg; Cu, 5 mg.

4 952 RYAN ET AL. Ingredients TABLE 3. Composition of chick diet using glandless cottonseed meal as the sole protein source (%) wire floors. The experiment was continued for 14 days. The significance of the growth data was determined by analysis of variance and by the method of Duncan (1955). Glandless cottonseed meal (GCSM) Corn starch Soybean oil Dicalcium phosphate, dihydrate Limestone NaCl Premix 1 Total Calculated analysis Metabolizable energy, kcal/kg Protein, % Ca, % Available P, % Amino acid Arginine Glycine + serine Histidine Isoleucine Leucine Lysine, total Lysine, available Methionine Phenylalanine Threonine Tryptophan Valine RESULTS AND DISCUSSION The lower egg production in the control group without CSM in comparison with the other groups (Table 5) was due to a decline in egg production during the 3rd and 4th weeks of the experiment. A rather dramatic decrease in egg production for this group was corrected by the addition of.1% DL-methionine to the diet. By the end of the experiment, egg production per hen per day was equal to the other groups, but the number of eggs for the total period was significantly lower. The methionine content of the control diet was 1% (by calculation), which was greater than the requirement for laying hens (.27%) given by the 1977 NRC table but lower than the 2% given in the 1984 revision. Average egg weight, percent hatchability of fertile eggs, and average chick weight were not significantly (P<.5) different in any of the treatments. Fifty-six eggs from each treatment were broken out into petri dishes and placed in a desiccator with an atmosphere of ammonia according to the method of Heywang et al. (1961). Two eggs from the control diet and six from the group fed 2% GCSM showed slight TABLE 4. Amino acid composition of chick diets with glandless cottonseed meal (GCSM) compared with amino acid requirement Dietary requirement %NRC' Supplied the following levels per kilogram of diet: vitamin A, 585 IU; vitamin D, 12 ICU; vitamin E, 1.6 IU; vitamin K, 4 mg; thiamin, 8.9 mg; riboflavin, 1 mg; pyridoxine, 8.2 mg; vitamin B 12,.1 mg; choline, 13 mg; niacin, 1 mg; pantothenic acid, 3 mg; biotin, mg; folic acid, 2 mg; Mn, 81.2 mg; Mg, 66 mg; Zn,.2 mg; Fe, 8 mg; Cu, 4.1 mg; Mo, 2 mg; Co, 4.7 mg; Se,.1 mg; I, 1 mg. distributed in duplicate groups of 1 each for each treatment, weighed, wing banded, and fed the diets ad libitum. The chicks were housed in electrically heated battery brooders with raised % Present GCSM Diet %of NRC requirement Downloaded from at Penn State University (Paterno Lib) on March 6, 216 'NRC = National Research Council.

5 GLANDLESS COTTONSEED FOR LAYERS AND BROILERS 953 TABLE 5. Effect of feeding diets containing glandless (GCSM) and glanded cottonseed meal (CSM) to bens on egg production, egg weight, hatchability, and chick weight Number Hatchaof eggs Avg bility of Avg Dietary for 1 egg fertile chick treatment days weight eggs weight (g) (%) (g) Control % GCSM % GCSM % CSM yolk discoloration (Table 6), possibly from causes unrelated to CSM. In contrast, all eggs from the group fed 1% CSM showed distinct yolk discoloration in the ammonia atmosphere. The absorption spectra of the yolks from the control and GCSM-fed hens showed no indication of the gossypol-cephalin complex (Fig. 1) whereas the egg yolks from the hens fed 1% of the GCSM showed the characteristic spectrum of the complex. The lack of an effect of either 1 or 2% GCSM or 1% CSM upon egg production or hatchability agrees with the reports of Heywang et al. (1949), Heywang and Bird (1954), and Narain et al. (1957) in which 1% of conventional CSM or free gossypol levels below.2% caused no effect on egg production or hatchability. El-Abbady et al. (1975) reported reduced hatchability with.18% free gossypol. The free gossypol levels from the GCSM and CSM diets were less than.2 and.47%, respectively. Reid et al. (1984) observed a slight decrease in egg production with feeding 15% of TABLE 6. Yolk discoloration after subjecting eggs to an atmosphere of ammonia GCSM, whereas 5 and 1% caused no effect on egg production. Vohra et al. (1975) obtained satisfactory production with 1 and 15% CSM in laying rations, although there was some yolk discoloration. In the current study, the free gossypol was sufficiently high so that egg yolk discoloration was noted when egg yolks from hens fed 1% CSM were placed in an ammonia atmosphere. Yolks from hens fed 1% of GCSM showed no discoloration and even feeding 2% caused only a low incidence of discoloration. The lack of an effect of the GCSM on yolk discoloration was also shown by absence of the spectrum characteristic of the gossypol-cephalin complex in eggs from hens fed as much as 2% of this meal. 1.6 i: : Downloaded from at Penn State University (Paterno Lib) on March 6, 216 Dietary treatment 1 Total number of eggs treated Number of yolk discolored Control 1% GCSM 2% GCSM 1% CSM GCSM = Glandless cottonseed meal, CSM = cottonseed meal WAVELENGTH - mjj FIG. 1. Absorption spectra of hexane-acetone (3:1) extracts of egg yolks of hens fed control diet or diets containing glandless (GCSM) or glanded cottonseed meal (CSM).

6 954 RYAN ET AL. TABLE 7. Effect of amino acid supplementation on growth and feed efficiency (gain/feed) of chicks fed a diet containing glandless cottonseed meal as the sole source of protein Lysine % Added L-amino acids Methionine Isoleucine Leucine Glutamic acid Average body weight gain Feed efficiency.1 Stock mash Means with the same superscript do not differ significantly (P<.5). The growth of chicks fed GCSM as the only source of protein was not influenced by the addition of glutamic acid (Table 7). There was no response to the addition of methionine, isoleucine, and leucine in the absence of lysine. The addition of lysine with the other three amino acids caused a significant growth improvement with the best response occurring at %. Omission of leucine or isoleucine caused no reduction in growth, but the omission of methionine resulted in a slight numerical reduction in growth that was not statistically significant. Growth of all of the groups supplemented with % or more lysine, regardless of other amino acid supplementation, was not statistically different from that of chicks fed a practical starting diet. Feed efficiency was also improved by the addition of lysine and was not influenced by the omission of other amino acids. These results agree with the reports of Lyman et al. (1953) and Anderson and Warnick (1966) that lysine is the most limiting amino acid in cottonseed protein. The fact that leucine and isoleucine were not limiting and that these amino acids are usually well supplied by practical dietary ingredients indicate that they are not of concern when using GCSM in practical diets. Methionine probably should (g) 155<kf 151* 137 f 168 cde 176bcde 26 ab 19 abc 25 ab 21* 26 ab 22 a 22 ab 24 ab be considered in formulating diets with this feedstuff. The results of this study show that the gossypol level in cottonseed meal produced from the Acala C-9 strain of glandless cotton seed is sufficiently low to cause no problems in egg production, hatchability, or egg yolk discoloration in laying hens. Lysine is the most limiting amino acid when it is used as the sole source of protein in a diet for broiler chicks. ACKNOWLEDGMENTS We are grateful to Cotton Incorporated, Raleigh, NC, for providing financial support for this project. The experimental cottonseed meal was supplied by California Planting Cottonseed Distributors, Bakersfield, CA; J. G. Boswell Company, Corcoran, CA; and Ranchers Cotton Oil Company, Fresno, CA. Crude soybean oil was kindly supplied by Central Soya, Decatur, IN. REFERENCES Anderson, J. O., and R. E. Warnick, Sequence in which essential amino acids become limiting for growth of chicks fed rations containing cottonseed meal. Poultry Sci. 45: Association of Official Analytical Chemists, Downloaded from at Penn State University (Paterno Lib) on March 6, 216

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