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CENTRO INTERNACIONAL DE ALTOS ESTUDIOS AGRONÓMICOS MEDITERRÁNEOS INSTITUTO AGRONÓMICO MEDITERRÁNEO DE ZARAGOZA INFLUENCE OF THE MAIN CEREAL AND FEED FORM OF DIET ON PRODUCTIVE PERFORMANCE AND DIGESTIVE TRACT TRAITS IN BROWN-EGG LAYING PULLETS FROM HATCHING TO 17 WEEK Ramzi Harzalli Trabajo realizado en el Departamento de Producción Animal, Escuela Técnica Superior de Ingenieros Agrónomos, Universidad Politécnica de Madrid, bajo la dirección del Dr. Gonzalo González MATEOS Y presentado en lectura pública el día 9 de julio de 2013 ante el siguiente tribunal: Se le ciegan los ojos con el polvo, Oyendo siempre la canción del tiempo Recuerda, caminando en campo solo, Nos pusieron descalzos en la tierra Quemarnos el dolor, pero algo así, Como un dolor sin sitio destinado La mañana no siempre nos descubre Tras el vocablo, el mito o el ensueño He aprendido que la vida es dura Pero yo lo soy más! AGRADECIMIENTOS Agradezco a dios, por ser mi continua fortaleza en cada meta que me propongo e iluminado mi pensamiento y en especial por darme valor en los momentos más difíciles de mi vida. A mi querida familia mi refugio, que siempre me ha apoyo y animado. Mi madre Fatma Baccouch, mi abuela Sabeha Benit Mouldi elkefi, mi abuelo Majid Baccouch ejemplos de determinación, paciencia, amabilidad y confianza. Mis hermanos Sondes y Ali Harzalli, y demás familiares gracias por su eterno cariño, oraciones y ayuda que han significado mucho para mí. Agradezco la buena disposición de todo el personal de la administración del Escula superior de agricultura del ELkef (ESAK). Un agradecimiento especial para Mokhtar Mehouichi, Bea ben harrath, Mathlouthi Najib y Ridha bergaoui. Agradezco la buena disposición de todo el personal de la administración del Instituto agrónomo mediterráneo Zaragoza (IAMZ) y Universidad Politécnica Madrid (UPM) que me ayudó mientras realizaba el trabajo de investigación. Un agradecimiento especial para Armando Occon, coordinador del Curso de Producción Animal. También a Ramzi Bel Khouja por su disponibilidad continúa para ayudarme. Un agradecimiento especial a mi Director de Tesis, Dr. Gonzalo González Mateos por darme el privilegio de incorporarme en su grupo de investigación, por enseñarme y por sus oportunos consejos y correcciones y por darme la oportunidad de ampliar mis conocimientos en el campo de la nutrición en avicultura. Su apoyo, rigor, dedicación y disponibilidad me han permitido acabar este trabajo con mucho aprovechamiento. Miles de gracias. A mis amigos Husham Mandalawi, Mohamed frikha, Rodriguo Nourhan El Abed, Vahid Mohamed y Ridha Ibidi han estado dispuesta a ayudarme en lo que sea y a la hora que sea. Todos los profesores del Departamento de Producción Animal de UPM, por su entera disponibilidad y colaboración prestada a todas mis preguntas y dudas durante en el transcurso de mi estancia en el Departamento. A la empresa Cantos agroalimentaria por aceptar mi estancia durante el desarrollo del trabajo experimental. A todo el personal de la empressa y especialmente a Adriano Pérez por su gran contribución y colaboración en la realización de los trabajos experimentales, su constante apoyo, su gran ayuda y sobre todo por esa gran amabilidad. A mis amigos Sofien Egtari, Ahmad Hamemi, Khawla Samaali, Nedia Yacoubi, Foued Bouazizi, Khawla ELHabessi, Fathi Ayari, y Carlos que saben que desde lo más profundo de mi corazón les agradezco el haberme brindado todo el apoyo, ayuda, cariño y amistad y por estar siempre pendientes de mí, sobre todo, en aquellos momentos en los que más los he necesitado. A los compañeros y becarios del Departamento, Nuria, Lourdes, Jorge, Julio, Bea y Pilar, quiero agradecerles el buen trato que siempre me han dispensado y sobre todo con esa especial amabilidad y buen humor, durante el transcurso de mi estancia. INDEX Index Chapter 1: Abstract 1.1. ABSTRACT ..................................................................................................... ii 1.2. Resumen .......................................................................................................... iii 1.3. Résumé .............................................................................................................. v Chapter 2: Literature review and Objectives 2.1. Literature review ..................................................................................................... 3 2.1.1. Introduction .................................................................................................... 3 2.1.2. Effect of main cereal of the diets on productive performance ....................... 4 2.1.3. Effect of feed form on productive performance ............................................. 6 2.1.4. Effect of cereal type of the diets on digestive tract traits .............................. 8 2.1.5. Effect of feed form on digestive tract traits .................................................. 9 2.2. Objectives ................................................................................................................ 11 2.3. Reference ................................................................................................................. 12 Chapter 3: Influence of the main cereal and feed form of the diet on performance and digestive tract traits of brown-egg laying pullets from hatching to 17 week of age. 3.1. INTRODUCTION .................................................................................................. 19 3.2. MATERIALS AND METHODS ........................................................................... 21 3.2.1. Husbandry, Diets, Feeding Program, and Experiment Design .................... 21 3.2.2. Growth Performance .................................................................................... 22 3.2.3. Gastrointestinal tract traits ........................................................................... 22 3.2.4. Laboratory analysis ...................................................................................... 23 3.2.5. Statistical analysis ........................................................................................ 23 3.3. RESULTS ................................................................................................................ 24 3.3.1. Growth Performance .................................................................................... 24 3.3.2. Gastrointestinal Tract traits .......................................................................... 25 3.3.3. Body and tarsus Measurements .................................................................... 26 3.4. DISCUSSION .......................................................................................................... 27 3.4.1. Growth Performance .................................................................................... 27 3.4.2. Gastrointestinal tract traits .......................................................................... 30 3.4.3. Body and tarsus Measurements .................................................................... 32 3.5.CONCLUSON .......................................................................................................... 32 3.6. Reference ................................................................................................................. 33 Chapter 1: Abstract ii Chapter 1: ABSTRACT ABSTRACT The influence of the main cereal and feed form of the diet on performance and gastrointestinal tract (GIT) traits was studied in brown-egg laying pullets from hatching to 17 wk of age. There were 8 treatments arranged as a 2×4 factorial with 2 main cereals (corn vs. wheat) and 4 feeding programs that consisted in feeding crumbles to pullets from 1 to 5 wk, 1 to 10 wk followed by mash to 17 wk of age, and feeding crumble or mash continuously from 0 to 17 wk of age. Each treatment was replicated 9 times (17 pullets per replicate). From wk 0 to 5 pullets fed wheat had higher BWG (P < 0.001) and better FCR (P < 0.001) than pullet fed corn but the differences disappeared with age. Pullets fed crumbles continuously showed higher BW gain (14.0 vs.13.3 g/d; P < 0.001) and better FCR (4.28 vs. 4.44; P < 0.001) than pullets feed mash continuously, with pullets fed the other 2 treatments being intermediate. When feed form was changed from mash to crumble at any age, pullets performance was improved. Pullets fed corn had heavier GIT and gizzard than pullets fed wheat (P < 0.001) but body length, tarsus length and length of the small intestine were not affected. Pullets fed mash from wk 0 to 17 were longer (P < 0.001) and higher (P < 0.001) relative weight (% BW) of the gizzard and of the GIT but lower (P < 0.001) gizzard pH than pullets fed crumbles continuously, with pullets fed the others 2 treatments being intermediate. We conclude that wheat and corn can be used indistinctly in diets for pullets. Also, feeding crumbles improved growth performance of pullets but reduced the relative weight of the GIT and of the gizzard. The GIT of the pullets adapts quickly to changes in feed form. Feeding crumble improves growth performance without any negative effects on pullet uniformity. iii Chapter 1: Resumen Resumen El objetivo general de esta Tesis de Máster fue estudiar la influencia del tipo de cereal y la forma de presentación del pienso sobre los parámetros productivos y al desarrollo del tracto digestivo en pollitas de 0 a 17 semanas (sem) de vida. El diseño experimental fue complemente al azar con 8 tratamientos organizados de forma factorial con 2 cereales base (trigo vs. maíz) y 4 programas de alimentación, que consistieron en modificar la forma de presentación del pienso (miga vs. harina) según la fase de recría (1-5 sem, 510 sem y 10-17 sem de edad). Dos de los tratamientos consistieron en suministrar los piensos durante toda la recría (0 -17 sem) bien en forma de harina o bien en forma de migas. Los otros dos tratamientos consistieron en suministrar el migas de 0 a 5sem o de 0 a 10 sem seguido del suministrar en harina hasta las 17 sem. Se utilizaron 9 réplicas por tratamiento y la unidad experimental fue la jaula con 17 pollitas. De 0 a 5 sem, las pollitas alimentadas con trigo tuvieron una ganancia media diaria (GMD) mayor (P < 0.001) y un índice de conversión (IC) mejor (P < 0.001) que las pollitas alimentados con maíz. En el global de la prueba (0 a 17 sem), el tipo de cereal no afectó de forma significativa a ninguno de los parámetros estudiados. La presentación del pienso influyó sobre los parámetros productivos a lo largo del periodo experimental. De hecho, el cambio del pienso de miga a harina se provocó una pérdida de rendimientos productivos en el periodo posterior. De 0 a 17 sem de vida, la GMD fue mayor (14.0 vs.13.3 g/d; P < 0.001) y el IC fue mejor (4.28 vs. 4.44; P < 0.001) para las pollitas que consumieron migas durante toda la prueba que para las que consumieron harina con las pollitas que recibieron los 2 tratamientos mostraron resultados intermediaros. El desarrollo del GIT y de la molleja fue mayor con las pollitas alimentadas con piensos basados en maíz que las pollitas alimentadas con piensos basados en trigo. La alimentación en miga redujo el peso relativo del tracto gastrointestinal y de la molleja (P < 0.001). El pH del contenido de la molleja fue inferior en las pollitas que recibieron harina durante todo el periodo de recría (P < 0.01). A 17 sem de edad La longitud del cuerpo y del tarso fue mayor con las pollitas alimentadas con piensos en forma de harina que para las que consumieron piensos en migas de continua. Por otro lado, las pollitas que recibieron los 2 tratamientos combinados mostraron resultados intermediaros. iv Chapter 1: Resumen En base a los resultados obtenidos, al menos de 40% el trigo suplementado con enzimas puede utilizarse en sustitución de maíz en piensos de pollitas de 0 a 17 sem de edad. Por otro lado, la utilización de piensos en migas mejora la productividad de las pollitas pero podría afectar negativamente el desarrollo del aparato digestivo. Las pollitas adaptan rápidamente su sistema digestivo, consumo de pienso y productividad general, a cambios en la presentación del pienso. v Chapter 1: Résumé Résumé L’objectif général de cette Thèse de Master était d'étudier l'influence de type de céréale, et la forme de présentation de l aliment qui pourraient affecter les performances productives et le développement du tube digestif des poulettes brunes pour la production d’œufs commerciaux., 8 traitement organisés de forme factorielle avec 2 types de céréales (maïs contre blé) et 4 programmes de alimentation qui consistent a modifier la forme de présentation de l aliment (farine contre miette) durant les 3 périodes de l élevage (0-5 semaines, 5-10 semaines et 10-17 semaines). Deux de 4 traitements consistent de distribuer l’aliment sous forme de miette ou sous forme de farine durant les trois périodes d’élevage. Les deux autres traitements consistent de combiner la forme de présentation de l’aliment tout le long de la période de l'essai. Un de deux traitements consiste á distribuer l’aliment sous forme de miette seulement durant la première période (0-5 semaine) et l’autre consiste á offrir l’aliment sous forme de miette durant les deux périodes de l`élevage (0-5 semaines, 5-10 semaines) et après tous les régimes ont été offerts sous la forme farineuse et la seule différence entre les régimes était la céréale de base utilisée. Chaque traitement a été répété 6 fois (17 poulettes par répétition). De 0 à 5 semaines, les poulettes alimentées avec des régimes à base de blé ont eu un gain de poids plus élevé (14.0 vs.13.3 g/d; P < 0.001) et un indice de conversion meilleur (P < 0.001) à ceux des poulettes alimentées avec des régimes à base de maïs. Durant la période global le type de céréale n’affecte pas les paramètres productifs. La forme de présentation de l’aliment affecte les paramètres productifs chez les poulettes durant les différentes périodes de l’élevage. Les poulettes alimentées à base de miette durant tout le long de la période de l élevage et ont eu un gain de poids plus élevé (14.0 vs.13.3 g/d; P < 0.001) et un indice de conversion meilleur que les poulettes alimentées à base de farine (4.28 vs. 4.44; P < 0.001). On détecte une réduction des paramètres productifs lorsqu’on change la forme de présentation d’aliment (de miette à farine). Le poids relatif (% BW) du gésier et du tube digestif étaient plus élevés (P < 0.001) chez les poulettes alimentées avec des régimes à base de maïs que chez les poulettes alimentées avec des régimes à base de blé. Durant la période global, L’utilisation de l’aliment farineux augmente le poids relatif du tube digestif et du gésier (P < 0.001) aussi bien que Le pH du contenu de gésier à 17 semaines d’âge n'a vi Chapter 1: Résumé pas été affecté par la céréale de base utilisée mais il était inférieur chez les poulettes alimentées à base de farine. Les poulettes alimentées avec le traitement combiné montrant des résultats intermédiaire. Nous concluons que 40% de blé peut substituer le maïs dans les régimes de poulette avec une légère réduction du gain de poids vif. Aussi, l’utilisation de l'aliment granulé depuis 0 jusqu’à 17 semaines d'âge a augmenté le gain de poids vif et le pH du gésier. La présentation de l’aliment granulé a réduit le poids relatif du gésier et la longueur du tube digestif à 17 semaines d'âge. Chapter 2: Literature review and Objectives 3 Chapter 2: Literature review and Objectives 2.1. Literature review and objectives 2.1.1. Introduction The egg industry changes and evolves quickly due to increased demand for quality, technology changes, and pressures from consumers and government regulators. Also, the egg industry continues to grow as the egg offers consumers a source of protein at low cost. Egg production and consumption continues to grow in most countries around the world as the international trade of eggs is relatively insignificant because of handling difficulties with some exceptions mainly in the European Union. The demand for processed egg products continues will probably to increase (FAO Stat, 2011). Spain is one of the most important egg producers in Europe with more than 35 millions of industrial laying hens, and more than 883 thousand tones of egg produced per year (FAO Stat, 2011). Egg size is especially important in Spain because consumers show a clear preference for large eggs (Grobas et al., 1999). The profitability of the egg industry depends mostly of 3 factors; number of eggs per hen housed, size of the eggs produced, and percentage of eggs that reach the table of the final consumer. To improve egg rate and the quality of the eggs, one of the most critical points is the management and the nutrition of the pullets during the rearing phase. Pullets should reach sexual maturity with a BW and uniformity as recommended by the genetic companies that market them. In fact, the rate of egg production and the percentage of large eggs are positively related to feed intake and the BW of pullets during the rearing period. Summers and Lesson (1983) found that heavier pullets laid significantly more and heavier eggs than did lighter pullets at the beginning of the egg production cycle. Age of sexual maturity of pullets can be advanced considerably by breeding programs, light stimulation, or nutrition, and by a combination of the three. Decreasing the age of sexual maturity increases the number of eggs laid with the potential disadvantage of the production of a greater proportion of small eggs. Changing feeding practices, including the choice of raw materials and the form, seems to be promising ways to reduce cost of production. Indeed, the feed represents the greatest part of the economic cost of eggs production (Boggia et al., 2010). 10 Chapter 2: Literature review and Objectives Jensen and Becker (1965) suggested that the pelleting process to some extent gelatinized starch. Also, pelleting of diets can reduce their particle size substantially (Engberg et al., 2002). The combination of particle size reduction and starch gelatinization may expose feed particles more efficiently to enzymatic digestion, which may explain the improved AME and the apparent crude protein (CP). Mateos et al. (2002) and González-Alvarado et al. (2008) suggested that a more functional gizzard might result in more reflux and better mixing of the digesta and endogenous enzymes in the GIT. Therefore, a well-developed gizzard might increase nutrient digestibility and help to maintain a healthy microbiota population in the GIT (Gabriel et al., 2008; Santos et al., 2008). Choi et al. (1986) and Nir et al. (1994a, b) observed that feeding crumbles or pellets reduced the relative weight (g/kg of BW) of the gizzard compared with feeding mash. Nir et al. (1994b) observed that feeding crumbles or pellets to broilers reduced gizzard weight with respect to feeding mash. In pullets, Frikha et al. (2009b) observed that feeding pellets to pullets reduced the RW of the gizzard, proventriculus and digestive tract at 45 d of age. Also, the relative length (RL) of the small intestine (SI), jejunum, ileum and ceca was reduced at this age. However, at 120 d of age, the only differences observed were for the RW of gizzard and proventriculus that were heavier for pullets previously fed mash than for those fed pellets. Gizzard pH at 120 d of age was not affected by diet. Frikha et al. (2009a), the digestive tract and the gizzard were heavier in pullets fed mash at 45 d of age than in pullets fed pellets. In addition, the small intestine and the ceca were longer in pullets fed mash than in those fed pellets. The information available comparing effect of changing feed form on productive performance and digestive tract traits development (GIT), in broilers is abundant but scarce in pullets. 11 Chapter 2: Literature review and Objectives 2.2. Objectives The present Master Thesis has been carried in collaboration between department of animal production (UPM) and Cantos it is the largest egg producing company of Spain with more than 5 millions hens. 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Res.8:16-24. 15 Chapter 2: Literature review and Objectives Peron, A., J. Gomez, S. Mignon-Grasteau, N. Sellier, J. Besnard, M. Derouet, H. Juin, and B. Carre. 2006. Effects of wheat quality on digestion differ between the D+ and D− chicken lines selected for divergent digestion capacity. Poult. Sci. 85:462-469. Pérez-Bonilla , M. Frikha , S. Mirzaie , J. Garcia , and G. G. Mateos. 2011. Effects of the main cereal and type of fat of the diet on productive performance and egg quality of brown-egg laying hens from 22 to 54 weeks of age. Poult. Sci. 90:2801-2810. Pettersson, D., H. Graham, and P. Aman. 1991. The nutritive value for broiler chickens of pelleting and enzyme supplementation of a diet containing barley, wheat and rye. Anim. Feed Sci. Technol. 33:1-14. Pirgozliev, V. R., C. L. Birch, S. P. Rose, P. S. Kettlewell, and M. R. Bedford. 2003. Chemical composition and the nutritive quality of different wheat cultivars for broiler chickens. Br. Poult. Sci. 44:464-475. Quentin, M., I. Bouvarel, and M. Picard. 2004. Shortand long-term effects of feed form on fastand slow-growing broilers. J. Appl. Poult. Res. 13:540-548. Rama Rao, S. V., M. R. Reddy, N. K. Prarharaj, and G. Shyam Sunder. 2000. Laying performance of broiler breeder chickens fed various millets or broken rice as source of energy at a constant nutrient intake. Trop. Anim. Health Prod. 32:329338. Reece, F. N., B. D. Lott, and J. W. Deaton. 1985. The effect of feed form, grinding method, energy level, and gender on broiler performance in a moderate (21ºC) environment. Poult. Sci. 64:1834-1839. Rougiere, N., and B. Carre. 2010. Comparison of gastrointestinal transit times between chickens from D+ and D − genetic lines selected for divergent digestion efficiency. Animal 4:1861-1872. Ruiz, N., J. E. Marion, R. D. Miles, and R. B. Barnett. 1987. Nutritive value of new cultivars of triticale and wheat for broiler chick diets. Poult. Sci. 66:90-97. Safaa, H. M., E. Jiménez-Moreno, D. G. Valencia, M. Frikha, M. P. Serrano, and G. G. Mateos. 2009. Effect of main cereal of the diet and particle size of the cereal on productive performance and egg quality of brown egg-laying hens in early phase of production. Poult. Sci. 88:608-614. Salih, M. E., H. L. Classen, and G. L. Campbell. 1991. Response of chicken fed on hullless barley dietary β-glucanase at different ages. Anim. Feed Sci. Technol. 33:139-149. Santos, F. B. O., B. W. Sheldon, A. A. Santos Jr., and P. R. Ferket. 2008. Influence of housing system, grain type, and particle size on Salmonella colonization and shedding of broilers fed triticale or corn-soybean meal diets. Poult. Sci. 87:405420. Serrano, M. P., D. G. Valencia, J. Méndez, and G. G. Mateos. 2012. Influence of feed form and source of soybean meal of the diet on growth performance of broilers from 1 to 42 days of age. 1. Floor pen study. Poult. Sci. 91:2838-2844. Serrano, M. P., M. Frikha, J. Corchero, and G. G. Mateos. 2013. Influence of feed form and source of soybean meal on growth performance, nutrient retention, and digestive organ size of broilers. 2. Battery study. Poult. Sci. 92:693-708. Soler, J. J., and M. Soler. 1993. Diet of the red-billed chough Pyrrhocorax pyrrhocorax in South-East Spain. Bird Study 40:216-222. 16 Chapter 2: Literature review and Objectives Summers, J. D., and S. Leeson, 1983. Factors influencing early egg size. Poultry Sci. 62:1155-1159. Summers, J. D., and S. Leeson. 1994. Laying hen performance as influenced by protein intake to sixteen weeks of age and body weight at point of lay. Poult. Sci.73:495-501. Svihus, B., K. H. Klovstad, V. Perez, O. Zimonja, S. Sahlstrom, and R. B. Schuller. 2004. Physical and nutritional effects of pelleting of broiler chicken diets madefrom wheat ground to different coarseness by the roller mill and hammer mill.Anim. Feed Sci. Technol. 117:281-293. Svihus, B. 2011. The gizzard: Function, influence of diet structure and effects on nutrient availability. World´s Poult. Sci. J.67:207-224. Wilson, k.j, R.S. Beyer, J.R. Froetschner and K.C, Behnke. 2001. Effect of feed conditioning method on broiler performance. Ph.D. Thesis, Kansas State University, Manhattan, K.S. Zang, J.J., Piao, X.S., Huang, D.S., Wang, J.J., Ma, X., Ma, Y.X. 2009. Effects of feed particle size and feed form on growth performance, nutrient metabolizability and intestinal morphology in broiler chickens. Asian Australas. J. Anim. Sci. 22, 107-112. Chapter 3: Influence of the main cereal and feed form of the diet on performance and digestive traits of brown-egg laying pullets from hatching to 17 wk 19 Chapter 3: Main Cereal and Feed Form for Pullets 3.1. INTRODUCTION Corn and wheat are the most common used cereals in poultry diets. Corn has less protein (7.7 vs. 11.2%) but more energy (3,260 vs. 3,150 kcal AMEn /kg) than wheat (Fundación Española Desarrollo Nutrición Animal, 2010). However, chemical composition and energy availability is more variable for wheat than for corn (Kim et al., 1976; Mollah et al., 1983; Gutierrez-Alamo et al., 2008) which reduces the interest of wheat as an ingredient in poultry feeding. Several reports have compared diets based on corn or wheat on productive performance of broilers (Crouch et al., 1997; Mathlouthi et al., 2002) and laying hens (Lazáro et al., 2003a; Pérez-Bonilla et al., 2011). In most cases, authors concluded that wheat can be used in substitution of corn without any negative effects on performance, provided that the diet is supplemented with enzymes. However, Frikha et al. (2009b) reported that pullets fed wheat diets from hatching to 120 d of age had lower BWG than pullets fed corn diets, although no differences were observed for ADFI or FCR. Also, they observed that at 6 wk of age the gizzard was heavier in 6 wk-old pullets fed corn than in pullets fed wheat. The authors had not found any other report on the effects of the main cereal of the diet on performance and gastrointestinal tract (GIT) traits of brown-egg laying pullets from hatching to 17 wk of age fed mash or crumbles diets. Under commercial conditions, broilers are frequently fed crumbles from 1 to 3 wk and then pellets to slaughter, a practice that results in improved BW gain (BWG) and FCR (Amerah et al., 2007; Cerrate et al., 2008; Serrano et al., 2012, 2013). However, the information available on the effects of feed form on pullet performance is scarce. Frikha et al. (2009b) reported that from hatching to 120 d of age, BWG and ADFI were higher in pullets that were fed pellets from 1 to 45 d of age than for pullets that were fed mash. In addition, from 46 to 85 d of age pullets that were fed pellets previously showed higher BWG than pullets that were fed mash. 26 Chapter 3: Main Cereal and Feed Form for Pullets At 5 wk and 10 wk of age, pullets fed mash had heavier GIT and gizzard (P < 0.01) than pullets fed crumbles (Table 5). An increase in the relative weight of the GIT and gizzard was observed at all times in which pullets were changed from crumble to mash feeds (Tables 5 and 6, respectively). Gizzard digesta content was higher and gizzard pH was lower (P < 0.01) in all of the 3 feeding periods considered in pullets fed mash than in pullets fed crumbles continuously (Table 6). At 5 and 10 wk of age, pullets fed mash continuously had heavier (P < 0.01) gizzards than pullets fed crumbles continuously. The digest content of the gizzard was increased (P < 0.01) and gizzard pH (P < 0.01) was reduced in pullets fed mash as compared with pullets fed crumbles. At 17 wk of age, neither the SI or the ceca length were affected by feed form but the jejunum was shorter (P < 0.05) in pullets fed crumbles continuously than in pullets fed mash continuously, with pullets from the others treatment being intermediate (Table 7). At 5 wk of age the SI was shorter at 5 wk (P = 0.09) and at 10 wk (P < 0.001) of age in pullets fed crumbles than in pullets fed mash continuously with most of the differences observed for the ileum and jejunum length. 3.3.3. Body and Tarsus Measurements No interaction between main cereal and feed form of the diet was detected for any measurement for body and tarsus and therefore, only main effects are presented (Table 8). Main cereal: The main cereal of the diet did not affect the relative length of the pullets or of the tarsus at 5 or 10 wk of age. However, at 17 wk of age pullets fed corn tended to be longer (48.9 vs. 48.1 cm/kg BW; P < 0.01) than pullets fed wheat. Feed form: At 17 wk of age, pullets fed mash continuously were longer and had longer tarsus than pullets fed crumbles continuously (P < 0.05) with most of the differences observed already at 5 wk of age. At this age, Pullets fed mash were longer 27 Chapter 3: Main Cereal and Feed Form for Pullets (P < 0.01) and had longer (P < 0.01) tarsus than pullets fed crumbles. At 10 wk of age, pullets fed mash had longer tarsus (P < 0.01) than pullets fed crumbles, but no differences in pullet length were detected. 3.4. DISCUSSION The gross energy, crude protein, and ash content of the experimental diets within each feeding period were similar to calculated values, indicating that the ingredients were mixed correctly. The GMD of the diets, within each feeding period, was similar for the corn and the wheat based diets and was lower from 0 to 5 wk of age than from 5 to 10 wk or 10 to 17 wk, consistent with the lower screen used to grind the cereals of the starter feeds (Table 2). 3.4.1. Growth Performance Main cereal: For the entire experimental period, the main cereal of the diet did not affect productive performance of the pullets. Therefore wheat supplemented with enzymes can be used as a substitute of corn in diets for pullets. These results agree with data from studies conducted in broilers (Ruiz et al., 1987; Mathlouthi et al., 2002), pullets (Frikha et al., 2010), and laying hens (Ciftci et al., 2003; Lázaro et al., 2003a; Safaa et al., 2009; Pérez Bonilla et al., 2011). The information provided confirms that wheat composition is more variable than that of corn and therefore, poultry might respond differently to different wheat varieties. For example, Crouch et al. (1997) compared corn and two varieties of wheat at 40% of inclusion in mash diets for broilers and found that BWG and FCR were hindered with one of the two wheats but not with the other. Pullets fed wheat supplemented with enzymes had higher BWG at 5 wk of age and had better FCR from wk 0 to 10 of age than pullets fed corn consistent with data of Moran et al. (1993) who observed better growth performance with wheat than with corn 28 Chapter 3: Main Cereal and Feed Form for Pullets from 1 to 42 d of age in broilers. These authors observed that the inclusion of adequate enzymes improved more performance of broilers that were fed low-quality wheat than of those fed the high quality traits. Frikha et al. (2009b) compared wheat and corn as main cereal of the diets in pullets from 1 to 17 wk of age. In the first experiment, the authors reported better performance at 17 wk of age with corn whereas no differences were detected in the second experiment. Ruiz et al. (1987) reported similar BWG and feed conversion ratio in broilers fed mash when corn was substituted by wheat. The reason for the differences in BWG for pullets fed wheat than for pullets fed corn among experiment is unknown. Wheat contains a high and variable amount of nonstarch polysaccharides (NSP), which are known to increase digesta viscosity and reduce productive performance in poultry (Làzaro et al., 2003a, b; Garcia et al., 2008). Làzaro et al. (2003a, 2004) reported that enzymes reduced digesta viscosity and improve nutrient digestibility and feed intake in laying hens and broilers fed high NSP cereals. In laying hens, most reports indicate that wheat conveniently supplemented with enzymes can be used successfully as the main ingredient of the diet (Liebert et al., 2005; Safaa et al., 2009; Perez-Bonilla et al., 2011). From 10 to 17 wk of age, pullets fed corn had higher BWG (P < 0.01) than pullets fed wheat but no differences were observed for ADFI or FCR, data that agree with results of Frikha et al. (2009b) in pullets from 1 to 120 d of age. Ouart et al. (1986) indicated that ADFI and FCR of SCWL hens from 68 to 71 wk of age were not affected when 37% corn was substituted by wheat. Pérez-Bonilla et al. (2011) observed that ADFI and FCR of brown-egg laying hens from 22 to 54 wk of age were similar when fed corn than when fed wheat based diets. 29 Chapter 3: Main Cereal and Feed Form for Pullets Similarly, Mathlouthi et al. (2002) reported same performance when 60 % corn of a broiler diet was substituted by a mixture of 40 % wheat and 20 % barley supplemented with enzymes. In contrast, Kim et al. (1976) found that SCWL hens fed a diet based on corn consumed more feed and produced heavier eggs than hens fed diet based on wheat from 21 to 43 wk of age, although BW was not affected. Feed form: For the entire experiment, Pullets fed crumbles continuously had higher BWG and better FCR than pullets feed mash continuously, with pullets changed from crumble to mash at 5 or 10 wk of age being intermediate. The effects of feeding crumbles or pellets to broilers on growth performance have been studied in detail (Cerrate et al., 2008, 2009; Serrano et al., 2012, 2013). Crumbling increases feed density and improves the texture of the feed which might increase ADFI in broilers. In addition, because of its small particle size pelleting facilitates the contact and access of endogenous enzymes to nutrients. In this addition, Gracia et et al. (2003) reported that heat processing modifies starch, protein, and fiber structure of the cereal and improves accessibility of enzymes to nutrients facilitating its digestibility. However, the mild temperatures applied during the pelleting process might not affect at any high extent nutrient digestibility. Abdollahi et al. (2011) observed that from 4 to 21 d of age BWG was 10.3% higher in broilers fed pellets than in broilers fed mash. In pullets, Gous and Morris (2011) reported that pullets fed crumbles from 1 to 4 wk and then pellets from 5 to 20 wk of age consumed 2% less feed and were 6% heavier at 20 wk than pullets fed mash. In contrast, Hamilton and Proudfoot (1995) found that BW at 20 wk of age was higher for Leghorn hens fed mash diets as compared with hens fed crumbled diets. For the entire experiment period, pullets that were fed crumbles from 1 to 5 wk of age had lower ADFI than pullets that were fed crumbles or mash continuously with pullets fed crumbles from 1 to 10 wk of age being intermediate. 30 Chapter 3: Main Cereal and Feed Form for Pullets In fact, ADFI was reduced at any time pullets were changed from crumbles to mash feed. The reason for the lower ADFI any time that pullets were changed from crumbles to mash feed is not known. Feed wastage was not measured in this research but probably it was higher for mash than for crumbles as has been reported in piglets (Medel et al., 2004; Berrocoso et al., 2013) and broilers (Serrano et al., 2013). Serrano et al. (2012) reported that from 21 to 42 d of age, broilers fed pellets that were fed mash previously, had higher ADFI than broilers that were fed crumbles or pellets. Body weight uniformity was not affected by the main cereal or feed form at any age in the rearing phase, results agree with Frikha et al. (2009b) who did not observed any effect on BW uniformity at any age in pullets fed corn or wheat in pellet or mash form. 3.4.2. Gastrointestinal Tract Traits Main cereal: At 17 wk of age, GIT and gizzard were heavier for pullets fed corn than for pullet fed wheat. At 5 wk of age, pullets fed corn had heavier gizzards than those fed wheat, results that agree with data of Amerah et al. (2008) in broilers. Also, pullet fed corn had higher gizzard digest contents than pullets fed wheat, consistent with the GMD of the 2 diets. The higher weight of the gizzard in pullets fed corn might be related to the GMD of the diets as well as differences in the structure of the endosperm that is harder and more difficult to grind for corn than for wheat (Dombrink-Kurtzman and Bietz, 1993; Dobraszczyk et al., 2002). The percentage of fine particles (< 160 μm) was higher for the wheat than for the corn diets. Fine particles passes faster through the gizzard than coarse particles (Hetland et al., 2002; Svihus and Hetland, 2002) and consequently, gizzard contents should be reduced when wheat replaces corn. Feed form: In the current experiment, the relative weight of the GIT and the gizzard increased when pullets were changed from crumbles to mash feeds. The data indicate that crumbling might have a negative effect on the development of the GIT of pullets whether they are fed corn or wheat. 31 Chapter 3: Main Cereal and Feed Form for Pullets Also, the relative digest content of the gizzard was higher when the diets were fed in mash than when feed in crumble form, results that agree with data of Frikha et al. (2009b). Nir et al. (1994) observed that feeding crumbles or pellets to broilers reduced the relative weight of the gizzard as compared with feeding mash. The GMD of the feed is smaller with crumbles than with mash which results in faster transit time throughout the GIT. Similar results have been published by Jiménez-Moreno et al. (2008) in broilers. On the other hand, gizzard pH was lower for pullets fed mash than for pullets fed crumbles, results that agree with data of Frikha et al. (2009b) in pullets fed similar type of diets. In contrast, Dahlke et al. (2003) in broilers did not observe any effect on gizzard pH when pelleted diets based on corn, varying in GMD from 340 μm to 1,120 μm, were used. In the current research, the changes in feed form from crumbles to mash at 5 and 10 wk of age might have reduced the negative effects of pelleting on gizzard pH. At 10 wk of age, feeding mash increased the relative length of all the segments of the small intestine, except that of the ileum and the ceca, results that agree in part with data of Frikha et al. (2009a) who reported that feeding pellets from 1 to 45 d of age reduced the RL of the jejunum and ileum in Hy-Line Brown pullets. However, at 17 wk of age the only differences observed was for the RL of the jejunum that was higher for pullets fed mash continuously than in pullets fed crumbles continuously, with pullets changed from crumble to mash at 5 or 10 wk of age being intermediate, results that agree with data of Nir et al. (1995) found that pelleting reduced by 15% the RL of the jejunum and ileum of broilers. Similar results have been reported by Amerah et al. (2007). 32 Chapter 3: Main Cereal and Feed Form for Pullets 3.4.3. Body and Tarsus Measurement In the current experiment, The main cereal of the diet did not affect the relative length of the pullets and of the tarsus but pullets fed mash continuously were longer and had longer tarsus than pullet mash continuously results that agree with data of Amerah et al. (2007) reported that the improvement in broiler performance observed with pelleting was associated with a decrease in the RL of the GIT. To our knowledge, this is the first report indicating that feeding crumbles to pullets from hatching to 17 wk of age reduced the relative length of the pullets, the tarsus and the weight of the GIT but increased gizzard pH. 3.5. CONCLUSION We conclude that, the main cereal of the diet did not affect pullet performance from 0 to 17 wk of age. 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Influence of main cereal and feed form of the diet on the relative weight (% BW) of the gastrointestinal tract (GIT) of the pullets Cereal Feed form1 5 week 10 week 17 week 0-5 week 5-10 week 10-17 week BW2 GIT3 BW GIT BW GIT Corn C C C 335 20.6 967 13.7 1,339 11.4 C C M 348 20.8 970 13.7 1,314 12.1 C M M 329 21.0 891 15.1 1,314 12.4 M M M 301 22.4 888 15.9 1,290 13.0 Wheat C C C 339 20.3 968 14.2 1,409 10.5 C C M 338 20.2 969 14.2 1,328 11.6 C M M 335 21.1 926 15.4 1,301 12.2 M M M 325 21.9 926 15.5 1,318 12.5 Main effect Cereal Corn 329 21.2 929 14.6 1,314 12.2a Wheat 335 20.8 947 14.8 1,339 11.7b Feed form C C C 337a 20.4b 969a 13.9b 1,374a 11.0c C C M 344a 20.5b 970a 14.0b 1,321ab 11.9b C M M 333ab 21.0ab 909b 15.2a 1,307b 12.3ab M M M 314ab 22.2a 907b 15.7a 1,304b 12.7a SEM4 22.43 1.45 54.01 0.81 61.35 0.66 Probability Cereal 0.252 0.317 0.171 0.256 0.092 0.002 Feed form 0.001 0.002 0.001 0.001 0.003 0.001 cereal *Feed form 0.174 0.912 0.573 0.259 0.241 0.462 a-b-c Within a column, means without a common superscript differ (P < 0.05). 1C= crumble; M= mash. 2Body weight. 3 Includes the weights of the digestive tract (from the beginning of the proventriculus to cloaca, with digesta content), the liver, and the pancreas. 4SEM (9 replicates of 2 pullets each per treatment). 43 Table 6. Influence of main cereal and feed form of the diets on the relative weight (% BW) of the gizzard, gizzard digest content (% gizzard weight) and gizzard pH of the pullets Cereal Feed form1 5 week 10 week 17 week 0-5 week 510 week 1017 week Weight (% BW) Gizzard content pH Weight (% BW) Gizzard content pH Weight (% BW) Gizzard content pH Corn C C C 4.6 31.3 3.42 2.7 16.3 3.96 3.2 19.6 3.63 C C M 4.5 29.8 3.38 2.5 15.0 3.73 4.1 28.4 3.33 C M M 4.6 28.1 3.51 4.9 31.9 2.77 4.4 28.7 3.06 M M M 5.9 34.3 3.01 5.0 32.6 2.61 4.6 28.5 3.21 Wheat C C C 3.9 27.1 3.44 3.0 21.3 3.71 2.5 15.0 4.11 C C M 4.1 27.5 3.33 3.0 18.8 3.68 3.7 27.1 3.20 C M M 4.5 28.6 3.26 4.7 30.3 2.65 4.2 29.3 3.16 M M M 5.7 34.1 2.83 4.9 30.2 2.75 4.2 26.6 3.14 Main effect Cereal Corn 4.9a 30.9 3.30 3.8 24.0 3.3 4.1a 26.3a 3.31 Wheat 4.6b 29.3 3.20 3.9 25.2 3.2 3.7b 24.5b 3.40 Feed form C C C 4.3b 29.2b 3.40a 2.9b 18.8b 3.84a 2.8c 17.3b 3.87a C C M 4.3b 28.7b 3.30a 2.8b 16.9b 3.70a 3.9b 27.8a 3.26b C M M 4.6b 28.4b 3.40a 4.8a 31.1a 2.71b 4.2a 29.0a 3.11b M M M 5.8a 34.1a 2.90b 5.0a 31.4a 2.68b 4.4a 27.5a 3.17b SEM2 0.53 5.09 0.35 0.457 4.902 0.38 0.57 3.56 0.46 Probability Cereal 0.007 0.196 0.215 0.280 0.256 0.467 0.001 0.034 0.389 Feed form 0.001 0.003 0.001 0.001 0.001 0.001 0.001 0.001 0.001 cereal *Feed form 0.395 0.506 0.676 0.052 0.259 0.483 0.163 0.183 0.211 a-b-c Within a column, means without a common superscript differ (P < 0.05). 1C= crumble; M= mash. 2SEM (9 replicates of 2 pullets each per treatment). 44 Table 7. Influence of main cereal and Feed form of the diets on the relative length (cm/ kg BW) of the organs of the SI and ceca of pullets 1C=crumble; M=mash. 2Duodenum. 3Jejunum. 4Small intestine. 5 SEM (9 replicates 2 pullets each per treatment). Feed form1 5 week 10 week 17 week Cereal 0-5week 5-10week 10-17week Duod2 Jejun3 Ileum SI4 Ceca Duod Jejun Ileum SI Ceca Duod Jejun Ileum SI Ceca Corn C C C 30.9 158.6 128.0 317.5 33.0 10.8 61.7 48.5 121.0 13.4 7.9 45.5 36.9 90.3 12.9 C C M 28.5 152.6 124.7 305.8 32.1 10.9 61.8 50.5 123.3 13.5 7.7 45.7 37.2 90.7 12.3 C M M 30.0 160.9 132.4 323.2 33.0 11.8 66.1 50.4 128.4 13.7 8.0 47.4 38.1 93.5 12.3 M M M 33.0 171.0 142.7 346.7 35.5 11.8 67.7 53.3 132.8 14.1 7.7 48.7 39.5 95.9 13.2 Wheat C C C 30.7 155.3 127.2 313.2 32.8 10.5 61.4 49.6 121.5 13.0 7.2 44.0 36.4 87.6 12.3 C C M 29.9 157.3 128.9 316.1 32.7 10.9 62.3 49.9 123.1 13.2 7.8 48.1 37.7 93.7 12.9 C M M 30.0 157.4 127.1 314.5 33.1 11.6 65.1 50.2 126.9 14.1 7.8 47.6 38.4 93.9 13.2 M M M 31.5 165.4 128.3 325.2 33.7 11.2 64.2 48.1 123.5 13.6 7.7 48.6 38.1 94.4 12.9 Main effects Cereal Corn 30.6 160.8 132.0 323.3 33.4 11.3 64.3 50.7 126.4 13.7 7.8 46.8 37.9 92.6 12.7 Wheat 30.5 158.8 127.9 317.3 33.1 11.1 63.3 49.4 123.7 13.5 7.6 47.1 37.7 92.4 12.8 Feed form C C C 30.8ab 157.0 127.6 315.4 32.9 10.7b 61.5b 49.0 121.3b 13.2b 7.5 44.7b 36.7 88.9 12.6 C C M 29.2b 154.9 126.8 311.0 32.4 10.9b 62.1b 50.2 123.2b 13.4ab 7.8 46.9ab 37.5 92.2 12.6 C M M .30.0ab 159.1 129.8 318.9 33.0 11.7a 65.6a 50.3 127.6a 13.9a 7.9 47.5ab 38.2 93.7 12.7 M M M 32.3a 168.2 135.5 335.9 34.6 11.5a 66.0a 50.7 128.1a 13.9a 7.7 48.7a 38.8 95.2 13.7 SEM5 3.0 15.63 13.88 31.17 3.21 0.98 4.82 5.16 9.58 1.09 0.88 4.03 4.1 8.14 1.04 Probability Cereal 0.944 0.610 0.217 0.415 0.711 0.088 0.183 0.538 0.105 0.254 0.282 0.769 0.773 0.906 0.648 Feed form 0.025 0.07 0.241 0.095 0.212 0.001 <0.001 0.498 0.005 0.015 0.625 0.037 0.434 0.134 0.498 Cereal* Feed form 0.584 0.774 0.232 0.501 0.677 0.706 0.348 0.645 0.120 0.365 0.516 0.555 0.891 0.743 0.129 45 Table 8. Influence of main cereal and feed form of the diets on the relative length (cm/ kg BW) of the pullets and the tarsus. Feed form1 5 week 10 week 17 week Cereal 0-5 week 5-10 week 10-17 week Pullet2 length Tarsus3 length Pullet length Tarsus length Pullet length Tarsus length Corn C C C 117.1 179.8 60.3 73.0 47.9 63.1 C C M 112.3 179.1 60.4 74.1 49.1 63.7 C M M 117.3 183.9 64.2 78.4 49.0 64.4 M M M 125.2 194.8 64.3 78.7 49.4 64.9 Wheat C C C 114.1 178.3 60.6 74.4 46.1 60.2 C C M 114.5 178.3 60.9 73.3 48.6 64.8 C M M 114.9 181.0 62.2 75.4 49.0 65.1 M M M 118.2 185.6 62.2 76.0 48.8 65.5 Main effects Cereal Corn 118.0 184.4 62.3 76.1 48.9 64.0 Wheat 115.4 180.8 61.5 74.8 48.1 63.9 Feed form C C C 115.5b 179.0b 60.5 73.7b 47.0b 61.7b C C M 113.4b 178.8b 60.7 73.7b 48.9a 64.2ab C M M 116.1ab 182.5ab 63.2 76.9ab 49.0a 64.8ab M M M 121.7a 190.2a 62.2 77.4a 49.1a 65.2a SEM4 6.61 10.75 8.955 3.831 1.74 3.60 Probability Cereal 0.106 0.159 0.693 0.158 0.073 0.870 Feed form 0.003 0.007 0.653 0.004 0.014 0.020 Feed form*cereal 0.232 0.635 0.952 0.291 0.478 0.323 1C=crumble; M=mash. 2Measured from the tip of the beak to the end of the longest phalanx. 3Measured with a caliper above the spur. 4 SEM (9 replicates of 2 pullets each per treatment).