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595 Advances in Environmental Biology, 5(4): 595-604, 2011 ISSN 1995-0756 This is a refereed journal and all articles are professionally screened and reviewed ORIGINAL ARTICLE Evaluation of Growth Performance of Broiler Chicks Fed with Diet Containing Chickpea Seeds Supplemented with Exogenous Commercial Enzymes Mehran Torki Department of Animal Science, Agriculture Faculty, Razi University Imam Avenue, Kermanshah, Iran, Postal Code: 6715685418. Mehran Torki: Evaluation of Growth Performance of Broiler Chicks Fed with Diet Containing Chickpea Seeds Supplemented with Exogenous Commercial Enzymes ABSTRACT This experiment was carried out to determine the effects of diet inclusion of chickpea seeds, as an alternative protein ingredient, with or without two commercial enzyme products on performance of broiler chicks. A total of 396 day-old Cobb broiler chicks were randomly distributed in 36 floor pens of 11 birds each. Six replicates were allocated to one of the six iso-energetic and iso-nitrogenous experimental diets. Chickpea were included in corn-soybean based diets at 100 g/kg as a partial replacement of soybean meal. Experimental diets were: 1- a corn-soybean based diet; 2- a low phosphorous corn-soybean based diet supplemented by 0.2 g/kg Ronozyme® (phytase activity); 3- a corn-soybean based diet supplemented by 0.4 g/kg Hemicell® (βmannanase activity); 4- a corn-soybean-chickpea based diet; 5- a low phosphorous corn-soybean-chickpea based diet supplemented with Ronozyme; 6- a corn-soybean-chickpea based diet supplemented by Hemicell. Starter, grower and finisher diets were offered to birds from 0 to 21 d, 22 to 42 d and 43 to 49 d of age, respectively. All experimental data were subjected to the GLM procedure of SAS as a complete randomized design. All statements of significance are based on a probability of less than 0.05. The mean values were compared by Duncan's Multiple Range Test. Dietary β-mannanase supplementation increased BWG of chicks fed the chickpea-included diet, but the effect of β-mannanase on BWG was not statistically significant in broilers fed the soybean meal-based diets. In finishing period, β-mannanase addition to chickpea-included diets improved FCR. Performance of birds fed low-P diet supplemented by phytase compared to chicks fed the soybean- or chickpea-included control diets had no statistically significant difference. From the results of this study, it can be concluded that chickpea can be included in broiler diet up to 10% with no adverse effect on bird performance. In addition, supplementing chickpea-included diets with Hemicell had beneficial effects on broiler performance in terms of FCR. Key words: chickpea, phytase, β-mannanase, enzyme, broiler chicks. Introduction Chickpea (Cicer arietinum L.; Family Leguminosae) is an ancient crop that has been grown in India, tropical, sub-tropical and temperate regions such as the Middle East and parts of Africa for many years. Chickpea is the 2nd most important pulse crop in the world, accounting for 14% [26] of world pulse production [53]. Chickpea seed has 38-59% carbohydrate, 3% fiber, 4.8-5.5% oil, 3% ash, 0.2% calcium, and 0.3% phosphorus. Digestibility of its protein varies from 76-78% and its carbohydrate from 57-60% [33,54]. Raw whole seeds contain per 100 g: 357 calories, 4.5-15.69% moisture, 0.8-6.4 % fat, 2.1-11.7 g fiber, 2-4.8 g ash, 140-440 mg Ca, 190-382 mg P, 9 mg Corresponding Author Mehran Torki, Department of Animal Science, Agriculture Faculty, Razi University Imam Avenue, Kermanshah, Iran, Postal Code: 6715685418. E-mail: [email protected] Adv. Environ. Biol., 5(4): 595-604, 2011 Fe, 0-225 μg β-carotene equivalent, 0.21-1.1 mg thiamin, 0.12-0.33 mg riboflavin, and 1.3-2.9 mg niacin [32]. The limiting amino acid concentrations are 0.52 for methionine, 1.45 for lysine and cystine, 0.71 for threonine and 0.16 for tryptophan [56]. The amino acid composition of seeds with 19.5% protein, 5.5% oil is (per 16 g N): 7.2 g lysine, 1.4 g methionine, 8.8 g arginine, 4.0 g glycine, 2.3 g histidine, 4.4 g isoleucine, 7.6 g leucine, 6.6 g phenylalanine, 3.3 g tyrosine, 3.5 g threonine, 4.6 g valine, 4.1 g alanine, 11.7 g aspartic acid, 16.0 g glutamic acid, 0.0 g hydroxyproline, 4.3 g proline, and 5.2 g serine [32,56]. Peas contain between 220 and 270 g/Kg crude protein and have an amino acid profile which is relatively well balanced [13] although like many grain legumes, the protein is marginal in the sulphur amino acids [42]. Although the protein concentration in pea is lower than lupins, soybeans or oilseed meals, the metabolisable energy (ME) is generally higher [22]. Although most of chickpea is produced for human consumption, a stock-feed market would provide a secondary market for reject grain and for excess production which is likely to occur occasionally in rapidly expanding industry, while providing the livestock industries with alternative sources of protein and energy-rich foods. To date the poultry industry has been reluctant to use these products in broiler diets due to suspected presence of anti-nutritional factors and very limited information on possible inclusion levels of untreated grains. Like other legumes, chickpea seeds contain varieties of anti-nutritional factors such as protease and amylase inhibitors, lectins, polyphenols and oligosaccharides [59,15]. Increasing the proportion of chickpea seed in the broiler diet negatively influenced body weight gain, food intake and food efficiency (Farrel 1999; 55 Pisulewski, 2000). Tentative recommendations to be made to industry for inclusion rate of chickpeas 10 g/kg. Non-starch polysaccharides (NSP) are complex high molecular weight carbohydrates found in the structure of plant cell walls and reduce nutrient bioavailability in poultry production [3,20,4,,17]. The NSP include various fiber types such as lignin, βglucans, arabinoxylans (pentosans), uronic acid, galactose, and mannose in poultry feedstuffs [2]. βMannan and its derivatives are integral components of cell walls in all legumes [48]. Certain protein concentrates, especially palm kernel meal, copra meal, and guar meal, are among the feedstuffs rich in glucomannans and galactomannans [2,57,14]. Several studies have demonstrated the negative effects of dietary β-mannan found in palm kernel meal, copra meal, guar gum, and guar meal [29]. βMannan is a linear polysaccharide composed of repeating β-1-4 mannose and 1-6 galactose and 596 glucose units attached to β-mannan backbone. Phytic acid is an ester of cyclic alcohol inositol with 6 phosphates. Its salt, phytate, is the major forms P [47] and occur mainly in seeds as mixed salts of magnesium and calcium [51]. Phytin is a collective term for this mixed salt [40] and it constitutes up to 3% of many of the oilseeds and cereals used in animal feeds. The inability of poultry to utilize phytate P, due to lack of endogenous phytase, results in the addition of inorganic feed P to poultry diets in order to meet the P requirements of poultry. However, P is the third most expensive nutrient in poultry diets after energy and protein [7] and phytate P passed out in animal excreta leading to accumulation of P in soils and subsequently its entry into surface and ground waters and sparking off major environmental concerns [34] . The poor digestive utilization of phytic acid P by monogastric animals and its consequences on diet cost, environment and digestibility of minerals and proteins have led to extensive research efforts directed toward improving phytate digestion [25]. Enzyme supplementation of poultry rations has been widely reviewed and well documented to improve efficiency of converting feedstuffs into broiler tissue [3,28,12, Fuente et al., 1995; Bedford and Morgan, 1996;36]18, as well as growth performance and nutrient digestion in poultry [5,37,1 58]. Beneficial effects of exogenous enzymes in broiler diets containing oilseeds (Cowiesson et al., 1999) and lupins [30,39] have also been reported. Hemicell® is a fermentation product of Bacillus lentus. It contains high amounts of β-Mannanase that degrade β-mannan in feed. β-Mannanase has been shown to improve feed conversion of broilers [57]. Broilers fed low-energy diets supplemented with βMannanase performed slightly better than broilers fed high-energy diets without enzyme [38]. Phytase (myo-inositol-hexakisphosphohydrolase) degrades phytate to yield inositol monophosphate and orthophosphate via inositol penta to monophosphates as intermediary products [35]. It has been well documented that phytase improves P utilization in poultry [52,49,24, 31,54,10 41, 44]. Further studies have suggested that microbial phytase can have beneficial effects on amino acid digestibility in poultry [8,50,45,]. Although the benefits of the addition of exogenous enzymes to wheat, rye, barley, oilseeds and lupins have well documented, whether the benefits of phytase and β–mannanase will also be demonstrable in chickpea-based diets is not as evident. The present experiment was conducted in order to investigate the effects of addition of exogenous enzymes to chickpea included diets on performance, slaughtered traits, antibody response and litter pH and moisture of growing broiler chicks. Adv. Environ. Biol., 5(4): 595-604, 2011 Materials and methods All procedures used in this six-week experiment were approved by the Animal Ethics Committee of Razi University and complied with the “Guidelines for the Care and Use of Animals in Research". A total of 396 unsexed day-old Cobb broiler chicks were obtained from a local hatchery, weighed and randomly allocated to 36 pens. Eleven birds placed in each pen. Any birds showing signs of ill health, injury or being in poor condition were discarded. There were a total of six iso-energetic and isonitrogenous experimental diets, each replicated 6 times. Two commercial exogenous dietary enzyme preparations which were used in this study, Hemicell® and Ronozyme P 5000® (CT: Coated Thermotolerant), have β-mannanase and phytase activity, respectively. The Hemicell was provided by Chem Gen Co., Ltd., and the activity of β-mannanase was 165 × 106 U/kg (inclusion dose: 400 g/t feed). Ronozyme P 5000 was provided by DSM Nutritional Products Ltd, Switzerland (inclusion dose: 150 g/t feed). Chickpea was included in corn-soybean based diets at 100 g/kg as a partial replacement of soybean meal. 597 Experimental diets were: 1- a corn-soybean based diet; 2- a low phosphorous corn-soybean based diet supplemented with 0.2 g/kg Ronozyme® (phytase activity); 3- a corn-soybean based diet supplemented by 0.4 g/kg Hemicell® (β-mannanase activity); 4, a corn-soybean-chickpea based diet; 5- a low phosphorous corn-soybean-chickpea based diet supplemented with Ronozyme; 6- a corn-soybeanchickpea based diet supplemented by Hemicell. The levels of dietary Ca and available P in control and low P groups were 0.89, 0.40 and 0.89, 0.24 in starter, 0.82, 0.32 and 0.82, 0.19 in grower, and 0.74, 0.28 and 0.74, 0.17 in finisher diets, respectively. Starting (ME=2850 Kcal/kg and 20.48 % CP), growing (ME=2900 Kcal/kg and 18.13 % CP), and finishing (ME=2950 Kcal/kg and 16.59 % CP) diets were fed from 0-21, 22-42, and 43-49 days of age, respectively. Tables 1-3 show the experimental diets. Water and feed was available ad libitum throughout the study. Body weights and feed intake were monitored by the end of each rearing periods (d 1, 21, 42, and 49). Table 1: Composition of experimental diets for starting period (0-21 days) Corn-soybean meal-based diets Corn-soybean meal-chickpea-based diets -------------------------------------------------------------------------------------------------------------------------Ingredients No enzyme Phytase β-mannanase No enzyme Phytase β-mannanase -------------------------------------------------------------------------------------------------------------------------------------------------------------------------g / 100 g diet Corn 59.96 59.84 59.97 55.59 55.47 55.59 Chickpea 10 10 10 Soybean meal 33.93 33.7 33.95 29.86 29.63 29.88 Wheat bran 1.71 2.4 1.64 0.07 0.75 Sunflower oil 0.55 0.55 0.55 0.55 0.55 0.55 DCP1 1.37 0.51 1.37 1.47 0.6 1.47 Oyster shell 1.25 1.75 1.25 1.2 1.7 1.2 Salt 0.5 0.5 0.5 0.5 0.5 0.5 Mineral premix2 0.25 0.25 0.25 0.25 0.25 0.25 Vitamin premix3 0.25 0.25 0.25 0.25 0.25 0.25 Lysine-Hcl 0.1 0.1 0.1 0.12 0.13 0.12 DL-Methionine 0.13 0.13 0.13 0.14 0.14 0.14 Phytase4 0.02 0.02 0.04 0.04 β-Mannanase5 Calculated analysis ME (Kcal/kg) 2850 2850 2850 2850 2850 2850 Crude protein % 20.48 20.48 20.48 20.48 20.48 20.48 Ether extract % 3.1 3.11 3.1 2.98 2.99 2.98 Crude fiber 3.88 3.94 3.88 3.87 3.93 3.86 Calcium 0.89 0.89 0.89 0.89 0.89 0.89 Available phosphorous 0.4 0.24 0.4 0.4 0.24 0.4 Lysine 1.15 1.15 1.15 1.21 1.21 1.21 Methionine 0.44 0.44 0.44 0.44 0.44 0.44 Methione + Cystein 0.78 0.78 0.78 0.77 0.77 0.77 1 DCP: Dicalcium phosphate; 2 The mineral premix supplied per kilogram of diet: zinc, 120 mg; manganese, 120 mg; iron, 80 mg; copper, 10 mg; iodine, 2.5 mg; cobalt, 1 mg; 3The vitamin premix supplied per kilogram of diet: vitamin A, 12,500 IU; vitamin D3, 4,000 IU; vitamin E, 66 IU; vitamin B12, 39.6 µg; riboflavin, 13.2 mg; niacin, 165 mg; D-pantothenic acid, 44 mg; menadione, 0.4 mg; folic acid,3.3 mg; pyridoxine, 13.75 mg; thiamin, 6.66 mg; D-biotin, 0.28 mg, 4Ronozyme, 5Hemicell Adv. Environ. Biol., 5(4): 595-604, 2011 598 Table 2: Composition of experimental diets for growing period (21-42 days). Corn-soybean meal-based diets Corn-soybean meal-chickpea-based diets -------------------------------------------------------------------------------------------------------------------------Ingredients No enzyme Phytase β-mannanase No enzyme Phytase β-mannanase -------------------------------------------------------------------------------------------------------------------------------------------------------------------------g / 100 g diet Corn 67.6 67.5 67.61 63.23 63.13 63.24 Chickpea 10 10 10 Soybean meal 27 26.83 27.03 22.93 22.75 22.95 Wheat bran 1.89 2.42 1.81 0.24 0.78 0.17 DCP1 0.99 0.31 0.99 1.09 0.4 1.09 Oyster shell 1.32 1.72 1.32 1.28 1.67 1.28 Salt 0.5 0.5 0.5 0.5 0.5 0.5 Mineral premix2 0.25 0.25 0.25 0.25 0.25 0.25 Vitamin premix3 0.25 0.25 0.25 0.25 0.25 0.25 Lysine-Hcl 0.15 0.15 0.15 0.17 0.17 0.17 DL-Methionine 0.05 0.05 0.05 0.07 0.07 0.07 Phytase4 0.02 0.02 0.04 0.04 β-Mannanase5 Calculated analysis ME (Kcal/kg) 2900 2900 2900 2900 2900 2900 Crude Protein % 18.12 18.12 18.12 18.12 18.12 18.12 Ether extract % 2.84 2.85 2.84 2.72 2.73 2.72 Crude fiber 3.58 3.63 3.58 3.57 3.62 3.57 Calcium 0.81 0.81 0.81 0.81 0.81 0.81 Available phosphorous 0.32 0.19 0.32 0.32 0.19 0.32 Lysine 1.02 1.02 1.02 1.08 1.08 1.08 Methionine 0.34 0.34 0.34 0.34 0.34 0.34 Methione + Cystein 0.65 0.65 0.65 0.64 0.64 0.64 1 DCP: Dicalcium phosphate; 2The mineral premix supplied per kilogram of diet: zinc, 120 mg; manganese, 120 mg; iron, 80 mg; copper, 10 mg; iodine, 2.5 mg; cobalt, 1 mg; 3The vitamin premix supplied per kilogram of diet: vitamin A, 12,500 IU; vitamin D3, 4,000 IU; vitamin E, 66 IU; vitamin B12, 39.6 µg; riboflavin, 13.2 mg; niacin, 165 mg; D-pantothenic acid, 44 mg; menadione, 0.4 mg; folic acid,3.3 mg; pyridoxine, 13.75 mg; thiamin, 6.66 mg; D-biotin, 0.28 mg, 4Ronozyme, 5Hemicell Table 3: Composition of experimental diets for finishing period (42-49 days). Corn-soybean meal-based diets Corn-soybean meal-chickpea-based diets -------------------------------------------------------------------------------------------------------------------------Ingredients No enzyme Phytase β-mannanase No enzyme Phytase β-mannanase Corn 71.91 71.83 71.92 67.54 67.46 67.55 Chickpea 10 10 10 Soybean meal 22.91 22.76 22.93 18.85 18.7 18.87 Wheat bran 2.04 2.5 1.96 0.39 0.85 0.31 DCP1 0.81 0.22 0.81 0.91 0.31 0.91 Oyster shell 1.25 1.59 1.25 1.2 1.55 1.2 Salt 0.5 0.5 0.5 0.5 0.5 0.5 Mineral premix2 0.25 0.25 0.25 0.25 0.25 0.25 Vitamin premix3 0.25 0.25 0.25 0.25 0.25 0.25 Lysine-Hcl 0.06 0.06 0.06 0.08 0.08 0.08 DL-Methionine 0.02 0.02 0.02 0.03 0.03 0.03 Phytase4 0.02 0.02 ?-Mannanase5 0.04 0.04 Calculated analysis ME (Kcal/kg) 2950 2950 2950 2950 2950 2950 Crude protein % 16.59 16.59 16.59 16.59 16.59 16.59 Ether extract % 2.98 2.99 2.97 2.86 2.87 2.86 Crude fiber 3.41 3.45 3.4 3.4 3.43 3.39 Calcium 0.74 0.74 0.74 0.74 0.74 0.74 Available phosphorous 0.28 0.28 0.28 0.28 0.28 0.28 Lysine 0.86 0.86 0.86 0.91 0.91 0.91 Methionine 0.29 0.29 0.29 0.29 0.29 0.29 Methione + Cystein 0.58 0.58 0.58 0.57 0.57 0.57 1 DCP: Dicalcium phosphate; 2The mineral premix supplied per kilogram of diet: zinc, 120 mg; manganese, 120 mg; iron, 80 mg; copper, 10 mg; iodine, 2.5 mg; cobalt, 1 mg; 3The vitamin premix supplied per kilogram of diet: vitamin A, 12,500 IU; vitamin D3, 4,000 IU; vitamin E, 66 IU; vitamin B12, 39.6 µg; riboflavin, 13.2 mg; niacin, 165 mg; D-pantothenic acid, 44 mg; menadione, 0.4 mg; folic acid,3.3 mg; pyridoxine, 13.75 mg; thiamin, 6.66 mg; D-biotin, 0.28 mg, 4Ronozyme, 5Hemicell Carcass measurements were taken on one randomly selected bird per each pen on day 49 of age. Feed was withdrawn 3 hours prior to slaughter. Birds were manually hung on shackles, stunned with an electric knife, bled for 3 min after the jugular vein was severed. Hocks and heads were removed manually. Viscera and abdominal fat were then removed manually. Carcass, various viscera parts and abdominal fat weights were recorded. Breast, wings, thighs and drumsticks were removed from the Adv. Environ. Biol., 5(4): 595-604, 2011 carcass, weighed and recorded. Weights for each variable measured, were expressed as relative to live BW at processing. On d 35, primary antibody response to Newcastle disease virus was measured in one randomly selected bird from each pen. Birds were immunized on day 15. At d 35, birds were bled via the wing vein to collect serum used or evaluating primary antibody response. To measure litter pH, a 200-g litter sample was collected from various places of each pen when the birds reached 47 d of age. The pH-meter was at first calibrated by buffer=10 solution then pH determined by stirring 50 g of litter into 500 ml of deionizeddistilled H2O for 5 min and the H+ concentration of the resulting solution measured. In addition, the pH meter was calibrated by buffer=10 solution after each 16 testing. Litter moisture was determined gravimetrically by drying the samples at 70oC until the weight of samples reached to a constant level. All experimental data were subjected to the GLM procedure of SAS as a complete randomized design. All statements of significance are based on a probability of less than 0.05. The mean values were compared by Duncan's Multiple Range Test. Results and discussion Body weight (BW) means and standard deviations of chicks fed on experimental diets on days 21, 42 and 49 are shown in table 4. There was no statistically significant difference between dietary groups in BW on day 21 of age. Chicks BW on days 42 and 49 of age was affected by dietary treatment. On day 42, chicks fed on phytase-supplemented low P soybean-based diets had significantly higher BW than those fed on phytase-supplemented low P chickpea-included diets (P < 0.05). There was no statistically significant difference in chick BW between other dietary groups (P > 0.05). Chicks fed β-mannanase supplemented chickpea included diets did have the highest BW on day 49 of age that was significantly higher than BW of birds fed chickpeaincluded diets (P < 0.05). No significant difference was seen in BW between birds fed corn-soybean meal-based diets (P > 0.05) Body weight gain (BWG), feed intake (FI) and feed conversion ratio (FCR) are provided in table 5. Feeding low P, phytase supplemented diet decreased chicks BWG. BWG of chicks fed the low P, phytase supplemented chickpea-included diet compared to birds fed the low P, phytase supplemented soybeanbased diet was lower on average of 21-42 days of age (P<0.05). The highest and lowest BWG during days 42-49 of age was seen in birds fed the βmannanase supplemented chickpea-included and low P phytase-supplemented soybean meal diets, respectively (P<0.05). Dietary treatment did have 599 significant effect on FI in growing period. Birds fed the chickpea-included diets with no supplementary enzyme fed less than birds fed the soybean mealbased diets supplemented by β-mannanase (P<0.05). Enzyme addition increased FI of chicks fed on chickpea-included diets. Dietary treatment significantly affected FCR during finishing period (P<0.05), but there was no significant difference in FCR between dietary groups during starting and growing periods (P>0.05). In finishing period, βmannanase addition to chickpea-included diets improved FCR which was better than FCR in low P soybean-based diets (P<0.05). Performance of birds fed the low P, phytase supplemented diet had no statistically significant difference compared to chicks fed the soybean- or chickpea-included control diets. In other studies increasing the proportion of chickpea seed in the broiler diets negatively influenced body weight gain, food intake and food efficiency (Farrel 1999; 55]. McNaughten et al. (1998) and Odetallah et al. (2002), which showed that the addition of β-mannanase to corn-soybean diets improved performance of birds fed soybeanbased diets. Viveros et al. [55] who formulated diets to contain 75 and 150 g/kg of raw and autoclaved Desi chickpea seeds, weight gain and FI of the chicks given Desi chickpea diets was significantly reduced compared to those fed the control diet. Increasing the proportion of seed in the diet negatively influenced BWG, FI and FCR. Moreover, a significant increment in the relative weights of liver and pancreas and in the relative lengths of duodenum, ileum and ceaca was observed when the concentration of chickpea seeds in the diets was increased. Feeding autoclaved seeds significantly increased the BWG and the FI. However, FCR was not modified by the autoclaving. Relative weights of gizzard and liver and relative lengths of ileum were decreased significantly by the inclusion of autoclaved desi chickpea in the diet. These differences might be due to the inclusion level or chickpea varieties used. Christodoulou et al. [19] reported that the replacement of soybean meal with extruded chickpeas in broiler turkey diets at inclusion levels (400, 600 and 800 kg/t of diet) decreased body weight and increased feed conversion ratio compared to the control. Brenes et al [11] who evaluated the inclusion of different concentrations (0, 100, 200 and 300 g kg-1) of raw and extruded chickpeas on performance of growing broiler chickens reported that Increasing chickpea content in the diet did not affect weight gain, feed consumption and feed to gain ratio. Carcass parameters are demonstrated in tables 6 and 7. Dietary treatment had significant effect on relative weights of heart and duodenum to live body weight. In other reports, the relative weights of the pancreas, liver and gizzard and the relative lengths of duodenum, jejunum, ileum and caeca were increased Adv. Environ. Biol., 5(4): 595-604, 2011 and amylase and trypsin activities and AMEn were reduced (Farrel 1999; 55]. Relative pancreas and liver weights, and relative lengths of duodenum, jejunum and ceca were significantly increased in response to increasing chickpea concentration in the diet [11]. Christodoulou et al. [19] showed that carcass yield traits of broiler turkeys were not affected by feeding diets with increasing levels (400, 600 and 800 kg/t of diet) of extruded chickpeas. 600 Antibody response to Newcastle disease virus did not significantly affected by dietary treatment (table 8). It can be concluded that partial replacement of soybean meal by chickpea in broiler diets would have no detrimental effects on BWG, FI and FCR. Supplementing chickpea-included diets with Hemicell had beneficial effects on broiler performance in terms of FCR. Table 4: Body weight (g /bird) of broiler chicks fed corn-soybean meal or corn-soybean meal-chickpea-based diets with or without enzyme supplementation Experimental diets Enzyme Body weight (g/chick) -------------------------------------------------------------------------------------------------------21 42 49 Soybean No enzyme 640.93 ± 16.02 1872.77 ± 70.14ab 2283.08 ± 102.42ab Soybean-low P Phytase 626.93 ± 24.74 1910.71 ± 72.11a 2257.03 ± 123.12ab Soybean ?-Mannanase 617.78 ± 37.38 1815.99 ± 141.19ab 2243.51 ± 96.41ab Chickpea No enzyme 632.73 ± 36.34 1845.01 ± 83.41ab 2228.23 ± 155.97b Chickpea-low P Phytase 626.52 ± 15.49 1795.60 ± 117.56b 2238.23 ± 111.62b Chickpea β-Mannanase 638.86 ± 15.32 1909.42 ± 42.51ab 2378.83 ± 136.46a Pooled SEM 4.24 16.21 20.81 Means ± standard deviation, a-b Means within a column (within main effects) with no common superscript differ significantly (P <0.05). Table 5: Growth rate (gain/bird/d), feed intake (feed/bird/d) and feed conversion ratio (FCR) of broiler chicks fed corn-soybean meal or corn-soybean meal-chickpea-based diets with or without exogenous enzyme supplementation. Diet Enzyme Body weight gain (g/chick/day) --------------------------------------------------------------------------------------------------------------------0-21 21-42 42-49 0-49 Soybean No enzyme 28.38 ± 0.76 58.09 ± 3.44ab 64.7 ± 11.23ab 46.37 ± 1.45ab Soybean-low P Phytase 27.69 ± 1.18 61.07 ± 2.96a 51.74 ± 4.80b 45.32 ± 1.77ab Soybean ?-Mannanase 27.28 ± 1.79 56.73 ± 6.06ab 56.29 ± 5.83ab 44.99 ± 1.77ab Chickpea No enzyme 27.99 ± 1.73 57.88 ± 3.56ab 57.88± 11.89ab 44.88 ± 2.46b Chickpea-low P Phytase 27.69 ± 0.74 55.20 ± 5.34b 62.91 ±14.39ab 44.58 ± 1.38b Chickpea β-Mannanase 28.28 ± 0.73 60.47 ± 1.87ab 67.03 ± 10.41a 47.44 ± 2.27a Pooled SEM 0.202 0.723 1.831 0.337 Diet Enzyme Feed intake (g/chick/day) ----------------------------------------------------------------------------------------------------------------0-21 21-42 42-49 0-49 Soybean No enzyme 47.88 ± 3.55 114.27± 3.29ab 179.67± 13.62 95.37 ± 3.14ab Soybean-low P Phytase 49.43 ± 1.51 118.20± 4.51ab 180.74 ± 8.05 96.72 ± 1.84ab Soybean ?-Mannanase 49.84 ± 1.12 120.31± 5.63a 179.46 ± 9.11 97.94 ± 3.45a Chickpea No enzyme 47.68 ± 1.01 111.59 ± 4.11b 179.11 ± 8.29 92.91 ± 4.85b Chickpea-low P Phytase 47.98 ± 1.63 114.65± 2.19ab 181.54 ± 9.40 95.07 ± 1.07ab Chickpea β-Mannanase 48.78 ± 0.43 116.29± 4.09ab 181.74 ± 8.53 95.88 ± 2.02ab Pooled SEM 0.310 1.022 1.511 0.531 Diet Enzyme Feed conversion ratio (g: g) ------------------------------------------------------------------------------------------------------------------0-21 21-42 42-49 0-49 Soybean No enzyme 1.69 ± 0.16 1.97 ± 0.09 2.85 ± 0.49ab 2.06 ± 0.11ab Soybean-low P Phytase 1.79 ± 0.79 1.94 ± 0.11 3.51 ± 0.25a 2.14 ± 0.08ab Soybean ?-Mannanase 1.83 ± 0.09 2.14 ± 0.27 3.20 ± 0.17ab 2.18 ± 0.21a Chickpea No enzyme 1.71 ± 0.12 1.94 ± 0.27 3.19 ± 0.58ab 2.08 ± 0.15ab Chickpea-low P Phytase 1.77 ± 0.07 2.09 ± 0.18 3.01 ± 0.64ab 2.13 ± 0.06ab Chickpea β-Mannanase 1.73 ± 0.05 1.93 ± 0.11 2.76 ± 0.41b 2.03 ± 0.12b Pooled SEM 0.018 0.032 0.082 0.018 Means ± standard deviation, a-b Means within a column (within main effects) with no common superscript differ significantly (P <0.05). Table 6: Carcass yield parameters of broiler chicks fed diets based on corn-soybean meal or corn-soybean meal-chickpea with or without exogenous enzymes supplementation. Diet Enzyme % of live body weight ----------------------------------------------------------------------------------------------------Viscera Skin & Feather Abdominal fat Soybean No enzyme 5.86 ± 3.59 14.02 ± 0.81 2.11 ± 0.65 Soybean-low P Phytase 6.11 ± 1.55 13.79 ± 0.69 2.04 ± 0.75 Soybean β-Mannanase 6.18 ± 1.51 13.49 ± 1.32 2.09 ± 0.76 Chickpea No enzyme 6.07 ± 2.51 13.5 ± 1.19 2.10 ± 0.40 Chickpea-low P Phytase 5.96 ± 4.60 13.57 ± 1.48 1.94 ± 0.34 Chickpea β-Mannanase 6.06 ± 0.74 13.53 ± 1.11 2.03 ± 0.56 Adv. Environ. Biol., 5(4): 595-604, 2011 Table 6: Continue. Pooled SEM Diet 601 0.536 0.178 0.111 % of live body weight ------------------------------------------------------------------------------------------------------Thigh Drumstick Heart Soybean No enzyme 8.36 ± 1.00 9.22 ± 0.78 0.51 ± 0.05ab Soybean-low P Phytase 8.75 ± 0.63 9.46 ± 0.82 0.58 ± 0.09ab Soybean ?-Mannanase 8.53 ± 1.37 8.82 ± 0.75 0.49 ± 0.04ab Chickpea No enzyme 8.46 ± 0.74 9.71 ± 0.41 0.59 ± 0.06a Chickpea-low P Phytase 8.85 ± 0.94 9.29 ± 0.66 0.59 ± 0.11a Chickpea β-Mannanase 8.51 ± 6.62 9.27 ± 0.93 0.48 ± 0.11b Pooled SEM 0.146 0.123 0.014 Means ± standard deviation, a-b Means within a column (within main effects) with no common superscript differ significantly (P <0.05). Enzyme Table 7: Carcass yield parameters of broiler chicks fed corn-soybean meal or corn-soybean meal-chickpea-based diets with or without exogenous enzymes supplementation Diet Enzyme % of live body weight ----------------------------------------------------------------------------------------------------Liver Gizzard Pancreas Soybean No enzyme 2.64 ± 0.435 2.59 ± 0.61 0.20 ± 0.02 Soybean-low P Phytase 2.31 ± 0.38 2.65 ± 0.53 0.20 ± 0.06 Soybean β-Mannanase 2.52 ± 0.49 2.40 ± 0.34 0.21 ± 0.69 Chickpea No enzyme 2.23 ± 0.46 2.57 ± 0.12 0.21 ± 0.22 Chickpea-low P Phytase 2.40 ± 0.32 2.60 ± 0.758 0.24 ± 0.03 Chickpea β-Mannanase 2.31 ± 0.19 2.58 ± 0.26 0.22 ± 0.07 Pooled SEM 0.064 0.076 0.008 Diet Enzyme % of live body weight ------------------------------------------------------------------------------------------------------Deodenum Jeojenum Ileum Cecae Soybean No enzyme 0.71 ± 0.02ab 1.91 ± 0.59 1.67 ± 0.43 0.64 ± 0.23 Soybean-low P Phytase 0.68 ± 0.12ab 1.57 ± 0.42 1.58 ± 0.575 0.63 ± 0.19 Soybean β-Mannanase 0.64 ± 0.10a 1.90 ± 0.46 1.48 ± 0.33 0.55 ± 0.06 Chickpea No enzyme 0.63 ± 0.08b 1.95 ± 0.52 1.68 ± 0.48 0.50 ± 0.05 Chickpea-low P Phytase 0.69 ± 0.11ab 1.61 ± 0.59 1.47 ± 0.63 0.58 ± 0.13 Chickpea β-Mannanase 0.67 ± 0.13ab 1.77 ± 0.47 1.70 ± 0.52 0.62 ± 0.28 Pooled SEM 0.016 0.083 0.081 0.029 Means ± standard deviation, a-b Means within a column (within main effects) with no common superscript differ significantly (P <0.05). Table 8: Humoral immune response against Newcastle disease virus of broiler chicks fed corn-soybean meal or corn-soybean meal-chickpea-based diets with or without exogenous enzymes supplementation Diet Enzyme Antibody titers (log 2) ----------------------------------------------------------------------------------------------------Day 15 of age (day of immunization) Day 35 of age (primary Ab response) Soybean No enzyme 2.75 ± 1.29 8.00 ± 1.16 Soybean-low P Phytase 2.60 ± 1.52 7.33 ± 1.16 Soybean β-Mannanase 2.67 ± 0.58 7.20 ± 1.30 Chickpea No enzyme 2.50 ± 0.58 5.80 ± 2.95 Chickpea-low P Phytase 3.00 ± 0.01 6.80 ± 0.45 Chickpea β-Mannanase 2.20 ± 0.90 6.50 ± 1.52 MSE 0.176 0.312 Means ± standard deviation, References 1. 2. 3. Almirall, M., M. Francesch, A.M. PerezVendrell, J. Brufau and E. Esteve-Garcia, 1995. 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