Laying Performance and Egg Quality of Japanese Quails Fed Diets Containing Castor Meal and Enzyme Complex

The objective of this study was to evaluate the effect of diets formulated with corn plus soybean meal (CSM) or with 21% autoclaved castor meal (ACM), with the addition of two enzyme complexes (EC1 and EC2), on the performance and egg quality of laying quail. Two hundred and sixteen quails were selected by egg production and weight uniformity and distributed in a completely randomized design with six treatments (in a 2 × 3 cross-factorial arrangement) and six replicates with six birds each. Three CSM and three ACM diets supplemented withEC1, EC2, or unsupplemented were established. The trial lasted three cycles of 21 days. Feed intake, laying rate, egg mass, feed conversion per egg mass, and feed conversion per dozen eggs did not differ. The use of ACM diet reduced egg (EW), yolk (YW), and eggshell (SW) weights, egg specific gravity and increased yolk color. However, EW and YW were similar among quails fed diets containing CSM and ACM when supplemented with CE1.The use of enzyme complex containing xylanase, β-glucanase, and phytase is recommended when 21% autoclaved castor mealis included in the diet of laying quails.

formulated with corn plus soybean meal (CSM) or with 21% autoclaved castor meal (ACM), with the addition of two enzyme complexes (EC1 and EC2), on the performance and egg quality of laying quail.Two hundred and sixteen quails were selected by egg production and weight uniformity and distributed in a completely randomized design with six treatments (in a 2 × 3 cross-factorial arrangement) and six replicates with six birds each.Three CSM and three ACM diets supplemented withEC1, EC2, or unsupplemented were established.The trial lasted three cycles of 21 days.Feed intake, laying rate, egg mass, feed conversion per egg mass, and feed conversion per dozen eggs did not differ.The use of ACM diet reduced egg (EW), yolk (YW), and eggshell (SW) weights, egg specific gravity and increased yolk color.However, EW and YW were similar among quails fed diets containing CSM and ACM when supplemented with CE1.The use of enzyme complex containing xylanase, β-glucanase, and phytase is recommended when 21% autoclaved castor mealis included in the diet of laying quails.

IntRoduCtIon
With the development of the poultry industry, major advances have been achieved, providing alternatives to make feed compounding more efficient and economical.Enzymes have an effect on diet cost reduction and contribute to reducing antinutritional effects of ingredients, improving the diet digestibility and poultry performance (Olukosi et al., 2007), especially when alternative feedstuffs, such as castor (Ricinus communis L.), are used in the feed.
Castor (Ricinus communis L.) is one of the most traditional crops grown in the Brazilian northeast semi-arid region.After being pressed, castor seeds generate the castor cake, a high-protein and high-fiber solid extract residue.Its high protein content (30.9%, according to Matos Júnior et al. (2011) renders it an attractive alternative for animal feed, but the presence of toxic and allergenic compounds, such as ricin and2S albumins, have made its use impractical on a large commercial scale (Severino et al., 2012).However, according to Anandan et al. (2005), the heat and pressing treatment of the meal associated with chemical agents such as CaO or Ca(OH) 2 may inactivate ricin and the allergen compound CB-1A, identified by Youle & Huang (1978) as 2S albumins, in beans and castor seeds.
Defatted castor seed meal has a high concentration of insoluble fiber, which can reduce the nutrient digestibility as well as the performance of poultry (Santos et al., 2015).Based on Brazilian literature castor cake, expected average composition is 60% total carbohydrates, 10% non-fibrous carbohydrates, 50% neutral detergent fiber, 40% acid Laying Performance and Egg Quality of Japanese Quails Fed Diets Containing Castor Meal and Enzyme Complex detergent fiber, and 25% lignin.As stated by Costa et al. (2004), the insoluble fiber percentage in the castor cake and defatted castor meal can vary between 43 and 51%, given the processing variability and different castor varieties.The insoluble-fiber fraction has low digestibility in poultry, increases endogenous nutrient loss, and simultaneously reduces the presence of potentially metabolizable nutrients in the diet, thereby preventing the action of enzymes on the digest and reducing the energy concentration of the feed.The effects of fiber are associated with viscosity, which can increase as a result of the non-starch polysaccharide content, especially its β-glucan and arabinoxylan fractions (Tavernari et al., 2008).Increases in viscosity may reduce nutrient digestion in the small intestine (Jaroni et al., 1999).According to Chotinsky et al. (2015), the exact effects of viscosity have not been established, but possible mechanisms include reduced rates of diffusion of endogenous enzymes and nutritional substrates.
However, the use of exogenous enzymes can reduce the viscosity of the digesta (Bharathidhasan et al., 2009) as well as improve the nutrient utilization (Jaroni et al., 1999).
This study was conducted to evaluate the effect of diets formulated with corn plus soybean meal or with 21% autoclaved castor meal, plus the addition of two enzyme complexes, on performance and egg quality of laying quails.

MAteRIAl And MethodS
The experiment was conducted in Recife, Brazil 8°03'28" S latitude and 34°52'58" W longitude, at 9m (asl).Procedures involving animals were performed according to the Institutional Committee on Animal Use (license no.043/2012).Two hundred and sixteen quails of 39 days of age were housed in 36 galvanized-wire cages (33 × 25 × 20cm) in a room with temperature control set at 22°C.After reaching peak egg production, the selected quails were distributed in a completely randomized design with six treatments, six replicates, and six birds per experimental unit.The selection and distribution criteria were uniformity in body weight and production.At the beginning of the experiment, quails had an average weight of 160.1g and egg laying rate above 90%.The trial period lasted 63 days (21-day cycles) and quails received 17 h of light per day.Temperature was monitored once a day, using maximum and minimum thermometers.The quails were fed a mash diet twice daily, and water was provided ad libitum.
Castor meal was provided by the Biodiesel Unit located in Pesqueira-PE, Brazil.The castor meal was homogenized with 6% CaO; subsequently, water was added at a 1:1 ratio by weight with homogenization until forming uniform wet mixture.This mixture was placed in vats (to a height of 25cm) that were placed on the autoclave.The mixture was autoclaved at a pressure of 1.23kgf cm −2 , at 104 °C, for 90 min.Each of the vats placed in the autoclave had its upper part open and unobstructed, allowing the exit of steam.After processing, the autoclaved castor meal was sundried for later storage and milling.The procedures were adapted from recommendations of Anandan et al. (2005) and Cobianchi et al. (2012).
Because both EC are active in different substrates, given their different compositions, we evaluated their effects on the measured parameters so as not to individualize the isolated effect of the present enzymes.In the formulation of the diets (Table 2), we adopted the recommendations of nutritional requirements described by Silva & Costa (2009), while the ingredient composition was based on Rostagno et al. (2011).The level of autoclaved castor meal was established in an experiment performed previously in which we identified that laying quail performance declined if levels equal to or greater than 21% were used.
Diets were formulated using a single nutritional matrix, regardless of EC addition.This formulation procedure is named "over the top" (Scheideler et al., 2005), and we used it in this experiment to determine whether EC has the ability to repair possible negative effects from the use of ACM.

Laying Performance and Egg Quality of Japanese Quails Fed Diets Containing Castor Meal and Enzyme Complex
For performance evaluation, eggs were counted and weighed daily and feed leftovers and average egg weight were quantified weekly to determine egg production per day (laying rate, %), feed intake (FI, g day −1 ), egg weight (EW, g), egg mass (EM, g day −1 ), feed conversion per egg mass (FCEM, g of egg g of diet −1 ), and feed conversion per dozen eggs (FCDZ, g of diet 12 eggs −1 ).
Eggs were weighed daily and a representative sample of average weight of eggs, per experimental unit, was obtained from the last three days of each cycle to determine EW.After 63 days, mean values from 18 eggs per experimental unit were generated (2 eggs day −1 , 3 days cycle −1 , three cycles).Specific gravity was measured by immersing the egg in saline solutions; for this step, ten solutions were prepared in labeled cans with densities ranging from 1.050 to 1.100g cm −3 , with a gradual increase of 0.005g cm −3 , using distilled water at an average temperature of 22°C and salt (NaCl).
Haugh unit was established by the following procedure: after identification for EW determination, eggs were broken individually and contents were placed on a flat glass plate surface without inclination (on a stand with adjustable legs for leveling)to determine albumen height, which was measured to the nearest tenth of a millimeter using a digital caliper.Egg weight and albumen height (h, mm) values were used in the formula HU=100 log (h + 7.57 − 1.7 EW 0.37 ), according to Zita et al. (2013).To determine the percentages of shell and yolk, yolks were separated and weighed manually and shells were subsequently dried in a forced-air oven at 105°C for 24 h and weighed.Albumen weight was obtained as the difference between EW with SW and YW.
Shell thickness was assessed including membranes, using the same eggs broken to determine albumen quality.Shells were washed and then allowed to dry overnight at room temperature.After drying, eggshell thickness measurements were carried out on two different sites in the central cross-sectional eggshell area using a digital caliper.For colorimetry, two evaluators with normal vision used the DSM/Roche ® color fan (yolk fan) in which YC was compared with the color scale (ranging between one and fifteen) of the fan.
The main effect of factors and their interaction were assessed by analysis of variance.Means were compared by applying two Dunnett's test (at 5% probability) using as a reference either CSM diet (Dunnett's test 1) or ACM diet (Dunnett's test 2).Dunnett's test 1 allows for a comparison of all other treatments with CSM, while Dunnett's test 2 allows for a comparison of ACMEC1 and ACMEC2 with ACM.Statistical analyses were performed using SAS (Statistical Analysis System, version 9.4).Silva JCNS, Rabello CBV, Santos JS

ReSultS
During the trial period, the average recorded temperature was 27.4 ºC, with a minimum of 24.8 ºC and a maximum of 30.0 ºC.Average relative humidity was 68.3%.
The results are shows in Table 3 and 4. Most parameters showed no significant difference (p>0.05), and the following fit into this condition: FI, laying rate, EM, FCEM, FCDZ, ST, HU, AW, and percentages of yolk, albumen, and shell.
Significant effects were observed (p<0.05) for EW, YW, SG, YC, and SW, and these effects resulted from the use of diets with autoclaved castor meal, compared with the CSM diet, by Dunnett's test.Specific gravity was greater in eggs produced by quails receiving CSM diets as compared with eggs from quails fed ACM diets, conversely to YC results.
Shell weight differed significantly (P=0.0015) as affected by diet formulation.Because CaO was used in the castor meal processing, the amount of calcium added via limestone in ACM diets was reduced, and a reduction of 7% was observed in the SW of quail fed ACM diets.
Although it is considered a fibrous food, 21% autoclaved castor meal did not influence the evaluated parameters.A steady FI is the main parameter measured in animal trials to confirm adequate processing of castor cake and meal (Bueno et al., 2014).The mean performance values are comparable to those reported by Barreto et al. (2007) for the initial laying phase; however, they differ from those reported by Hemid et al. (2010), who evaluated the production of quails at the same age.
The parameters AFI, EM, FCEM, and FCDZ corroborate the data reported by Araujo et al. (2008), who did not find significant effects (p>0.05) on the evaluated variables using wheat meal and an enzyme complex in the diet of laying hens.However, the use of enzymatic complex in the study of Araujo et al. (2008) increased EW, while in the present study a slight decrease was detected in EW in comparing the 21% autoclaved castor meal diet and ACM plus EC2 in relation to CSM diet.This might have been because the diets with 21% autoclaved castor meal have a high crude fiber content (80.350 g kg −1 as fed) in comparison with CSM diet (30.060 g kg −1 as fed).According to Costa et al. (2004), the insoluble-fiber fraction has low digestibility in poultry, increases endogenous nutrient loss, and simultaneously reduces the presence of potentially metabolizable nutrients in the diet.The fibrous fraction (non-starch polysaccharide) may have promoted the increased viscosity in the digestive tract, which, according to Jaroni et al. (1999), may have reduced the digestibility of the nutrients.
Yolk, albumen, and shell percentages were close to those cited by Tolik et al. (2014), and the average ST (0.132mm) was 15% lower than that reported by Barreto et al. (2007) but within the range of 0.130 to 0.280mm stated by Genchev (2012).
With inclusion of autoclaved castor meal in the diets, a 10.2% relative increase in oil addition in these diets was necessary, which may be partially the cause of increased YC.However, the main influence may be the black color of castor seed coat in variety BRS Nordestina used in this experiment and possibly the presence of higher carotene concentration when compared with CSM diets.Sultana et al. (2007) stated that there may be effects on shell quality for different sources of calcium.Additionally, the main effect of SW and ESG reduction in eggs from quails subjected to ACM is assumed to be due to partial unavailability of calcium from CaO, which may have reacted with castor meal during autoclaving.Santos et al. (2015) evaluated a differently processed castor seed meal at the levels of 0, 5, 10, 15, and 20% and also found a decrease in EW and YW at the inclusion level of 20% when compared with CSM diet.However, in the present trial, Dunnett's test 1 (Table 3) revealed that values for ACMEC1 diet did not differ from those observed for CSM.The same effects detected for EW were also detected for YW.Thus, the use of EC did not provide differences when added to CSM diets, but comparing CSM diet with ACMEC1, EW and YW could be recovered, which was not the case with EC2.This effect can be the response of EC1 with supplementation of xylanase, β-glucanase, and phytase and lower concentration of cellulase, pectinase, and protease (secondary role), which acted by increasing the use of non-starchy polysaccharides contained in the autoclaved castor meal, generating a similar result for EW and YW.According to Babalola et al. (2006), β-xylanase addition in broiler diets containing boiled castor seed meal improved apparent nitrogen and fiber absorption as well as feed transit time.However, EC2 did not improve EW or YW in ACM diet, because it contains only amylase, xylanase and protease, probably having reduced the intensity of action on the fibers present in the autoclaved castor meal or even some indirect detrimental effect on endogenous enzyme production.
According to Olukosi et al. (2007), various enzyme complexes are used in animal feed to increase the bioavailability of nutrients.Lima et al. (2011) evaluated the effect of phytase in CSM diets for laying quails and concluded that production performance and egg quality were improved.However, in the present experiment, because of the formulation strategy ("over the top"), the effects of EC1 containing phytase in CSM diet were not observed.Bayram et al. (2008) evaluated the effect of adding xylanase in CSM diets for laying quails and also observed no differences in egg production or quality.Pernollet (1978), in turn, stated that castor bean meal has crystalloid and globoid protein body ultra-structures that are absent in soybean meal.In the autoclaved castor meal, the phytin (containing phytate) is located in globoids when present in protein bodies.The differences between

Table 1 -
Chemical composition of ingredients used in diets and calculated percentage composition of experimental diets according to diet type with or without autoclaved castor meal and addition of enzyme complexes.

Table 2 -
Composition and calculated analysis of experimental layer quail diets formulated with corn plus soybean meal or 21% autoclaved castor meal, with and without enzyme complex (EC).

Silva JCNS, Rabello CBV, Santos JS Laying Performance and Egg Quality of Japanese Quails Fed Diets Containing Castor Meal and Enzyme ComplexTable 3 -
Means (±standard deviation) for performance laying quails fed diets containing corn and soybean meal (CSM) or containing 21% autoclaved castor meal, with or without enzyme complex (EC), descriptive probability level for Dunnett's mean tests of significant parameters, and description of the variance analysis.

Table 4 -
Means (±standard deviation) for egg quality of quails fed diets containing corn and soybean meal (CSM) or containing 21% autoclaved castor meal, with or without enzyme complex (EC), descriptive probability level for Dunnett's mean tests of significant parameters, and description of the variance analysis.