International Journal of Poultry Science, 8 (1): 80-89, 2009ISSN 1682-8356 Asian Network for Scientific Information, 2009

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Effects of Copper Sulfate on Productive, Reproductive Performance and BloodConstituents of Laying Japanese Quail Fed Optimal and Sub-Optimal Protein

I.M. Abaza, W. Ezzat, M.S. Shoeib, A.A. El- Zaiat and I.I. HassanAnimal Production Research Institute, Agriculture Research Center, Dokki, Giza, Egypt

Abstract: A (3X3) factorial design experiment was conducted to study the effect of three levels of crudeprotein (16, 18 and 20%) and three levels of copper sulfate (0, 100 and 200 mg/kg diet) as a growth promoteron productive and reproductive performances, egg quality, blood serum constituents and economicalefficiency of laying quail hens through 8 weeks. A total number of 270 hens and 135 males of Japanese quailat 8 weeks of age with nearly equal body weight and average rate of laying were randomly divided into 9groups (30 hens and 15 males each). Each group of birds was sub divided into 3 replicates (10 hens and5 males) and each replicate was housed in one wire cage. The results showed that the layer body weightsat 12 or 16 weeks and weight gain at 12-16 and 8-16 weeks of age were significantly increased withincreasing crude protein level from 16-18 or 20%, while there were no significant differences between thegroups fed 18 and 20% crude protein throughout the experimental intervals and the whole period. Eggnumber, rate of laying and egg mass of laying quail hens increased with increasing crude protein at levels16-18 or 20% (except at 12-16 week of age), while insignificant differences were found between the groupsfed 18 and 20% crude protein throughout the experimental intervals and the whole period. Addition of coppersulfate at levels of 100 and 200 mg/kg to laying quail diets significantly improved egg number, rate of laying,egg mass and feed conversion ratio except at 8-12 weeks of age as compared with group non-supplemented with copper sulfate during the experimental period. The highest values of body weight andegg mass were recorded with 20 % protein plus 100 mg copper sulfate/kg diet, while, the best values of feedconversion and The highest values of egg number and rate of laying were recorded with 20 % protein plus200 mg copper sulfate/kg diet from 8-16 weeks of age for quail layer as compared with other treatments.Laying quails hens fed diet contained copper sulfate levels significantly increased hatchability of fertile eggspercentage (except at 12 wks of age) as compared with those un-supplemented group. Various levels ofcrude protein or copper sulfate containing diets did not significantly affect on egg quality parameters, whileegg yolk cholesterol was significantly decreased with increased copper sulfate levels. The highest valuesfor total serum protein and serum albumin were recorded with experimental groups fed 18 or 20% proteinlevels compared with those received 16% protein diet, while serum cholesterol values were significantlydecreased with increased crude protein. However, dietary copper sulfate supplementation reduced serumtri-glycerides, total cholesterol and low density lipoprotein cholesterol, but increased serum high densitylipoprotein cholesterol. The best value of economic efficiency was recorded with laying quail hens fed 18%protein with 100 mg copper sulfate/kg diet compared with other treatment groups.

Key words: Protein, copper, feed supplement, performance, quails hens

INTRODUCTIONRecently, feeding cost in poultry production isconsidered the most expensive item (accounts for 65-70% of the total costs) particularly, dietary protein whichis the most expensive in poultry diets. High-protein diets(20% crude protein CP) are recommended for Japanesequail layers for optimizing performance (NRC, 1994). (El-Hammady et al.,1992) found that, feeding low proteindiets had no adverse effects on laying hens comparedto high protein diets. Also (Abdel-Azeem et al., 2005)found that when dietary protein level reduced up to 16 %,the performance values of laying quail hens weredecreased.Copper sulfate is a naturally-occurring inorganic salt andcopper is an essential trace element in poultry nutrition

(Davis and Mertz, 1987), copper deficiency in laying hensresulted in hypercholesterolemia (Kelvay et al., 1984),proper bone growth and development as well as enzymefunctions. Copper is often added to poultry diets atprophylactic concentrations for its growth promotingeffects (Pesti and Bakalli, 1996). Traditionally, the sourceof copper has been copper sulfate pentahydrate due tocost and commercial availability. In recent years,additional copper sources have become available andthe potential for commercial use as feed supplementhas expanded. Copper is usually fed commercially atmuch higher pharmacological levels (100-300 mg/kgdiet) because of its growth promoting properties (Bakalliet al., 1995). (Pearce et al.,1983) demonstrated thatpharmacological levels of Cu (>250 mg/kg diet) caused

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changes in 17 beta-estradiol and enzymes involved incarbohydrate, lipid and amino acid metabolism inmature laying hens and suggested that coppersupplements can affect reproductive physiology and lipidmetabolism. Egg production and feed conversion oflaying hens were improved by addition of 300 mg Cu/kgof diet (Metwally, 2002). (Idowu et al., 2006) clear thatincreasing the supplementation of copper sulfatepentahydrate, CuSO . 5H O and Copper Proteinate4 2(CuP) from 0-250 mg Cu/kg of laying hens dietincreased egg production and Haugh unite while thefeed intake was decreased. (Arias and Koutsos, 2006)demonstrate that broiler performance and intestinalphysiology can be positive effected by dietary coppersource at level of 188 mg Cu/kg diet as well as microbialenvironment. Addition of different amounts of copper tolaying hens diet have resulted in reductions in yolkcholesterol concentrations and blood plasma lipidconcentrations (Pesti and Bakalli, 1998). Therefore, this study was conducted to evaluate theeffect of dietary crude protein levels and copper sulfatelevels as a growth promoter on productive andreproductive performance, egg quality, bloodconstituents and economical efficiency of laying quailhens.

MATERIALS AND METHODSThe present study was carried out at the PoultryResearch Farm of Poultry Production, Faculty ofAgriculture, Zagazig University, Egypt. A (3X3) factorialdesign experiment was conducted to study the effect ofthree levels of crude protein CP (16, 18 and 20%) andthree levels of copper sulfate CuSO (0, 100 and 200 mg4/kg diet) on some productive and reproductiveperformance, egg quality, serum blood constituents andeconomical efficiency of Japanese quail layers. Dietswere formulated to contain 16, 18 or 20% Crude Protein(CP) supplemented with methionine and lysine to meetthe requirements recommended by (NRC, 1994) and2670 k cal ME/kg (Table 1).The ninth experimental treatments were as follows:Diet 1) 16% CP + 0 mg CuSO / kg diet.4Diet 2) 16% CP + 100 mg CuSO / kg diet.4Diet 3) 16% CP + 200 mg CuSO / kg diet.4Diet 4) 18% CP + 0 mg CuSO / kg diet.4Diet 5) 18% CP + 100 mg CuSO / kg diet.4Diet 6) 18% CP + 200 mg CuSO / kg diet.4Diet 7) 20% CP + 0 mg CuSO / kg diet.4Diet 8) 20% CP + 100 mg CuSO / kg diet.4Diet 9) 20% CP + 200 mg CuSO / kg diet.4

A total number of 270 hens and 135 males of Japanesequail at 8 weeks of age with nearly equal body weightand average rate of laying were randomly divided into 9treatment groups (30 hens and 15 males each). Eachgroup of birds was sub divided into 3 replicates (10

Table 1: Composition and calculated analysis of experimentalcorn-soy diets fed during the laying quail period (8-16wks of age)

Crude Protein level--------------------------------------------------

Ingredients 16% 18% 20%Yellow corn 62.24 59.80 55.86Soybean meal 20.16 27.16 33.41Wheat bran 9.32 5.00 3.00Limestone (CaCO ) 5.05 5.05 5.253Dicalcium phosphate 2.25 2.20 1.80Vit. & Min. Premix* 0.30 0.30 0.30Sodium chloride 0.25 0.25 0.25DL-methionine 0.19 0.16 0.13Lysine 0.24 0.08 0Total 100 100 100Chemical analysis:Calculated analysis **ME Kcal/kg 2688 2683 2660Crude protein %*** 15.91 17.93 19.85Ether extract (EE) *** 2.61 2.44 2.25Crude fiber %*** 3.68 3.63 3.77Calcium % 2.50 2.50 2.50Available phosphorus % 0.53 0.52 0.47Lysine % 1.00 1.00 1.05Methionine % 0.45 0.45 0.45Copper (mg) 10.05 11.17 12.27*: Each 3 kg of Vit. And Min. premix contains 10000000 IU Vit. A;2000000 IU Vit. D3; 10000 mg Vit. E; 1000 mg Vit. K3; 1000 mgVit. B1; 5000 mg Vit. B2; 10 mg Vit B12; 1500 mg Vit. B6; 30000mg Niacin; 10000 mg Pantothenic acid; 1000 mg Folic acid; 50mg Biotin; 300000 mg Choline chloride; 4000 mg Copper; 300mg Iodine; 30000 mg Iron; 50000 mg Zinc; 60000 mgManganese; 100 mg Selenium and 00 mg Cobalt. **Calculatedaccording to (NRC, 1994). ***Determined according to themethods of A.O.A.C (1990).

hens and 5 males each) and each replicate was housedin one wire cage. Birds were fed ad libitum and the freshwater was available all time during the experimentalperiod. Artificial light was used giving a total of 16 h oflight per day throughout the experimental period.Individual body weights were recorded at (8, 12 and 16weeks of age) and body weight gain was calculated forintervals (8-12, 12-16 and 8-16 weeks of age). Theproductive performance of layers was estimated at (8-12, 12-16 and 8-16 weeks of age) For each replicate eggnumber and egg weight were recorded daily. Rate oflaying (egg/hen/day), feed intake and egg mass werecalculated weekly. Feed conversion was calculated (gfeed intake/g egg mass/hen). At the 12 and 16 weeks of age, 135 eggs from eachtreatment were collected and incubated till hatching.Percentages of fertility and hatchability of total fertileeggs were estimated. Egg quality traits were determinedat 16 weeks of age on 5 eggs from each replicate. Eggshape index, yolk index, yolk, albumen and shellpercentages, also haugh units score were calculated. At the end of experimental period, blood samples werewithdrawn from three birds in each group from Jugular

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vein in tubes. Blood was centrifuged for 20 min at 4000 with increasing protein level in the laying quail hens diet.g and blood serum samples were stored at -20 C untilo

analysis of the total protein, albumin, total cholesterol,high density lipoprotein cholesterol, low densitylipoprotein cholesterol, triglycerides and glucose. Theanalyses of crude protein in the diet were carried outaccording to A.O.A.C. (1990). Cholesterol was extractedfrom yolk samples by chloroform method (2:1) accordingto (Folch et al., 1957). Total protein, albumin, totalcholesterol, High Density Lipoprotein Cholesterol(HDL%), Low Density Lipoprotein Cholesterol (LDL%),triglycerides and glucose concentrations weredetermined in the serum. Yolk cholesterol byspectrophotometer method using commercial kits(Stanbio Laboratory, Boerne, Texas 78006, USA).Economical efficiency for egg production was calculatedfrom the input/output analysis according to the price ofexperimental diets and eggs production. Values ofeconomical efficiency were calculated as the netrevenue per unit of total costs (Osman, 2003 andSoliman et al., 2003). Data were subject to factorialdesign using general linear model of SAS softwarestatistical analysis (SAS, 1998). The Significant meanswere separated by Duncan's New Multiple Range test(Duncan, 1955).

RESULTS AND DISCUSSIONProductive performance: The data in Table 2 showsthat layers fed CP at level of (20%) had the highestaverage of body weight and weight gain at both (12 and16 wks of age) and through (12-16 and 8-16 wks of age)intervals. While those fed CP at level of (16%) had thelowest average of these traits while layers fed (18%) hadintermediate values. The CP did not affective either bodyweight or weight gain at the early period of growth.These results are in agreement with the findings of(Soares et al., 2003) who observed that laying wasdelayed and varied in body weight for quails fed lowerprotein level (16% CP). Also (Ali et al., 2000) found that,high protein level in the quail diets, resulted in higherbody weight and weight gain than the low protein leveldiet. (Abdel-Azeem et al., 2005) observed that the significantimprovements in live body weight and weight gain forquail hens received high protein diets (20% CP).The data in Tables 3 and 4 shows that egg number, rateof laying and egg mass were significantly (p

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Table 2: Means SE of body Weight and body Weight gain of laying Japanese quail as affected by dietary crude protein levels withor without copper sulfate supplementation during the experimental periods

Body Weight (g) Weight gain (g)--------------------------------------------------------------------- ------------------------------------------------------------------------

Items 8 wks 12 wks 16 wks 8-12 wks 12-16 wks 8-16 wksProtein levels (CP)16% 207.310.89 223.740.54 243.531.14 16.431.08 19.790.95 36.221.27b b b b

18% 209.520.51 225.270.47 247.340.99 15.750.63 22.071.19 37.821.12ab a ab ab

20% 209.141.05 226.560.95 250.391.21 17.420.56 23.830.57 41.250.83a a a a

Significant NS * * NS * *CuSO levels (mg/kg)40 208.360.87 225.160.48 245.881.50 16.800.72 20.721.09 37.521.26100 209.220.96 226.270.88 248.761.43 17.050.69 22.480.76 39.540.78200 208.390.87 224.140.78 246.631.35 15.750.97 22.491.28 38.241.66Significant NS NS NS NS NS NSInteraction effects (CP X CuSO levels)416% CPX0 CuSO 206.652.35 223.850.76 241.612.07 17.201.61 17.771.42 34.961.334 b c

16% CPX100 CuSO 206.570.58 223.671.28 244.321.05 17.101.80 20.651.44 37.751.514 b bc

16% CPX200 CuSO 208.701.46 223.701.14 244.662.72 15.002.55 20.961.94 35.963.664 b bc

18% CPX0 CuSO 208.960.47 225.190.45 245.710.95 16.230.91 20.521.04 36.751.174 b bc

18% CPX100 CuSO 209.571.44 226.170.47 248.540.87 16.601.12 22.380.63 38.970.584 ab ab

18% CPX200 CuSO 210.030.70 224.451.24 247.772.80 14.421.18 23.323.65 37.753.494 b ab

20% CPX0 CuSO 209.470.99 226.440.60 250.311.73 16.971.59 23.871.32 40.832.634 ab ab

20% CPX100 CuSO 211.521.58 228.990.62 253.411.43 17.471.00 24.420.98 41.890.504 a a

20% CPX200 CuSO 206.441.80 224.262.10 247.451.89 17.820.31 23.190.90 41.020.884 b abc

Significant NS * * NS NS NSMeans having different letters at the same column are differ significantly. * = (p

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Table 3: Means SE of egg number and rate of laying of laying Japanese quail as affected by dietary crude protein levels with orwithout copper sulfate supplementation during the experimental periods

Egg number (egg/hen) Rate of laying (egg/hen/day) %------------------------------------------------------------------- -----------------------------------------------------------------

Items 8-12 wks 12-16 wks 8-16 wks 8-12 wks 12-16 wks 8-16 wksProtein levels (CP)16% 19.330.23 22.560.34 41.320.60 69.050.84 80.551.22 74.800.86b b b b

18% 20.310.38 23.000.32 43.540.68 72.541.36 82.141.14 77.341.06ab a ab ab

20% 20.620.40 23.460.35 44.120.57 73.651.44 83.771.26 78.711.00a a a a

Significant * NS * * NS *CuSO levels (mg/kg)40 19.600.29 22.220.33 41.380.57 70.001.03 79.371.17 74.680.78b b b b

100 20.170.47 23.340.34 43.540.71 72.021.66 83.371.20 77.701.28a a a a

200 20.500.35 23.440.34 44.070.59 73.211.25 83.730.87 78.470.79a a a a

Significant NS * * NS * *Interaction effects (CP X CuSO levels):-416% CPX0 CuSO 19.030.26 22.100.61 39.800.75 67.980.93 78.932.18 73.451.244 d c

16% CPX100 CuSO 19.130.52 22.630.71 41.771.23 68.341.87 80.832.54 74.582.204 bcd bc

16% CPX200 CuSO 19.830.35 22.930.59 42.400.64 70.831.26 81.912.10 76.370.624 abcd abc

18% CPX0 CuSO 19.330.29 22.030.62 41.370.88 69.051.04 78.692.22 73.871.584 cd c

18% CPX100 Cu so 20.730.73 23.530.34 44.370.35 74.052.61 84.051.21 79.050.774 ab ab

18% CPX200 Cu so 20.870.62 23.430.15 44.901.01 74.532.22 83.690.52 79.111.154 a ab

20% CPX0 Cu so 20.430.55 22.530.67 42.970.13 72.981.96 80.482.39 76.720.244 abc abc

20% CPX100 Cu so 20.630.99 23.870.58 44.501.45 73.693.55 85.242.06 79.472.594 ab ab

20% CPX200 CuSO 20.800.79 23.970.27 44.900.83 74.282.83 85.600.97 79.941.514 a a

Significant NS NS * NS NS *Means having different letters at the same column are differ significantly. * = (P

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Table 4: Means SE of egg weight and egg mass of laying Japanese quail as affected by dietary crude protein levels with or withoutcopper sulfate supplementation during the experimental periods

Egg weight (g) Egg mass (g/hen) ------------------------------------------------------------------ -----------------------------------------------------------------------

Items 8-12 wks 12-16 wks 8-16 wks 8-12 wks 12-16 wks 8-16 wksProtein levels (CP)16% 10.880.03 10.880.02 10.880.02 210.272.49 245.493.79 455.765.24b b

18% 10.880.02 10.790.10 10.830.06 220.974.17 248.043.78 469.016.60ab ab

20% 10.870.04 10.900.02 10.890.03 224.124.02 255.603.78 479.725.61a a

Significant NS NS NS * NS *CuSO levels (mg/kg)40 10.920.02 10.890.01 10.910.01 214.033.11 242.093.73 456.124.85b b

100 10.860.02 10.920.01 10.890.01 219.105.21 254.883.80 473.98.94a a

200 10.840.04 10.760.10 10.800.06 222.233.43 252.153.18 474.384.97ab a

Significant NS NS NS NS * *Interaction effects (CP X CuSO levels)416% CPX0 CuSO 10.960.04 10.880.04 10.920.02 208.553.16 240.397.04 448.947.03416% CPX100 CuSO 10.820.05 10.920.02 10.870.03 206.986.04 247.258.02 454.2214.06416% CPX200 CuSO 10.860.06 10.850.02 10.860.03 215.292.88 248.826.23 464.114.77418% CP X 0 CuSO 10.880.02 10.890.03 10.880.01 210.272.81 239.917.47 450.1810.10418% CPX100 CuSO 10.890.03 10.900.02 10.890.02 225.748.23 256.454.17 482.184.27418% CPX200 CuSO 10.870.05 10.580.29 10.730.17 226.896.89 247.775.43 474.6611.08420% CPX0 CuSO 10.930.04 10.920.01 10.920.02 223.265.80 245.987.10 469.231.36420% CPX100 CuSO 10.890.03 10.930.01 10.910.02 224.6010.56 260.956.52 485.5515.66420% CPX200 CuSO 10.800.10 10.840.05 10.820.07 224.516.91 259.872.90 484.386.944Significant NS NS NS NS NS NSMeans having different letters at the same column are differ significantly. * = (p

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Table 6: Means SE of reproductive performance of laying Japanese quail as affected by dietary crude protein levels with or withoutcopper sulfate supplementation at 12 and 16 wks of age

Fertility % Hatchability of total eggs % Hatchability of fertile eggs%----------------------------------------- ------------------------------------------ --------------------------------------------------

Items 12 wks 16 wks 12 wks 16 wks 12 wks 16 wksProtein levels (CP)16% 82.871.93 79.350.48 68.041.87 65.370.33 82.071.00 82.410.60b

18% 83.130.86 80.901.12 68.570.94 67.011.02 82.490.83 82.840.86ab

20% 85.550.98 82.590.89 70.621.36 68.030.57 82.510.91 82.420.78a

Significant NS NS NS * NS NSCuSO levels (mg/kg)40 82.301.18 81.160.99 66.641.38 65.900.84 80.971.22 81.190.50b

100 84.171.15 80.811.06 69.811.08 67.760.67 82.950.74 83.900.86a

200 85.091.66 80.860.90 70.781.57 66.760.74 83.150.33 82.580.53ab

Significant NS NS NS NS NS *Interaction effects (CP X CuSO levels)416% CPX0 CuSO 81.263.77 79.951.37 65.714.13 64.640.45 80.782.35 80.881.00416% CPX100 CuSO 84.642.53 78.650.33 69.741.82 66.160.26 82.462.16 84.120.18416% CP X 200 CuSO 82.714.60 79.460.54 68.674.16 65.320.68 82.980.59 82.220.84418% CPX0 CuSO 82.191.13 79.901.59 67.261.44 64.460.52 81.902.61 80.721.15418% CPX100 CuSO 82.431.68 81.562.48 68.061.71 68.711.76 82.560.99 84.301.61418% CPX200 CuSO 84.791.66 81.252.37 70.381.73 67.842.00 83.000.42 83.521.22420% CPX0 CuSO 83.440.30 83.651.67 66.931.69 68.591.59 80.232.21 81.980.52420% CPX100 CuSO 85.452.06 82.231.88 71.652.10 68.400.51 83.830.58 83.292.45420% CPX200 CuSO 87.761.60 81.881.53 73.272.00 67.120.70 83.460.85 82.000.684Significant NS NS NS NS NS NSMeans having different letters at the same column are differ significantly. * = (p

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Table 8: Means SE of serum biochemical traits for laying Japanese quail as affected by dietary crude protein levels with or without copper sulfatesupplementation at the end of the experimental period

Total protein Albumin Globulin Tri-glycerides Cholesterol GlucoseItems (mg/100 mL) (mg/100 mL) (mg/100 mL) (mg/dl) (mg/dl) HDL% LDL% (mg/100 mL)Protein levels (CP)16% 5.180.15 2.160.06 3.020.14 204.932.04 167.841.73 30.980.84 69.020.84 86.930.88b b a

18% 5.870.13 2.640.10 3.220.20 202.962.44 163.282.24 31.860.93 68.140.93 86.571.18a a ab

20% 6.050.16 2.830.07 3.210.18 205.171.93 160.422.30 31.570.99 68.430.99 87.201.31a a b

Significant ** ** NS NS * NS NS NSCuSO levels (mg/kg)40 5.500.21 2.520.12 2.980.12 210.100.88 168.621.59 29.320.61 70.680.61 87.350.98 a a b a

100 5.770.20 2.570.14 3.200.19 201.662.24 162.891.83 32.520.70 67.480.70 86.291.12 b b a b

200 5.820.16 2.550.12 3.270.19 201.291.48 160.032.47 32.570.90 67.430.93 87.061.26 b b a b

Significant NS NS NS ** * * * NSInteraction effects(CP X CuSO levels):416% CPX0 CuSO 4.900.34 2.160.15 2.740.26 209.242.13 170.702.07 28.990.40 71.010.40 87.041.964 b b ab a

16% CPX100 CuSO 5.240.24 2.160.13 3.080.14 201.114.62 165.243.58 32.321.85 67.681.85 86.231.744 ab b abc abc

16% CPX200 CuSO 5.390.22 2.170.08 3.230.30 204.442.68 167.583.38 31.631.37 68.371.37 87.521.444 ab b abc abc

18% CPX0 CuSO 5.610.13 2.610.18 3.000.10 210.171.69 168.753.77 29.000.82 71.000.82 87.231.984 ab ab a ab

18% CPX100 CuSO 5.960.28 2.690.20 3.270.48 199.075.24 163.283.41 32.81 1.00 67.191.00 85.872.104 a a c abcd

18% CPX200 CuSO 6.030.23 2.640.19 3.400.42 199.631.47 157.812.07 33.781.55 66.221.55 86.602.804 a ab bc cd

20% CPX0 CuSO 5.980.30 2.790.12 3.190.23 210.911.12 166.412.70 29.981.80 70.021.80 87.771.934 a a a abc

20% CPX100 CuSO 6.120.36 2.870.17 3.250.43 204.801.76 160.163.05 32.441.22 67.561.22 86.762.754 a a abc bcd

20% CPX200 CuSO 6.030.27 2.840.14 3.190.36 199.813.11 154.693.52 32.302.24 67.702.24 87.073.014 a a bc d

Significant * * NS * * NS NS NSMeans having different letters at the same column are differ significantly. * = (p

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