44 4, (2017) Korean J. Poult. Sci. Vol.44, No.4, (2017) Effects of Dietary Si

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1 44 4, (2017) Korean J. Poult. Sci. Vol.44, No.4, (2017) Effects of Dietary Silicate Based Complex Mineral on Performance, Egg Quality and Immunological Competence in Laying Hens Chun Ik Lim 1, Jin Ern Park 1, Sang Eun Kim 2, Ho Sung Choe 3 and Kyeong Seon Ryu 1 1 Department of Animal Science, Chonbuk National University, Jeonju 54896, Republic of Korea 2 Davistone, Co., Ltd., Busan 48242, Republic of Korea 3 Department of Animal Biotechnology, Chonbuk National University, Jeonju 54896, Republic of Korea ABSTRACT This study was conducted to investigate the effect of feeding with dietary silicate based complex mineral (SCM) on the performance of laying hens. SCM at five levels (0%, 0.2%, 0.4%, 0.6% and 0.8%) was added to commercial diets, and fed to four hundred fifty Hy-Line Brown from fifty four to sixty five weeks of age. Egg production increased as the addition of dietary SCM to basal diets increased up to 0.6% (P<0.05). Furthermore, Feed intake increased in a manner similar to that of egg production (P<0.05). However, there was no difference in feed efficiency among the treatments. Eggshell thickness and breaking strength were significantly higher for the chickens fed with SCM than control from sixty weeks old to the end of experiment (P<0.05). Similarly, albumen height and Haugh unit were significantly higher for the chickens fed with SCM compared to those of control group (P<0.05). AST of birds fed SCM showed significantly higher than control (P<0.05). However, blood neutral fat level tended to increase in SCM treatments. Moreover, bone mineral density increased with SCM addition up to 0.4% (P<0.05). IL-2 (Interleukin-2) and IL-6 (Interleukin-6) levels appeared to be improved in the chickens fed with SCM addition diets, although not statistically different from all treatments. The results of this study indicated that the optimum SCM for improving the performance, egg quality and immunological competence of laying hens from 54 to 65 weeks of age was 0.6%. (Key words: egg quality, immunological competence, laying hens, performance, silicate complex mineral),, (zeolite), (bentonite), (kaolin) (biotite) (Safaeikatouli et al., 2011; Melegy et al., 2015; Yenice et al., 2015). (Kermanshahi et al., 2011), (Joo et al., 2007), (Melegy et al., 2015). (Mueller et al., 2004)., (Schwaller et al., 2016)., To whom correspondence should be addressed : seon@jbnu.ac.kr

2 268 :,. Table 2. Basal diet composition SiO 2 (73.2%) Al 2 O 3 (14.8%), CaO(1.37%) Fe 2 O 3 (0.61%),,,,. 54 Hy-Line 450 5, mm SiO 2 (73.2%) Al 2 O 3 (14.8%), CaO(1.37%) Fe 2 O 3 (0.61%) (Table 1) 0%, 0.2%, 0.4%, 0.6% 0.8%,, 2,800 kcal/kg, 16% (Table 2). 3, Table 1. SCM composition Mineral chemical composition (%) Ingredient % SiO Al 2 O Fe 2 O CaO MgO Na 2 O K 2 O Ignition loss Calcium Potassium Sodium Magnesium Iron Zinc Phosphorus Chemical composition Ingredient % Corn Soybean meal Corn gluten meal 4.24 Limestone 9.42 Calcium phosphate 0.94 Salt 0.38 L-Lysine 0.05 DL-Methionine 0.03 Vitamin premix Mineral premix Total ME (kcal/kg) 2,800 CP (%) Lysine (%) 0.74 Methionine (%) 0.32 Calcium (%) 3.80 Sodium (%) 0.17 Available phosphate (%) Contains per kg: vitamin A, 5,500 IU; vitamin D 3 1,100 ICU; vitamin E, 11 mg; vitamin B 12, mg; vitamin K 3, 1.1 mg; riboflavin, 4.4 mg; pantothenic acid, 11 mg (calcium pantothenate: 1.96 mg); choline, mg; folic acid, 0.55 mg; pyridoxine, 2.2 mg; biotin, 0.11 mg; thiamine, 2.2 mg; ethoxyquin, 125 mg. 2 Contains per kg; Cu, 10 mg; Fe, 60 mg; I, 0.46 mg; Mn, 120 mg; Zn, 100 mg., 16.,., 1.

3 Lim et al. : Effects of Dietary Silicate Based Complex Mineral on Performance, Egg Quality and Immunological Competence in Laying Hens 269,, (QC-SPA; TSS, UK), (FHK, Japan), (QCM+; TSS, England) g methanol benzen 4:1 5. acetyl chloride 200 μl heating block % potassium carbonate hexane 2 ml. (3,000 rpm, 4, 15 ) 20. Flame ionization detector (FID) 100 m 0.25 mm 0.2 μm, split ratio 30:1 GC-MSD (6890N-5973, Agilent, US). 10 (3000 rpm, 4, 10 ), (Automatic Biochemical Analyser, Thermo Scientitic, Konelab 20, Finland). 10, (Bone Densitometer, Pdexa, USA). RNA RNAiso plus(takara, Japan), RNA BioSpec-nano(Shimadzu, Kyoto, Japan). RNA 500ng 65 5, Rever- Tra Ace qpcr RT kit(toyobo, Japan) cdna, IL-2 IL-6, SYBR green PCR kit(topreal qpcr 2X Premix, enzynomics) CFX Connect Real-Time System(Bio-rad) PCR (Table 3). SAS(Statistical Analysis System, 9.2 Version, Cary, NC, 2002) General Linear Model(GLM), Duncan(1955) 0.05.,, 1, Table % 90.56% 87.74% (P<0.05), 1., 0.8% g (P<0.05).. Jeon et al.(2005) Kim et al.(2005),. Son(2005) 0.4%, Kermanshahi et al.(2011)., 0.6%. Table 3. Primer used for the quantitative real-time PCR Cytokines Primer sequence IL-2 IL-6 Forward 5 -GCTAATGACTACAGCTTATGGAGCA-3 Reverse 5 -TGGGTCTCAGTTGGTGTGTAGAG-3 Forward 5 -AAATCCCTCCTCGCCAATCT-3 Reverse 5 -CCCTCACGGTCTTCTCCATAAA-3

4 270 :, Table 4. Effect of feeding SCM on performance of laying hens Treatment (%) Egg production (%) Egg weight (g) Daily egg mass (g) Feed intake (g) Feed conversion c b bc b abc b a ab ab a SEM P value a c Value with the same letters in the row are significantly different at 5% level.,, Table %, 0.6% 0.8% 0.2% (P<0.05), 0.6% 3.75 kg/cm kg/cm kg/cm % (P< 0.05)., % (P<0.05). Fendri et al.(2012) 1.0% Table 5. Effect of feeding SCM on egg quality at 65 weeks of age Treatment (%) Shell breaking strength (kg/cm 2 ) Shell thickness (mm) Albumen height (mm) Haugh unit b b 7.97 c c ab b 8.23 bc bc a b 8.25 bc bc a a 8.82 ab ab a a 9.10 a a SEM P value 0.02 <0.01 <0.01 <0.01 a c Value with the same letters in the row are significantly di- fferent at 5% level.,. Mızrak et al.(2014) SiO 2 MgO 1.5 3%,. Yenice et al.(2015) 1.0% %. Table 6. Linoleic acid(c18:1 n-9), Palmitic acid(c16:0), Linoleic acid(c18:2 n- 6) Stearic acid(c18:0). Stearic acid(c18:0) Arachidonic acid(c20:4 n-6) 0.8 % (P< 0.05). Linoleic acid(c18:1 n-9) α-linolenic acid (C18:3 n-3) 0.8% % (P<0.05). (MUFA) (PUFA) (P<0.05), 0.8% 60.68% 64.29% (P<0.05). UFA/ SFA 0.8% (P<0.05) (P< 0.05). Son(2005) SiO 2

5 Lim et al. : Effects of Dietary Silicate Based Complex Mineral on Performance, Egg Quality and Immunological Competence in Laying Hens 271 Table 6. Effect of feeding SCM on fatty acid composition of egg yolk in laying hens at 65 weeks of age Treatment (%) C14:0 C16:0 C16:1 (n-7) C18:0 C18:1 (n-9) C18:2 (n-6) C18:3 (n-3) C20:1 (n-9) C20:4 (n-6) C22:6 (n-3) MUFA 1) PUFA 2) UFA 3) SFA 4) UFA/ SFA c a a 0.30 a 2.99 c a b a c 1.80 a b b a 0.27 b 3.66 b b ab b b 1.65 b bc b a 0.29 ab 3.43 b b ab bc ab 1.66 b b bc bc 0.27 b 3.83 b bc ab bc ab 1.59 bc a c c 0.26 b 4.55 a c a c a 1.54 c SEM P value <0.01 < < < <0.01 <0.01 <0.01 a c Value with the same letters in the row are significantly different at 5% level. 1) Monounsaturated fatty acid, 2) Polyunsaturated fatty acid, 3) Unsaturated fatty acid, 4) Saturated fatty acid. Fendri et al.(2012) 2%, MUFA PUFA. (Van, 1997).,, (Pond et al., 1988; Hagedorn et al., 1990; Kovar et al., 1990). Table 7., 0.8% ( mg/dl) ( mg/dl) 0.2% ( mg/dl) (P<0.05). AST, ALT, HDL-, % 2,294 2,358 mg/dl 0 0.4% 2,779 3,037 mg/dl. Lotfollahian et al.(2004) Safaeikatouli et al.(2011) Table 7. Effect of feeding SCM on blood composition of laying hens at 65 weeks of age Treatment (%) ALB 1) \ (g/dl) CHOL 2) (mg/dl) Glucose (mg/dl) AST 3) (IU/L) ALT 4) (IU/L) HDL 5) (mg/dl) Protein (g/dl) TG 6) (mg/dl) c , c , bc , ab , a , SEM P value a c Value with the same letters in the row are significantly different at 5% level. 1) Albumin, 2) Cholesterol, 3) Aspartate amino transferase, 4) Alanine amino transaminase, 5) High density lipoprotein cholesterol, 6) Trigly- cerides.

6 272 :,,, Miles and Henry(2007)., %, 15%.,. Fig % 0.2% (P<0.05). Leach et al.(1990) Elliot et al.(1990) zeolite,. (Schreiweis et al., 2003; Hester et al., 2004), 0.4%. IL-2 IL-6 (Janeway et al., 1999), IL-2 natural killer, B T γ, IL-8 IL-5, IL-6 B B. Fig. 2. IL-2 IL-6, 0.6%. Huff et al.(1988) Kubana and Harvey(1991) zeolite Hydrated sodium calcium aluminosilicate(hscas). Lee et al.(2009),, Joo et al.(2007) B., 0.6%. Fig. 1. Effect of feeding SCM on bone mineral density of laying hens at 65 weeks of age. Fig. 2. Effect of feeding SCM on expression of IL-2 and IL-6 in blood of laying hens at 65 weeks of age.

7 Lim et al. : Effects of Dietary Silicate Based Complex Mineral on Performance, Egg Quality and Immunological Competence in Laying Hens , 0%, 0.2%, 0.4%, 0.6% 0.8% ,,, 4,,. 0.6 % (P<0.05), (P<0.05),. (P<0.05), (P<0.05). (P<0.05), 0.4% (P<0.05). IL-2 IL-6 0.6% %. ( :,,,, ) Cutts JA, Wilson GC, Fernández S 2007 Optimum Egg Quality: A Practical Approach. 5M Publishing. Duncan DB 1955 Multiple range and multiple F tests. Biometrics 11:1-42. Elliot MA, Edwards BS, Burdette J 1990 Comparison of the effect of synthetic and natural zeolite on laying and broiler chickens performance. Poult Sci 70(10): Fendri I, Khannous L, Mallek Z, Traore AI, Gharsallah N, Gdoura R 2012 Influence of zeolite on fatty acid composition and egg quality in Tunisian Laying Hens. Lipids in Health and Disease 11(1):1-6. Hagedorn TK, Ingram DR, Kovar SJ, Achee VN, Barnes DG, Laurent SM 1990 Influence of sodium zeolite-a on performance, bone condition and liver lipid content of White Leghorn hens. Poult Sci 69(Suppl 1):169(Abstr). Han IK, Paik IK, Ha JK, Chae BJ 2012 Animal Nutrition. Seoul National University Press, pp Hester PY, Schreiweis MA, Orban JI, Mazzuco, H, Kopka MN, Ledur MC, Moody DE 2004 Assessing bone mineral density in vivo: dual energy X-ray absorptiometry. Poult Sci 83(2): Huff WE, Kubena LF, Harvey RBWE, Elissalde MH, Yersin AG, Philips TD Rottinghaus GF 1988 Toxic synergism between aflatoxin and T-2 toxin in broiler chickens. Poult Sci 67: Janeway CA, Traver P, Walport M, Shlomchik MJ 1999 Immunobiology: The Immune System in Health and Disease. Fourth Education Elsevier/Garland. London, pp Jeon HY, Son JH, Lee KW, Kim SK, Kang HS, Shin TS Cho BW 2005 Effects of feeding mixture of commercial broiler feed with spent bleaching clay from vegetable oil refinery on broiler performance. Korean J Poult Sci 32(4): Joo EJ, Jung SJ, Son JH, Cho JK, Yoon BS, Nam KT, Hwang SG 2007 Effect of dietary supplement of fermented clay mineral on the growth performance and immune stimulation in broiler chickens. Korean J Poult Sci 34(3): Kermanshahi H, Jani EHA, Hashemipour H, Pilevar M 2011 Efficacy of natural zeolite and pigments on yolk color and performance of laying hens. African J Biotechnol 10(16): Kim JH, Kim SC, Ko YD 2005 Effect of dietary zeolite treated on the performance and carcass characteristics in Fonishing pigs. J Anim Sci and Technol 47(4): Kovar SJ, Ingram DR, Hagedom TK, Achee, VN, Barnes DG, Laurent SM 1990 Broiler performance as influenced by sodium zeolite-a. Poult Sci 69(1):174. Kubena LF, Harvey RB 1991 Effect of hydrated sodium calcium aluminosilicate on growing turkey poults during aflatoxicosis. Poult Sci 70: Lee SR, Lee S, Chang KT, Kim JW 2009 Effects of dietary supplementation of illite on humoral immunity against Salmonella typhimurium flagella antigen in laying hens. Korean J Poult Sci 36(3): Leach RM, Heinrichs BS, Burdette J 1990 Broiler chicks fed low calcium diets. 1. Influence of zeolite on growth rate and parameters of bone metabolism. Poult Sci 69(9):1539-

8 274 :, Lotfollahian H, Shariatmadari F, Shivazad M, Mirhadi SA 2004 Study on the effects of two kinds of natural zeolite in diets on blood biochemical parameters, relative weight of body organs and broilers performance. Pajouhesh & Sazandegi 64: Melegy KME, Shehata SA, Abdel-Salam AF, Ragheb EM 2015 Influence of bentonite and ascorbic acid on minimizing the toxicity of aflatoxin B 1 in chickens diet. Egyptian J Poult Sci 35(2): Miles RD, Henry PR 2007 Safety of improved Milbond-TX when fed in broilers diets at greater than recommended levels. Anim. Feed Sci Technol 138: Mızrak C, Yenice E, Kahraman Z, Tunca M, Yıldırım U, Ceylan N 2014 Effects of dietary sepiolite and mannanoligosaccharide supplementation on the performance, egg quality, blood and digestion characteristics of laying hens receiving aflatoxin in their feed. Ankara University Vet Fak Derg 61: Mueller SO, Simon S, Chae K, Metzler M, Korach KS 2004 Phytoestrogen and their human metabolites show distinct agonistic properties on estrogen receptor alpha (ER alpha), and ER beta in human cell. Toxicol Sci 80(1): Pond WG, Yen JT, Varel VH 1988 Response of growing swine to dietary copper and clinoptilolite supplementation. Nutri Reports International 37(4): Safaeikatouli M, Jafariahangari Y, Baharlouei A 2011 An evaluation on the effects of dietary kaolin and zeolite on broilers blood parameters, T4, TSH and growth hormones. Pakistan J Nutri 10: SAS 2002 SAS/STAT Software for PC. SAS Institute Cary NC USA. Schreiweis MA, Orban JI, Ledur MC, Hester PY 2003 The use of densitometry to detect differences in bone mineral density and content of live White Leghorns fed varying levels of dietary calcium. Poult Sci 82(8): Schwaller D, Wilkens MR, Liesegang A 2016 Zeolite A effect on calcium homeostasis in growing goats. J Anim Sci 94 (4): Son JH 2005 Effects of dietary ceramic powder on laying performance, pathogenic bacterial counts in caecal contents and excreta, malodorous substances in excreta and fatty acid composition of egg yolk in laying hens. Korean J Poult Sci 32(4): Van Elswyk ME 1997 Comparison of n-3 fatty acid sources in laying hen rations for improvement of whole egg nutritional quality: A review. British J Nutri 78(1): Yenice E, Mizrak C, Ceylan N, Yildiz T, Gültekin M, Atik Z 2015 Effects of dietary sodium bentonite and mannan oligosaccharide supplementation on performance, egg quality, blood and digestion characteristics of laying hens fed aflatoxin contaminated diet. Kafkas University Vet Fak Derg 2: Received Oct. 18, 2017, Revised Dec. 17, 2017, Accepted Dec. 18, 2017

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