Journal of Animal Science and Technology 53(5) 441~450, 2011 http://dx.doi.org/10.5187/jast.2011.53.5.441 농산부산물을이용한 In Vitro 반추위발효특성및적정배합수준을통한옥수수 및대두박대체효과 박중국 1 * 임동현 1 김상범 1 기광석 1 이현준 1 권응기 1 조원모 1 김창현 2 1, 2 Effects of Partial Replacement of Corn Grain and Soybean Meal with Agricultural By-Product Feeds on In Vitro Rumen Fermentation Characteristics and Optimum Levels of Mixing Ratio Joong Kook Park 1 *, Dong Hyun Lim 1, Sang Bum Kim 1, Kwang Seok Ki 1, Hyun June Lee 1, Eung Gi Kwon 1, Won Mo Cho 1 and Chang-Hyun Kim 2 1 National Institute of Animal Science, R.D.A., #9 Eoryong-ri, Seonghwan-eup, Cheonan-si, Chungcheongnam-do, 331-801, Rep. of Korea, 2 School of Animal Life and Environment Science, Hankyong National University, 327 Chungang-no, Anseong-si, Kyonggi-do, 456-749, Rep. Korea ABSTRACT This study was conducted to determine the effects of partial replacement of corn grain and soybean meal with agricultural by-product feeds on in vitro rumen fermentation characteristics and optimum levels of mixing ratio. The agricultural by-products to examine the effectiveness of the partial replacement of concentrate were wheat bran, corn gluten feed, bakery waste, soybean curd, rice bran, green kernel rice, soybean hull, distillers grain, and mushroom substrate. In the first experiment, in vitro ruminal fermentation characteristics of feedstuffs were evaluated at 0, 3, 6, 12, 24, and 48 hours after incubation. In the second experiment, fermentation characteristics were investigated with green kernel rice and soybean curd which replaced corn grain or soybean meal. Feed were formulated with 40% corn grain + 20% soybean meal (T1), 40% corn grain + 17.5% soybean meal + 2.5% soybean curd (T2), 25% corn grain + 20% soybean meal + 15% green kernel rice (T3), and 30% corn grain + 15% soybean meal + 6% green kernel rice + 9% soybean curd (T4), respectively, with forage source of 10% alfalfa hay, 20% timothy hay, and 10% corn silage as fed-basis. In 24 and 48 hour cultivations, T4 showed significantly lower ph compared to T1, whereas in 3 and 24 hour cultivations, T4 showed significantly higher DM degradation compared to T1. In addition, the gas production of T3 was also higher than T1 (p<0.05). Overall results of the present experiments indicated that green kernel rice and soybean curd as agricultural by-products have the possibility of partial replacements of corn grain and soybean meal. (Key words : Rumen, By-product, Concentrate, Soybean curd, Green kernel rice) 서론 60%,., 2009 3,954 22,599 24%, 30 (Ministry for Feed, Agriculture, Forestry and Fisheries, 2010)..,,,,, (Ministry for Food, Agriculture, Forestry and Fisheries, 2009b).,, (McDonnel, 1982; Farrell, 1994; Goh, 2002). * Corresponding author : Joong Kook Park, Dairy Science Division, National Institute of Animal Science, RDA, #9 Eoryong-ri, Seonghwan-eup, Cheonan-si, Chungcheongnam-do, 331-801, Rep. of Korea. Tel: +82-41-580-3399, Fax: +82-41-580-3419, E-mail: jkpark203@korea.kr - 441-
(Kwak and Yoon, 2003).,, (Ipharraguerre and Clark, 2003), (Vander Pol, 2008), (Ranathunga, 2010),. in vitro. 재료및방법 1., 9 (,,,,,,, ), Table 1. TMR (National Institute of Animal Science, 2009) TMR. 1 in vitro, 2., 2 in vitro, 2 in vitro (Table 2). 2,, (T1). T2 2.5%, T3 15%, T4 10 5% in vitro. 2. In vitro Holstein ( 650 kg) 1. 2, (08:00) (17:30) 5 kg 5 kg (Dactylis glommerata L.). 30, 4 2 (39 ) head space bottle. Table 1. Chemical composition of feedstuffs in the experiment Feedstuffs Items 1) DM CP EE CF CA NDF ADF NFC 2) % DM Corn grain 88.32 7.43 3.51 2.50 1.32 10.52 2.82 77.22 Soybean meal 90.33 45.30 1.95 6.00 7.01 14.03 8.09 31.71 Wheat bran 88.04 15.14 2.85 8.13 4.04 38.07 10.20 39.90 Corn gluten feed 92.64 19.33 1.35 9.83 8.26 35.38 11.69 35.68 Bakery waste 77.92 7.20 17.95 0.32 0.76 3.14 0.47 70.95 Soybean curd 13.44 22.30 4.34 15.60 3.47 33.86 19.45 36.03 Rice bran 87.14 13.62 19.47 7.37 9.19 23.82 10.78 33.90 Green kerneled rice 84.75 7.35 2.86 1.66 1.57 7.46 2.30 80.76 Soybean hull 91.85 11.43 1.24 29.69 5.97 56.01 38.08 25.35 Distillers grain 24.06 24.55 2.75 15.27 15.27 43.00 23.42 14.43 Mushroom substrate 25.50 10.49 0.92 27.27 11.61 58.47 38.60 18.51 1) DM: dry matter, EE: ether extract, CP: crude protein, CF: crude fiber, CA: crude ash, NDF: neutral detergent fiber, ADF: acid detergent fiber, NFC: non fiber carbohydrate. 2) NFC: nonfiber carbohydrates calculated as 100 (CP+EE+CA+NDF). - 442-
Table 2. Formulas and chemical composition of experimental diets T1 T2 T3 T4 Ingredient composition, % DM Corn grain 40.0 40.0 25.0 30.0 Soybean meal 20.0 17.5 20.0 15.0 Green kerneled rice 0.0 0.0 15.0 6.0 Soybean curd 0.0 2.5 0.0 9.0 Alfalfa hay 10.0 10.0 10.0 10.0 Timothy hay 20.0 20.0 20.0 20.0 Corn silage 10.0 10.0 10.0 10.0 Chemical composition, % DM Crude protein 15.94 15.37 15.93 15.38 Ether extract 2.42 2.48 2.33 2.54 Non fiber carbohydrate 1) 45.04 45.14 45.57 43.82 Neutral detergent fiber 29.02 29.51 28.56 30.74 Acid detergent fiber 18.89 18.15 17.81 19.05 Rumen degradable protein 2) 9.97 9.57 10.00 9.54 NE L 3), Mcal/d 15.76 15.85 15.71 15.80 1) Non fiber carbohydrates calculated as 100 (CP+EE+CA+NDF). 2),3) Rumen degradable protein and net energy for lactation were calculated according to NRC (2001).. Wiley mill 1 mm. 1 35 ml serum bottle 0.2 g 3, 2 0.4 g 3. blank serum bottle, 30 CO 2 bubbling ph 6.5 McDougall s buffer solution (Troelsen and Donna, 1966) 4 : 1 rumen inoculum. O 2 free CO 2. serum bottle. Serum bottle buffer 20 ml, 39 ± 0.5 (DS-10, Dasol Scientific Co., Korea) 3, 6, 12, 24 48. 3. (1) 가스발생량의측정 Williams (1996) Beuvink (1992) serum bottle 5 (burette). ml/g substrate DM. (2) 배양액의 ph 측정 serum bottle micro tube (3000 g) 15 ph meter (Gmbh8603, Mettler-Toledo, Switzerland) ph. (3) 배양액내암모니아태질소측정 Chaney Mabach (1962) 3, 6, 12, 24 48 serum bottle. (13,000 rpm) 5 NH 3 standard 0.02 ml 20 ml, blank tube. sample, standard blank phenol 50 g sodium nitroferricyanide [Na 2Fe (CN) 5NO 2H 2O] 0.25 g 1 L phenol color reagent 1 ml. 1 ml phenol color reagent - 443-
(NaOH) 25 g sodium hypochloride (4 6% NaCl) 16.8 ml 1 L alkalihypochlorite 1 ml. 37 15 8 ml. spectrophotometer 640 nm (optical density). (4) VFA 분석 VFA Erwin (1961). ph VFA microtubes (MCT-175-C, Axygen, USA) 1 ml 0.1 ml HgCl 2 25% HPO 3 0.2 ml 30. 20 13,000 rpm 10 4, gas chromatography (GC-14A, Shimadzu, Japan). (5) 건물분해율의측정 serum bottle. glass microanalysis filter holder assembly (No. 097531C, Fisher Scientific, USA) filter paper (Whatman No. 1, Whatman, UK) 80 drying oven 48. 4. (0, 3, 6, 12, 24 48 ) 3 SAS package (SAS Institute, Cary, NC) GLM (General Linear Model), Duncan s multiple range test (p<0.05). 결과및고찰...,. 2010 430 (Statistics Korea, 2010), (,, ) 6~7%.,.,. Farrell (1994), White Hembry (1985),.. TMR (Ministry for Food, Agriculture, Forestry and Fisheries, 2009a; Park and kim, 2010).,, (Vendramini and Arthington, 2009)., 15 20% (Chiou, 1998). NFC (77.22%) (80.76%), 70.95%. (45.30%) (24.55%), (22.30%), (19.33%) (15.14%) 15%. 1., NDF ADF 2. in vitro Table 1. (45.30%), (24.55%), (22.30), (80.76%), (77.22%), (70.95%)., 17.95%., ph Table 3 4., 6 ph, 24 48. 48,, ph, ph (p<0.05).,,, ph (p<0.05). Sievert Shaver - 444-
Table 3. Effects of feedstuffs on in vitro ruminal ph value in experiment 1 Incubation time CG GKR RB SH SC SEM 1) 0 hr 7.03 a 7.04 a 7.03 a 7.02 ab 7.00 b 0.005 3 hr 6.97 b 7.01 a 6.93 c 7.04 a 6.98 b 0.010 6 hr 6.89 c 6.93 b 6.87 d 6.97 a 6.92 b 0.009 12 hr 6.76 bc 6.78 b 6.74 c 6.92 a 6.66 d 0.023 24 hr 6.52 d 6.56 c 6.68 b 6.83 a 6.58 c 0.030 48 hr 6.28 e 6.34 d 6.61 b 6.67 a 6.39 c 0.041 Incubation time SM CGF WB DGS MS BW SEM 1) 0 hr 6.95 bc 6.93 c 7.00 a 6.99 a 6.99 a 6.97 ab 0.007 3 hr 6.93 b 6.90 c 6.87 d 6.92 bc 6.98 a 6.84 e 0.010 6 hr 6.88 b 6.88 b 6.81 c 6.89 b 6.97 a 6.66 d 0.024 12 hr 6.77 b 6.72 c 6.66 d 6.77 b 6.95 a 6.56 e 0.031 24 hr 6.74 b 6.56 d 6.56 d 6.69 c 6.87 a 6.37 e 0.039 48 hr 6.79 b 6.43 e 6.48 d 6.68 c 6.84 a 6.39 f 0.043 Abbreviated CG; corn grain, GKR; green kerneled rice, RB; rice bran, SH; soybean hull, SC; soybean curd, SM; soybean meal, CGF; corn gluten feed, WB; wheat bran, DGS; distillers grain with soluble, MS; mushroom substrate, BW; bakery waste. 1) SEM, standard error of means. a,b,c,d,e Means in the same row with different superscripts differ significantly (p<0.05). Table 4. Effects of feedstuffs on in vitro gas production (ml/g DM) in experiment 1 Incubation time CG GKR RB SH SC SEM 1) 3 hr 5.45 ab 4.50 cd 6.05 a 4.00 d 5.00 bc 0.292 6 hr 9.50 a 7.80 d 8.60 c 5.80 e 8.90 b 0.485 12 hr 14.90 b 14.80 b 11.65 c 9.00 d 19.50 a 1.274 24 hr 26.15 b 27.50 a 14.30 d 15.80 c 27.20 ab 2.430 48 hr 29.70 a 30.50 a 15.75 c 17.00 c 27.00 b 2.675 Incubation time SM CGF WB DGS MS BW SEM 1) 3 hr 6.50 b 6.77 b 8.50 a 5.43 c 4.00 d 8.03 a 0.372 6 hr 12.63 b 10.80 c 14.00 a 8.80 d 4.67 e 13.70 a 0.791 12 hr 19.37 c 17.63 d 21.47 a 14.40 e 7.90 f 20.10 b 1.109 24 hr 26.87 a 26.17 ab 26.83 a 18.20 c 13.53 d 25.77 b 1.255 48 hr 36.00 c 37.53 b 38.73 a 24.93 e 22.07 f 35.33 d 1.568 Abbreviated CG; corn grain, GKR; green kerneled rice, RB; rice bran, SH; soybean hull, SC; soybean curd, SM; soybean meal, CGF; corn gluten feed, WB; wheat bran, DGS; distillers grain with soluble, MS; mushroom substrate, BW; bakery waste. 1) SEM, standard error of means. a,b,c,d,e,f Means in the same row with different superscripts differ significantly (p<0.05). (1990) (NFC, non fiber carbohydrate) 34% 42% ph, NFC ph (Valadares, 1999). ph NFC,. In vitro, (Theodorou, 1994; 1998), (Beuvink, 1992). 1 12,,, 24, - 445-
(p<0.05)., (p<0.05). 1 12 48 (Table 5),. (Table 6). 6 Table 5. Effects of feedstuffs on in vitro DM degradation (%) in experiment 1 Incubation time CG GKR RB SH SC SEM 1) 0 hr 14.75 c 4.85 d 32.43 a 22.41 b 22.89 b 2.496 3 hr 10.55 c 13.07 c 38.85 a 24.93 b 25.18 b 2.724 6 hr 20.53 c 16.70 c 49.70 a 31.33 b 25.70 bc 3.478 12 hr 39.66 bc 34.61 c 55.00 a 41.64 b 50.78 a 2.172 24 hr 63.80 ab 67.31 a 56.49 b 54.49 b 70.34 a 2.140 48 hr 84.68 a 83.25 a 63.59 b 53.32 c 79.58 a 3.352 Incubation time SM CGF WB DGS MS BW SEM 1) 0 hr 28.92 bc 34.63 b 24.85 c 10.79 e 18.12 d 43.91 a 2.695 3 hr 43.95 a 34.84 b 32.77 b 26.23 b 27.92 b 45.30 a 2.096 6 hr 51.79 a 40.51 b 41.61 b 25.74 c 30.11 c 46.69 ab 2.432 12 hr 63.91 a 54.23 b 62.45 a 35.06 c 31.80 c 58.95 ab 3.205 24 hr 86.95 a 78.42 b 76.32 b 55.35 c 44.41 d 76.68 b 3.706 48 hr 96.77 a 91.55 b 86.78 c 61.76 d 52.85 e 84.03 c 3.914 Abbreviated CG; corn grain, GKR; green kerneled rice, RB; rice bran, SH; soybean hull, SC; soybean curd, SM; soybean meal, CGF; corn gluten feed, WB; wheat bran, DGS; distillers grain with soluble, MS; mushroom substrate, BW; bakery waste. 1) SEM, standard error of means. a,b,c,d Means in the same row with different superscripts differ significantly (p<0.05). Table 6. Effects of feedstuffs on in vitro ammonia-n concentration (mg/dl) in experiment 1 Incubation time CG GKR RB SH SC SEM 1) 0 hr 5.08 5.36 5.23 5.74 4.98 0.181 3 hr 5.39 d 6.84 cd 11.12 b 7.56 c 19.87 a 1.316 6 hr 1.37 d 2.50 cd 21.21 a 6.32 c 14.93 b 2.103 12 hr 1.40 c 1.45 c 13.63 a 4.92 b 11.77 a 1.415 24 hr 3.50 d 2.47 d 18.41 b 10.31 c 24.51 a 2.320 48 hr 15.61 d 16.74 d 32.60 b 22.42 c 50.82 a 3.516 Incubation time SM CGF WB DGS MS BW SEM 1) 0 hr 11.22 b 18.70 a 11.78 b 10.88 bc 6.34 d 8.20 cd 0.988 3 hr 13.66 c 9.12 d 24.54 b 16.94 c 30.00 a 8.87 d 2.051 6 hr 41.18 a 38.82 a 26.02 b 10.36 c 26.48 b 5.00 d 3.270 12 hr 57.07 a 28.31 b 28.21 b 17.71 c 29.26 b 12.00 c 3.522 24 hr 139.79 a 55.57 b 45.81 c 42.55 cd 35.89 d 14.74 e 9.646 48 hr 205.10 a 88.88 b 80.64 b 80.88 b 48.82 c 38.75 d 13.235 Abbreviated CG; corn grain, GKR; green kerneled rice, RB; rice bran, SH; soybean hull, SC; soybean curd, SM; soybean meal, CGF; corn gluten feed, WB; wheat bran, DGS; distillers grain with soluble, MS; mushroom substrate, BW; bakery waste. 1) SEM, standard error of means. a,b,c,d,e Means in the same row with different superscripts differ significantly (p<0.05). - 446-
(p<0.05),,., 7.35 7.43%. (Hristov, 1998; 2000). 3, 6 (p<0.05)., (p<0.05). in vitro, ph,,. 3. in vitro in vitro ph,, Fig. 1. ph 6.94 (0 h) 24 6.0. 12, 24 48 T4 (5.80 5.70) T1 (5.90 5.76) ph (p<0.05). 10 13% RDP (Hoover and Stokes, 1991) 35 42% NFC (Lykos, 1997),.,, ph 6.0 (Slyter, 1976). 6 T4, 24 48 T3 (p<0.05)., 12 12 24 T2 (2.62 18.80) T1 (4.91 21.09) (p<0.05). 48. (3 h) T1, (24 h) T4 (p<0.05). Fig. 1. Effects of substitution levels of concentrate with by-products on in vitro ph, gas production, ammonia-n, and DM degradation in experiment 2 Treatments are the same as describe in Table 2. - 447-
Table 7. Effects of substitution levels of concentrate with by-products on in vitro volatile fatty acid production (mm) in experiment 2 Incubation time Treatments 1) SEM 2) TI T2 T3 T4 Acetic acid 0 hr 30.90 33.07 33.10 32.93 0.443 3 hr 47.59 a 47.96 a 47.16 a 46.02 b 0.256 6 hr 52.03 c 53.36 bc 55.11 ab 56.39 a 0.559 12 hr 67.90 66.77 66.07 68.04 0.445 24 hr 78.34 ab 74.93 b 86.51 a 83.70 ab 1.898 48 hr 91.22 ab 88.01 b 92.71 a 92.14 a 0.722 Propionic acid 0 hr 6.78 7.40 7.29 7.11 0.109 3 hr 11.84 11.94 11.82 11.79 0.042 6 hr 13.87 c 14.39 bc 14.76 b 15.55 a 0.206 12 hr 19.89 a 19.39 bc 19.13 c 19.60 ab 0.103 24 hr 23.77 22.93 25.59 25.14 0.475 48 hr 27.74 27.85 27.97 28.28 0.152 Butyric acid 0 hr 4.25 b 4.59 a 4.61 a 4.40 ab 0.062 3 hr 6.69 6.68 6.56 6.45 0.047 6 hr 8.44 c 8.86 bc 9.11 ab 9.35 a 0.115 12 hr 14.72 a 14.06 b 14.17 b 13.45 c 0.142 24 hr 19.71 b 19.35 b 22.30 a 20.09 ab 0.464 48 hr 24.59 25.26 25.46 24.20 0.245 1) Treatments are the same as describe in Table 2. 2) SEM, standard error of means. a,b,c Means in the same row with different superscripts differ significantly (p<0.05). Table 7. Acetate 3 T4, T2 T3 T4 (p<0.05). acetate (Hutjens, 1998).,,., propionate butyrate 6 T1, 12 (p<0.05). 48.,. 요약 in vitro. 9 (,,,,,,,, ). 1 in vitro (0, 3, 6, 12, 24, 48 h), 2. 2 in vitro.,, - 448-
(T1). T2 2.5%, T3 15%, T4 10 5% in vitro. in vitro,. 2 ph, T4 T1, T3 T1 (p<0.05). 3 24 T4 T1 (p< 0.05).. ( 주제어 :,,,, ) 사사 2011. 인용문헌 Beuvink, J. M., Spoelstra, S. F. and Hogendorp, R. J. 1992. An automated method ofr measuring the time course of gas production of feedstuffs incubated with buffered rumen fluid. Neth. J. Agri. Sci. 40:401-407. Chaney, A. L. and Marbach, E. P. 1962. Modified reagents for determination of urea and ammonia. Clin. Biochem. 8:130-132. Chiou, P. W. S., Chen., C. R., Chen, K. J. and Yu, B. 1998. Wet brewer s grains or bean curd pomance as partial replacement of soybean meal for lactating cows. Anim. Feed Sci. Technol. 74:123-134. Erwin, E. S., Marco, S. J. and Emery, E. M. 1961. Volatile fatty acid analysis of blood and rumen fluid by gas chromatography. J. Dairy Sci. 44:1768-1771. Farrell, D. J. 1994. Utilization of rice bran in diets for domestic fowl and duckling. World s Poult. Sci. J. 50:115-131. Goh, B. D., Song, Y. H., Nakanishi, Y. and Manda, M. 2002. Effects of partial replacement of concentrate with soybean curd residue and Korean medical herb waste and feeding regime on nutrient digestibility and eating/resting behavior in sheep. Jpn. J. Livest. Management. 37:1342-1131. Hoover, W. H. and Stokes, S. R. 1991. Balancing carbohydrates and proteins for optimum rumen microbial yield. J. Dairy Sci. 74:3630-3644. Hristov, A. N., McAllister, T. A. and Cheng, K. J. 1998. Effect of dietary or abomasal supplementation of exogenous polysaccharide-degrading enzymes on rumen fermentation and nutrient digestibility. J. Anim. Sci. 76:3146-3156. Hristov, A. N., McAllister, T. A. and Cheng, K. J. 2000. Intraruminal supplementation with increasing levels of exogenous polysaccharide-degrading enzymes: effects on nutrient digestion in cattle fed a barley grain diet. J. Anim. Sci. 78:477-487. Hutjens, M. 1998. Feeding guide. Hoard's dairy man, W. D. Hoards & Sons Co. Ipharraguerre, I. R. and Clark, J. H. 2003. Soyhulls as an Alternative Feed for Lactating Dairy Cows: a review. J. Dairy Sci. 86:1052-1073. Kwak, W. S. and Yoon, J. S. 2003. On-site output survey and feed value evaluation in agro-industrial by-products. J. Anim. Sci. & Technol. (kor). 45:251-264. Lykos, T., Varga, G. A. and Casper, D. 1997. Varing degradation rates of total non-structural carbohydrates: Effects on ruminal fermentation, blood metabolotes, and milk production and composition in high producing Holstein cows. J. Dairy Sci. 80:3341-3355. McDonnell, M., Klopfenstein, T. and Merill, J. 1983. Soybean hulls can replace corn in growing rations. Beef cattle Report. University of Nebraska. MP. 44:17-18. Ministry for Food, Agriculture, Forestry and Fisheries. 2009a. Establishment of a feeding system and forage production technology for dairy goats in organic farming. pp. 65-71. Ministry for Food, Agriculture, Forestry and Fisheries. 2009b. Exploitation of organic feedstuffs in Korea and their data-base construction and development of both optimum organic feed processing and their feeding methods for organic animals. pp. 28-30. Ministry for Feed, Agriculture, Forestry and Fisheries. 2010. Statistics of Food, Agriculture, Forestry and Fisheries. pp. 362-364. National Institute of Animal Science, RDA. 2009. Handbook of TMR(total mixed ration) manufacturing plant in Korea. Park, J. K. and Kim, C. -H. 2010. Effects of organic feed containing rice bran and soybean hull on milk production of mid-lactation dairy goats. Kor. J. Organ. Agric. 18:599-612. Ranathunga, S. D., Kalscheur, K. F., Hippen, A. R. and Schingoethe, D. J. 2010. Replacement of starch from corn with nonforage fiber from distillers grains and soyhulls in diets of lactating dairy cows. J. Dairy Sci. 93:1086-1097. SAS. 2000. SAS/STAT Software for PC. SAS Institute Inc., Cary, NC, USA. Sievert, S. J. and Shaver, R. D. 1990. Effects of nonfiber carbohydrate level and Aspergillus oryzae fermentation extract on intake, milk production, and digestion in lactating dairy cows. J. Dairy Sci. 73(Suppl, 1):127.(Abstr). - 449-
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