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40 1, 025 029 (2013) Korean J. Poult. Sci. Vol.40, No.1, 025 029 (2013) http://dx.doi.org/10.5536/kjps.2013.40.1.025 25 Assessment of Antimicrobial and Antioxidant Effects of Ripened Medicinal Herb Extracts to Select an Optimum Dietary Natural Antibiotic for Chickens Daekeun Shin, Ki-Woong Kim, Min-Jeong Jin, Kyeong Seon Ryu Department of Animal Science, Chonbuk National University, Jeonju 561-756, Korea ABSTRACT This study was conducted to evaluate the antimicrobial and antioxidant effects of ripened medicinal herb extracts from wood vinegar. The wood vinegar was collected from heated oaks. The Allium sativum (AS), Atractylodes ovate (AO), Cin- namomum zeylanicum (CZ), Coptidis rhizome (CR), Houttuynia cordata (HC), Phellodendron chinense (PC) and Syzygium aromaticum (SA) extracts were collected using wood vinegar, and they were ripened under room temperature for 50 days. All ripened medicinal herb extracts were used to test the antimicrobial and antioxidant activities. For the Lactobacillus, clear zone of 6 different medicinal herb except for CR ranged from 1.28 to 1.63 mm. 3.30 and 3.48 mm of clear zone were determined when CZ and SA were applied to Salmonella and E. coli, respectively (p<0.05), and they showed the largest clear zone as compared to other herbs. The clear zones of CR for Salmonella and E. coli were 2.21 and 3.34 mm, and each clear zone of CR was smaller than that of CZ (p<0.05) but was similar to that of SA (p>0.05). The amount of polyphenol and flavonoid was the highest in SA and CR, and they were 4.28 and 0.38 mg/ml, respectively (p<0.05). The CR shown 0.38 mg/ml flavonoid, had the highest DPPH, and it was 0.41 mm. The DPPH of HZ was significantly lowered in accordance with high amount of polyphenol and flavonoid, 2.56 and 0.20 mg/ml (p<0.05). In conclusion, CZ, SA, and CR showed high antimicrobial and antioxidant potentials, and therefore, may be used as alternatives to antibiotics for poultry diets. (Key words : antibiotics, antimicrobial, antioxidant, medicinal herb, poultry) 50,, (Guo et al., 2004; Yang et al., 2009)., 2003 53 (Kang et al., 2010).,, (Chae et al., 2011; Kim et al., 2011). 2011 7 (Park et al., 2012). Clostridium perfringens To whom correspondence should be addressed : seon@jbnu.ac.kr

26 :,, (Svobodova et al., 2007). (Dahiya et al., 2006).,. 200 (, 2006;, 2008). Salmonella typhimurium, Eschericha coli Lactobacillus rhamnosus., 1.2:1(w/w). (Table 1) 1:3(w/w), 26 50. 3,000 rpm 20. 7 100 ml 1 Eschericha coli (KCCM11234) Salmonella typhimurium (KCCM11862) Lactobacillus rhamnosus Kim et al. (2011). 10 cm plate 37 (MCO- Table 1. List of medicinal herbs used in this experiment Medicinal herb Scientific Name Korean name Manufacturer Allium sativum 마늘청운당농산, 경상남도함양군 Atractylodes ovate 백출휴먼허브, 경상남도경산시 Cinnamomum zeylanicum 계피경동정성재, 서울시 Coptidis rhizoma 황련휴먼허브, 경상남도경산시 Houttuynia cordata 어성초글로벌허브, 경상북도안동시 Phellodendron chinense 황백글로벌허브, 경상북도안동시 Syzygium aromaticum 정향휴먼허브, 경상남도경산시 15AC, Tokyo, Japan) 100 ul. 8 mm paper disc plate 4,, 50 ul paper disc 37 24 48 paper disc (clear zone) (mm)., 0.1 ml 1 ml 3, 1N Na 2 CO 3 90. 2 ml 725 nm (KR/Optizen 2,120 UV,,, ) (Juan and Chou, 2010). gallic acid. Juan and Chou (2010)., 25 ul 10 mm Tris HCl 1 ml 5% NaNO 3 75 ul, 6 10% AlCl 3 150 ul 5., 1M NaOH 500 ul EtOH 275 ul 510 nm (KR/Op- tizen 2,120 UV,,, ). Quercetin. 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) (Juan and Chou, 2010) DPPH 16 mg 100 ml 100 ml. 5 ml

Shin et al. : Medicinal Herb Antibiotic Effects 27, 1 ml 2, 528 nm (KR/Op- tizen 2,120 UV,,, ). 1 ml 5 ml,, (%). 전자공여능 시료의흡광도 대조구의흡광도 SAS(Version 9.2, Cary, NC, 2008), Duncan 95%. Table 2 7. 100 ml 1, (p>0.05). 2.56 mm, 6 1.28 1.63 mm (p<0.05). 3.30 mm,,,, 2.01 2.21 mm (p<0.05), (, 2006) (p>0.05)., (, 2008). (, 2002) 3.48 mm,,, (p>0.05). Table 2. Growth inhibiting activities of medicinal herbs extracts against different microbial strains Medicinal herb 1 Total plate count Growth inhibition zone (mm) Lactobacillus Salmonella E. coli Wood vinegar 1.93 ND 2 2.20 b 2.69 c Allium sativum 2.04 1.55 b 1.87 bc 3.26 ab Atractylodes ovate Cinnamomum zeylanicum 2.29 1.54 b 2.01 b 3.12 b 2.09 1.48 b 3.30 a 3.32 ab Coptidis rhizoma 2.29 2.56 a 2.21 b 3.34 ab Houttuynia cordata Phellodendron chinense Syzygium aromaticum 2.18 1.39 b 1.60 c 3.10 b 1.86 1.28 b 2.09 b 3.26 ab 2.30 1.63 b 2.18 b 3.48 a SEM 3 0.04 0.08 0.08 0.04 1 n = 15. 2 ND = not detected. 3 SEM = standard error mean. a c Means followed by different letter within the column are significantly different(p<0.05). 2.69 mm (p<0.05). 7, 4.28 mg/ml,,, 2.98 3.08 mg/ml (p<0.05)(table 3)., 2.56 2.60 mg/ml, 0.20 mg/ml (p<0.05). 0.15 mg/ml 0.38 0.22 mg/ml (p<0.05). DPPH(, 2008) 0.41 mm, eugenol thymol 0.26 mm (, 2002). DPPH, 0.18 mm DPPH (p<0.05).

28 : Table 3. Polyphenol and flavonoid contents and antioxidant activities of medicinal herbs extracts Medicinal herb 1 Antioxidant concentration 2 Polyphenol Flavonoid DPPH3 Wood vinegar 0.20 d - 0.21 e Allium sativum 2.59 c 0.19 bc 0.20 f Atractylodes ovate 2.99 b 0.19 bc 0.21 ef Cinnamomum zeylanicum 2.60 c 0.20 bc 0.24 c Coptidis rhizoma 3.08 b 0.38 a 0.41 a Houttuynia cordata 2.56 c 0.20 bc 0.18 g Phellodendron chinense Syzygium aromaticum 2.98 b 0.22 b 0.22 d 4.28 a 0.15 c 0.26 b SEM 4 0.22 0.03 0.01 1 n = 5. 2 Polyphenol = mg/ml; Flavonoid = mg/ml; DPPH = mm. 3 DPPH = 1,1-Diphenyl-2-picryl-hydrazyl. 4 SEM = standard error mean. a g Means followed by different letter within the column are significantly different(p<0.05).,.,,,,,, 50,. 6 1.28 1.63 mm, 3.30 3.48 mm. 2.21 mm 3.34 mm (p<0.05), (p>0.05). 4.28 0.38 mg/ml, 2.98 mg/kg 0.22 mg/kg. DPPH 0.38 mg/ml 0.41 mm, 2.56 mg/ml 0.20 mg/ml 0.18 mm DPPH (p<0.05).,,. ( :,,,, ). Chae MH, Nam HM, Jang GC, Kim HJ, Kim SR, Jung SC, Kang D, Kim, J 2011 Antimicrobial resistance in Campylobacter jejuni and Campylobacter coli isolated from food animals and raw meats in slaughterhouse in Korea during 2010. Kor J Vet Publ Hilth 35:239-245. Dahiya JP, Wilkie DC, Van Kessel AG, Drew MD 2006 Potential strategies for controlling necrotic enteritis in broiler chickens in post-antibiotic era. Anim Feed Sci Technol 129:60-88. Guo FC, Kwakkel RP, Soede J, Williams BA, Verstegen MWA 2004 Effect of a Chinese herb medicine formulation, as an alternative for antibiotics, on performance of broilers. Br Poult Sci 45:793-797. Juan MY, Chou CC 2010 Enhancement of antioxidant activity, total phenolic and flavonoid content of black soybeans by solid state fermentation with Bacillus subtilis BCRC 14715. Food Microbiol 27:586-591. Kang GH, Kim SH, Kim JH, Kang HK, Kim DW, Cho SH, Seong PN, Park BY, Kim DH 2010 Effects of environmental temperature and antibiotic substitute on quality of chicken breast meat. Korean J Food Sci Ani Resour 30:261-268. Kim SR, Nam HM, Jang GC, Kim AR, Kang MS, Chae MH, Jung SC, Kang D, Kim JK 2011 Antimicrobial resistance in Campylobacter jejuni and Campylobacter coli isolated

Shin et al. : Medicinal Herb Antibiotic Effects 29 from food animas and raw meats in slaughterhouse in Korea during 2010. Kor J Vet Publ Hlth 35:246-254. Park YS, Kim JR, Kang BK, Choe YH, Kim BS 2012 Inhibitory effects of herbal medicine on coccidiosis. Korean J Vet Serv 35:25-31. SAS 2008 SAS/STAT Software. Release 9.2, SAS Inst Inc Cary, NC, USA. Svobodova I, Steinhauserova I, Nebola M 2007 Incidence of Clostridium perfringens in broiler chickens in the Czech Republic. Acta Vet Brno 76:S25-S30. Yang Y, IJI PA, Choct M 2009 Dietary modulation of gut microflora in broiler chickens: a review of the role of six kinds of alternatives to in-feed antibiotics. World Poult Sci J 65:97-114. 2006. 21:145-152. 2008 ethanol. 27: 156-162. 2002. 12: 431-453. 2006. 33:283-293. 2008,, (Albac G150). 35:29-37. ( : 2013. 1. 31, : 2013. 3. 14, : 2013. 3. 15)