인진쑥 및 녹차 추출물을 이용한 무항생제 닭고기 생산 연구 = Studies for Antibiotic Free Chicken Production Using Water Extracts from Artemisia capillaris and Camellia sinensis

국문 초록 (Abstract)

본 연구는 무항생제 닭고기 생산을 위한 성장촉진용 항생제를 대체할 천연생리활성물질 개발을 목표로 실험 1에서는 인진쑥 및 녹차의 추출수율, 총 페놀화합물 함량, 항산화 활성, 항균 활성 및 면역 활성을 in vitro 실험을 통해 구명하는 한편, 실험 2에서는 이들 추출물을 육계에 음수 급여하여 생산성, 조직 및 소장의 변화, 장내 미생물 균총, 혈액 특성 및 비장 cytokines mRNA 발현량에 미치는 영향을 조사하였다. 인진쑥 및 녹차 추출물의 추출수율은 각각 16.8%, 26.5% 이었으며, 총 페놀화합물 함량은 15.28 및 26.74 GE mg/mL이었다. 또한 이들 추출물의 항산화, 항균 활성 및 닭의 splenocyte 증식에 미치는 영향을 조사한 결과, DPPH radical을 50% 감소시키는 농도인 $SC_{50}$은 각각 0.30 mg, 0.06 mg으로 나타냈으며, 유해균 3종(E. coli, S. aureus, S. Typhimurium)에 대해 항균 활성은 인진쑥 추출물이 33.7%, 21.6%, 28.5%, 녹차 추출물이 52.3%, 42.1%, 45.6%의 성장 저해율을 나타냈다. 닭의 splenocyte 증식에도 효과가 나타냈는데, 특히 인진쑥 추출물은 2.5 ${\mu}L$, 5.0 ${\mu}L$ 농도에서 림프구분열촉진인자로 알려진 Echinacea 및 concanavalin A보다 우수한 세포 증식도를 보였다. 실험 1에서 항산화, 항균 및 면역조절효과가 확인된 인진쑥, 녹차 추출물을 육계에 음수 급여한 결과, 인진쑥, 녹차 추출물 처리구가 무항생제 대조구(NC)에 비해 종료체중 및 증체량이 유의하게 증가하였으며(p<0.05), 맹장 내 Salmonella 수에서는 무항생제 대조구(NC)에 비해 유의하게 감소하거나, 감소하는 경향을 나타내었다(p<0.05). 혈액 생화학 조성에 있어서는 인진쑥, 녹차 추출물 처리시 총콜레스테롤, 간 및 신장의 이상 측정 지표인 AST 및 ALT가 대조구에 비해 유의하게 감소하였다(p<0.05). 본 연구를 통해 인진쑥 및 녹차 추출물의 생리활성효과를 구명하였으며, 이들 추출물은 육계 생산성을 향상시키고 유해균의 성장 및 증식을 억제시켜 장내 미생물 균총 안정화에 긍정적인 영향을 미치는 한편, 조직 발달 및 혈액 성상에 부정적인 영향을 미치지 않아 항생제 대체제로서의 이용 가능성을 확인하였다.

다국어 초록 (Multilingual Abstract)

Two experiments were conducted to determine whether water extracts from Artemisia capillaries (A. capillaries) and Camellia sinensis (C. sinensis) could be used as alternatives to antibiotic growth promoters in broiler feed. The experiment 1 was verified their chemical composition, extracts yields, total phenolic compounds concentration, antioxidant activity, antimicrobial activity, and chicken splenocytes proliferation through in vitro test. The extract yields of A. capillaries and C. sinensis were 26.5 and 16.8%, respectively. Total phenolic compounds concentrations of them expressed as gallic acid equivalent were 15.28 and 26.74 mg/mL, respectively. Electron donating abilities of them expressed as $SC_{50}$ showing 50% DPPH radical scavenging were 0.30 and 0.06 mg, respectively. Bacterial inhibitory rates of them against Escherichia coli, Staphylococcus aureus, and Salmonella Typhimurium were ranged from 42.1 to 52.3% and from 21.6 to 33.7%, respectively. And, these extracts increased proliferation of chicken splenocytes. Especially, A. capillaris was more excellent than Echinacea and Concanavalin A known as T-cell stimulator. The experiment 2 was investigated their effects on growth performance, relative organ weight, cecal microflora, blood biochemical parameters, and splenic cytokines mRNA expression in broiler chicks. Four hundred eighty 1-day-old male broiler chicks (Ross 308) were divided in to 4 treatment groups with 4 replicates of 30 birds in each group: NC (control, no antibiotics), PC (avilamycin, 10 ppm; salinomycin, 60 ppm), AC (A. capillaries, 100 ppm), and CS (C. sinensis, 100 ppm); treatments were administered through water supplementation. Final body weight was significantly higher in all treated groups than in NC (p<0.05). Cecal Salmonella numbers were significantly or somewhat decreased in all treated groups than in NC (p<0.05). The relative weights and lengths of the small intestine were more significantly decreased in the PC and AC groups than in the other groups. Cecal Salmonella numbers were significantly or somewhat decreased in all treated groups than in the NC group (p<0.05). The contents of total cholesterol, aspatate aminotransferase, and alanine aminotransferase in blood serum were more significantly decreased in all treated groups than in NC (p<0.05). In conclusion, these results suggested the possibility that these extracts could serve as alternatives for antibiotic growth promoters.

참고문헌 (Reference)

1 임정대, "한국산 약용식물로부터의 SOD 활성 및 Phenolic Compounds 함량 비교" 한국약용작물학회 12 (12): 191-202, 2004

2 한영희, "장내 미생물 개선효과가 있는 약용식물소재 탐색" 한국약용작물학회 15 (15): 26-29, 2007

3 김홍태, "인진쑥 추출물의 병원성 세균에 대한 시험관내 항균효과" 한국임상수의학회 24 (24): 130-136, 2007

4 장정연, "인진쑥 올리고당의 섭취가 비만쥐의 혈중 지질, 복강내 지방조직과 렙틴 수치에 미치는 영향" 한국영양학회 36 (36): 437-445, 2003

5 한은경, "에탄올 투여 흰쥐의 지질대사 및 간 독성 저하에 미치는 인진쑥 및 눈꽃동충하초의 영향" 한국식품영양과학회 38 (38): 1016-1023, 2009

6 정성제, "약용 식물 추출물의 항산화 활성 검색" 한국응용생명화학회 47 (47): 135-140, 2004

7 박창일, "쑥 분말의 급여가 계육의 저장기간 중 VBN, TBARS 및 지방산 조성에 미치는 영향" 한국축산식품학회 28 (28): 505-511, 2008

8 박성혜, "기능성 식품으로의 활용을 위한 한약자원에 관한 연구― (Ⅰ) 한국산 인진쑥의 영양성분 및 유효성분 중 Scopoletin 분석 ―" 동아시아식생활학회 13 (13): 552-560, 2003

9 최원균, "국내산 식물의 항균활성 검색" 한국식품영양학회 15 (15): 300-306, 2002

10 권혁신, "국내 자생 황금 추출물의 첨가 급여가 육계의 생산성, 면역 기능 및 장내 균총 변화에 미치는 영향" 한국가금학회 35 (35): 351-359, 2008

11 김용란, "국내 자생 약용식물자원(박하, 복분자 및 매리골드)의 첨가 급여가 육계의 생산성과 육질에 미치는 영향" 한국축산식품학회 29 (29): 168-177, 2009

12 김지혁, "겨우살이의 급여가 육계의 생산성 및 혈액 특성에 미치는 영향" 한국가금학회 34 (34): 129-136, 2007

13 Ahn, B. M., "What is In-Jin-Sook Artemisia capillaris, Artemisia iwayomogi, and Artemisia annua" 6 : 548-551, 2000

14 Moore, P. R., "Use of sufasuxidine, stroptothricine, and streptomycin in nutritional studies with the chick" 165 : 437-441, 1946

15 Windisch, W., "Use of phytogenic products as feed additives for swine and poultry" 86 : 140-148, 2008

16 Jamroz, D., "Use of active substances of plant origin in chicken diets based on maize and locally grown cereals" 46 : 485-493, 2005

17 Shan, B., "The in vitro antibacterial activity of dietary spice and medicinal herb extracts" 177 : 112-119, 2007

18 Trevisanato, S. I., "Tea and health" 58 : 1-10, 2000

19 Akpapunam, M. A., "Some physicochemical properties and anti-nutritional factors of raw, cooked and germinated Jack bean (Canvalia ensiformis)" 59 : 121-125, 1997

20 Chomczynski, P., "Single-step method of RNA isolation by acid guanidinium thiocyanate-phenolchloroform extraction" 162 : 156-159, 1987

21 Ahn, D. J., "Screening of herbal plant extracts showing antimicrobial activity against some food spoilage and pathogenic microorganisms" 8 : 109-116, 2000

22 SAS, "SAS/STAT Software for PC. Release 9.1, SAS Institute Inc., Cary, NC, USA"

23 Xue, M., "Review on research progress and prosperous of immune activities of bio-active polysaccharide" 3 : 15-18, 1996

24 Miyamoto, T., "Production and characterization of monoclonal antibodies detecting chicken interleukin-2 and the development of an antigen capture enzyme linked immunosorbent assay" 80 : 245-257, 2001

25 Greathead, H., "Plants and plant extracts for improving animal productivity" 279-290, 2003

26 Cowan, M. M., "Plant products as antimicrobial agents" 12 : 564-582, 1999

27 AOAC, "Official Methods of Analysis. 18th ed" Association of Official Analytical Chemist 931-, 2007

28 Seena, S., "Nutritional and anti-nutritional evaluation of raw and processed seeds of a wile legume, Canavalia cathartica of coastal sand dunes of India" 92 : 465-472, 2005

29 NRC, "Nutrient requirement of poultry. 9th ed" National Research Council, National Academy of Science 1994

30 Duncan, D. B., "Multiple range and multiple F test" 11 : 1-42, 1955

31 Ikeda, I., "Multifunctional effects of green tea catechins on prevention of the metabolic syndrome" 17 : 273-274, 2008

32 Lima, J. E., "Lymphocyte activation and cytokine production by Pisum sativum agglutinin (PSA) in vivo and in vitro" 41 : 147-155, 1999

33 Diaz, G. J., "Intoxication of Crotalaria pallid seeds to growing broiler chicks" 45 : 187-189, 2003

34 Farner, N. L., "Interleukin- 2: Molecular and clinical aspects. In : Cytokines in health and diseases" Marcel Dekker, Inc. 29-40, 1997

35 Hernandez, F., "Influence of two plant extracts on broilers performance, digestibility, and organ size" 83 : 169-174, 2004

36 Ma, D., "Influence of an aqueous extract of Ligustrum lucidum and an ethanol extract of Schisandara chinensis on parameters of antioxidative metabolism and spleen lymphocyte proliferation of broilers" 63 : 66-74, 2009

37 PARK, KAP JOO, "In Vitro Antiviral Activity of Aqueous Extracts from Korean Medicinal Plants Against Influenza Virus Type A" 한국미생물·생명공학회 15 (15): 924-929, 2005

38 Tonevitsky, A. G., "Immunotoxins containing A-chain of mistleotoe lectin É are more active than immunotoxins with ricin A-chain" 392 : 166-168, 1996

39 Feyes, A. L., "Green tea constituent epigallocatechin-3-gallate and induction of apotosis and cell cycle arrest in human carcinoma cell" 89 : 1881-1886, 1997

40 Son, S., "Free radical scavenging and antioxidant activity of caffeic acid amide and ester analogues: structure-activity relationship" 50 : 468-472, 2002

41 RDA, "Food composition Table. 7th ed" Rural Development of Administration 2006

42 Yang, C. S., "Effects of tea consumption on nutrition and health" 130 : 2127-2130, 2002

43 Lee, C. H., "Effects of mugwort water extracts on cadmimum toxicity in rats" 19 : 188-197, 1999

44 Stutz, M. W., "Effects of diet and bacitracin on body weight restriction on the intestine of broiler chicks" 62 : 1626-1632, 1983

45 Debersac, P., "Effects of a water soluble extract of rosemary and its purified component rosmarinic acid on xenobiotic-metabolizing enzymes in rat liver" 39 : 109-117, 2001

46 Guo, F. C., "Effects of a chinese herb medicine formulation, as an alternative for antibiotics on performance of broiler" 45 : 793-797, 2004

47 Dong, X. F., "Effect of polysavone (alfafa extract) on abdominal fat deposition and immunity in broiler chickens" 86 : 1955-1959, 2007

48 Muramatsu, K., "Effect of green tea catechins on plasma cholesterol level in cholesterol fed rats" 32 : 613-622, 1986

49 Garcia, V., "Effect of formic acid and plant extracts on growth, nutrient digestibility, intestine mucosa morphology, and meat yield of broilers" 16 : 555-562, 2007

50 Henry, P. R., "Effect of antibiotics on tissue trace mineral concentration and intestinal tract weight of broiler chicks" 66 : 1014-1018, 1987

51 C. J. Yang, "Effect of Green Tea By-product on Performance and Body Composition in Broiler Chicks" 아세아·태평양축산학회 16 (16): 867-872, 2003

52 Nam, S. M., "Effect of Artemisia iwayomogi Kitamura ethanol extract on lowering serum and liver lipid in rats" 27 : 338-343, 1998

53 Biswas, M. A. H., "Comparison of twodietary factors, green tea powder feeding and feed restriction, influencing laying performance and egg quality in hens" Mie Univ. 25 : 55-61, 2001

54 Singleton, V. L., "Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents" 16 : 144-158, 1965

55 Oliveira, J.T.A., "Changes in organs and tissues induced by feeding of purified kidney bean (Phaseolus vulgaris)" 8 : 943-947, 1988

56 Wang, R. J., "Can 2000 years of herbal medicine history help us solve problems in the year 2000? Biotechnology in the Feed Industry" 273-291, 1998

57 Gill, C., "Botanical feed additives" 21 : 14-17, 2000

58 Lee, S. D., "Bioactive constituents and utilities of Artemisa sp. as medicinal herb and foodstuff" 13 : 490-505, 2000

59 Lumeiji, J. T., "Avian clinical biochemistry. In: Clinical biochemistry of domestic animals" Academic Press 857-883, 1997

60 Blois, M. S., "Antioxidant determinations by use of a stable free radical" 181 : 1199-1200, 1958

61 Shan, B., "Antioxidant capacity of 26 spice extracts and characterization of their phenolic constituents" 53 : 7749-7759, 2005

62 Cai, Y. Z., "Antioxidant activity and phenolic compounds of 112 traditional Chinese medicinal plants associated with anticancer" 74 : 2157-2184, 2004

63 Shin, K. H., "Antimicrobial activities of volatile essential oils from Korean aromatic plants" 3 : 141-147, 1997

64 Kiso, Y., "Antihepatotoxic principles of Artemisia capillaris buds" 1 : 81-85, 1984

65 Hinton, M. H., "Antibiotics, poultry production and public health" 44 : 67-69, 1988

66 Hara-Kudo, Y., "Antibacterial activity of plants used cooking for aroma and taste" 67 : 2820-2824, 2004

67 Amarowicz, R., "Antibacterial activity of green tea polyphenols against Escherichia coli K12" 44 : 60-62, 2000

68 Lopponow, L. H., "Adult human vascular endothelial cells express the IL-6 gene differentially in response to LPS or IL-1" 122 : 493-503, 1989

69 Foster, W. H., "A practical evaluation of five food additives likely to be used as growth promoters in broiler rations" 13 : 123-131, 1972