Effects of garlic Allium sativum on the immune responses of olive flounder Paralichthys olivaceus

마늘, Allium sativum이 넙치, Paralichthys olivaceus의 면역반응에 미치는 영향

  • Lee, Jun-Hee (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Woo, Sung-Ho (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Eom, Yong-Hwan (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Hwang, Bun-Ok (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Kwon, Mun-Gyeong (Pathology Division, National Fisheries Research and Development Institute) ;
  • Bang, Jong-Deuk (Southeast Sea Fisheries Research Institute, National Fisheries Research and Development Institute) ;
  • Park, Soo-Il (Aquatic Life Medicine, Pukyong National University)
  • 이준희 (부경대학교 수산생명의학과) ;
  • 우승호 (부경대학교 수산생명의학과) ;
  • 엄용환 (부경대학교 수산생명의학과) ;
  • 황분옥 (부경대학교 수산생명의학과) ;
  • 권문경 (국립수산과학원 병리연구과) ;
  • 방종득 (국립수산과학원 남동해수산연구소) ;
  • 박수일 (부경대학교 수산생명의학과)
  • Received : 2010.02.23
  • Accepted : 2010.04.12
  • Published : 2010.04.30

Abstract

This study was aimed to investigate the effects of injection of garlic, Allium sativum, extract and immersion in garlic juice on the nonspecific immunity and the resistance against the artificial infection of Streptococcus iniae and Edwardsiella tarda of the olive flounder, Paralichthys olivaceus. The nonspecific immune mechanisms were assessed in terms of lysozyme activity, nitroblue-tetrazolium (NBT) assay and superoxide dismutase (SOD) activity etc. Relative percent survival (RPS) was assessed by the challenge with S. iniae BS10 or E. tarda KE-1. Almost of the garlic extract injected groups showed the enhanced level of the tested nonspecific immune factors. In the challenge test with S. iniae and E. tarda, RPS of 5% garlic extract pre-injected group was much higher than that of any other tested groups, respectively. Almost of the garlic juice immersion tested groups exhibited strengthened nonspecific immune defence factors, lysozyme activity, the number of lymphocytes and neutrophils, NBT reduction and SOD activity in kidney. In the challenge with S. iniae and E. tarda, RPS in the 0.25 g/L of garlic juice immersed group was much higher than any other tested groups, respectively. The results suggest that the garlic extract and juice would be effective to enhance the nonspecific immunity and protective ability of olive flounder against fish disease such as S. iniae and E. tarda.

본 연구에서는 마늘 추출물과 착즙액이 넙치의 비특이적 면역력에 미치는 영향과 투여 방법에 따른 효과를 밝히고자 하였다. 마늘 추출물의 투여는 복강 주사법을, 마늘 착즙액은 침지법을 사용하였다. 마늘 착즙액 침지법은 일정 농도의 착즙액을 해수에 현탁하여 2일 간격으로 3회 침지하였다. 병원균의 공격 시험은 시험어에 마늘 추출물의 일정량을 2일 간격으로 3회 주사한 지2일 후에 실시하였다. 그 결과, 마늘 추출물의 복강 주사 시험구 중 5% 투여구에서 lysozyme의 활성이 가장 유의적인 증가를 나타내었다. 호중구의 수는 1, 5 및 10%를 주사하였을 때 유의적인 증가를 보였다 (p<0.05). SOD activity는 신장에서는 1, 3 및 5% 시험구에서, 간에서는 1 및 5% 시험구가 대조구에 비해 유의적인 증가를 나타내었다 (p<0.05). NBT 환원 시험은 3 및 5% 시험구에서 높은 NBT 환원량을 나타내었다. 마늘 착즙액의 침지 시험에서는 0.25 g/L 시험구에서 체표 점액 lysozyme 활성과 혈청 lysozyme 활성이 가장 높은 유의적인 증가를 하였다. 림프구와 호중구의 수는 0.25, 0.5 및 1.0 g/L 시험구에서 대조구에 비해 유의적인 증가를 보였다 (p<0.05). SOD activity는 신장에서는 모든 시험구에서, 간에서는 0.1, 0.25 및 0.5 g/L 시험구에서 대조구에 비해 유의적인 증가를 나타내었다 (p<0.05). NBT 환원 시험은 0.1, 0.25 및 0.5% 시험구에서 대조구와 비교하여 유의적인 증가를 나타내었다 (p<0.05). 시험구의 생존율은 S. iniae BS10과 E. tarda KE-1의 공격에 대하여 5% 마늘 추출물 복강 투여와 0.25 g/L 마늘 착즙액 침지 시험구의 상대생존율이 가장 높았다. 이상의 결과를 종합하여 볼 때 마늘 추출물과 착즙액이 넙치에 면역능 향상과 어류 질병에 저항성 증강에 효과가 있는 것으로 사료된다.

Keywords

References

  1. Alnaqeeb, M.A., Thomson, M., Bordia, T. and Ali, M.: Histopathological effects of garlic onliver and lung of rats. Toxicol. Lett. 85:157-164, 1996. https://doi.org/10.1016/0378-4274(96)03658-2
  2. Aoki, T.: Present and future problems concerning the development of drug resistance in Aquaculture, In Chemotherapy in Aquaculture :From Theory to Reality, Office International Des Epizooties. pp.254-262, Paris, France, 1992.
  3. Cavallito, C.J., Buck, S.J. and Suter, C.M.: Allicin, the antibacterial principle of Allium sativum. I. Isolation, physical properties and antibacterial action. J. Am. Chem. Soc., 66:1950-1951, 1944. https://doi.org/10.1021/ja01239a048
  4. Cavallito, C.J. and Bailey, J.H.: Allicin, the antibacterial principle of Allium sativum. II. Determination of the chemical structure. J. Am. Chem. Soc., 66:1952-1954, 1944. https://doi.org/10.1021/ja01239a049
  5. Chang, C.F., Su, M.S., Chen, H.Y. and Liao, I.C.: Dietary $\beta$-1,3-glucan effectively improves immunity and survival of Penaeus monodon challenged with white spot syndrome virus. Fish Shellfish Immunol., 15:297-310, 2003 https://doi.org/10.1016/S1050-4648(02)00167-5
  6. Chen, D. and Ainsworth, A.J.: Glucan administration potentiates immune defense mechanism of channel catfish, Ictalurus punatatus Rafinesque. J. Fish Dis., 15:295-304, 1992. https://doi.org/10.1111/j.1365-2761.1992.tb00667.x
  7. Choi, M.S., Park, S.W. and Park, K.H.: Effect of red ginseng extract on immune function of israeli carp, Cyprinus carpio. J. Fish Pathol., 18:277-285, 2005.
  8. Chrisybapita, D., Divyagnaneswari, M. and Dinakaran, M.R.: Oral administration of Eclipta alba leaf aqueous extract enhances the non-specific immune responses and disease resistance of Oreochromis mossambicus. Fish Shellfish Immunol., 23:840-852, 2007. https://doi.org/10.1016/j.fsi.2007.03.010
  9. Couso, N., Castro, R., Noya, M., Obach, A. and Lamas, J.: Location of superoxide production sites in turbot neutrophils and gilthead seabream acidophilic granulocytes during phagocytosis of glucan particles. Develop. Comp. Immunol., 25:607-618, 2001. https://doi.org/10.1016/S0145-305X(01)00019-2
  10. Delaha, E.C. and Garagusi, V.F.: Inhibition of mycobacteria by garlic extract (Allium sativum). Antimicrob. Agents Chemother., 27:485-486, 1985. https://doi.org/10.1128/AAC.27.4.485
  11. Dina, A.K., Bhathena, Z.P., Sahu, N.P, Jha, A. and Mukherjee, S.C.: Dietary microbial levan enhances cellular non-specific immunity and survival of common carp (Cyprinus carpio) juveniles. Fish Shellfish Immunol., 22:477-486, 2007. https://doi.org/10.1016/j.fsi.2006.06.005
  12. Divyagnaneswari, M., Christybapita, D. and Dinakaran, M.R.: Enhancement of nonspecific immunity and disease resistance in Oreochromis mossambicus by Solanum trilobatum leaf fractions. Fish Shellfish Immunol, 23:249-259, 2007. https://doi.org/10.1016/j.fsi.2006.09.015
  13. Ellis, A.E.: Immunity to bacteria in fish. Fish Shellfish Immunol., 9:291-308, 1999. https://doi.org/10.1006/fsim.1998.0192
  14. Engstad, R.E., Robertson, B. and Frivold, E.: Yeast glucan induce increase in activity of lysozyme and complement-mediated haemolytic activity in Atlantic salmon blood. Fish Shellfish Immunol., 2:287-297, 1992. https://doi.org/10.1016/S1050-4648(06)80033-1
  15. Eo, J and L. K.J.: Effect of dietary ascorbic acid on growth and non-specific immune responses of tiger puffer, Takifugu rubripes. Fish Shellfish Immunol., 25:611-616, 2008. https://doi.org/10.1016/j.fsi.2008.08.009
  16. Ghannoum, M.A.: Studies on the anticandidalmode of action of Allium sativum (garlic). J. Gen. Microbiol., 134:2917-2924, 1988.
  17. Grondel, J.L., Nouws, J.F.M., DeJong M., Schutte, A.R., and Driessens, F.: Pharmacokinetics and tissue distribution of oxytetracycline in carp, Cyprinus carpio L., following different routes of administration. J. Fish Dis., 10:153-163, 1987. https://doi.org/10.1111/j.1365-2761.1987.tb01057.x
  18. Hong, Y.S., Ham, Y.A., Choi, J.H., and Kim, J.: Effects of allylsulfur compounds and garlic extract on the expression of Bcl-2, Bax, and p53 in non small cell lung cancer cell lines. Exp. Mol. Med., 32:127-134, 2000. https://doi.org/10.1038/emm.2000.22
  19. Jorgensen, J.B., Lunde, H. and Robertsen, B.: Peritoneal and head kidney cell response to intraperitoneally injected yeast glucan in Atlantic salmon, Salmo salar L. J. Fish Dis., 16:313-325, 1993a. https://doi.org/10.1111/j.1365-2761.1993.tb00865.x
  20. Jorgensen, J.B., Sharp, G.J.E., Secombes, C.J. and Robertsen, B.: Effect of a yeast-cell wall glucan on the bactericidal activity of rainbow trout macrophages. Fish Shellfish Immunol., 3:267-277, 1993b. https://doi.org/10.1006/fsim.1993.1026
  21. Jung, S.H., Lee, J.S., Han, H.K., Jun, C.Y. and Lee, H.Y.: Effects of medical herb extract on non specific immune response, haematology and disease resistance on Olive flounder, Paralichthys olivaceus by oral administration. J. F. Pathol., 15:25-35, 2002.
  22. Kajita, Y., Sakai, M., Kobayashi, M. and Kawaushi, H.: Enhancement of non-specific cytotoxic activity of leucocytes in rainbow trout Oncorhynchus mykiss injected with growth hormone. Fish Shellfish Immunol., 2:155-157, 1992. https://doi.org/10.1016/S1050-4648(05)80044-0
  23. Kawakami, H., Hiratsuka, M. and Dosako, S.: Effects of iron- saturated lactoferrin on iron absorption. Agric. Biol. Chem., 52:903-908, 1998.
  24. Kitao, T., Yoshida, T., Anderson, D.P., Dixon, O.W. and Blanch, A.: Immunostimulation of antibody-producing cells and humoral antibody to fish bacterins by a biological response modifier. J. Fish Biol., 31:87-91, 1987. https://doi.org/10.1111/j.1095-8649.1987.tb05298.x
  25. Kim, K.H., Hwang, Y.J. and Bai, S.C.: Enhancement of chemiluminescent response of phagocytic cells from juvenile Rockfish, Sebastes schlegeli, by oral administration of levamisole. J. Fish Sci. Tech., 1:42-47, 1998.
  26. Kodama, H., Mukamoto, N., Baba, T. and Mule, D.M.: Macrophage-colony stimulating activity in rainbow trout Oncorhynchus mykiss serum. In: J.S. Stolen and T.C. Fletcher (eds) Modulators of fish immune responses, Vol. 1. pp.59-66, SOS Publication, Fair Haven, NJ, 1994.
  27. Miles, A.A. and Misra S.S.: The estimate of the bacterial power of the blood. J. Hygiene, 38:873-885, 1938.
  28. Misra, C.K., Das, B.K., Mukherjee, S.C. and Meher, P.K.: The immunomodulatory effects of tuftsin on the non-specific immune system of indian major carp, Labeo rohita. Fish Shellfish Immunol., 20:728-738, 2006a. https://doi.org/10.1016/j.fsi.2005.09.004
  29. Misra, C.K., Das, B.K., Mukherjee, S.C. and Pattnaik. P.: Effect of long term administration of dietary $\beta$-glucan on immunity, growth and survival of Labeo rohita fingerlings. Aquaculture, 255:82-94, 2006b. https://doi.org/10.1016/j.aquaculture.2005.12.009
  30. Murray, C.K. and Fletcher, T.C.: The immunohistochemical location of lysozyme in plaice (Pleuronectes platessa L.) tissue. J. Fish. Biol., 9:329-334, 1976. https://doi.org/10.1111/j.1095-8649.1976.tb04681.x
  31. Nakagawa, H., Tsuta, K., Kiuchi, K., Senzaki, H., Tanaka, K. and Hioki, A.: Growth inhibitory effects of diallyl disulfide on human breast cancer cell lines. Carcinogenesis, 22:891-897, 2001. https://doi.org/10.1093/carcin/22.6.891
  32. Ordas, M.C., Novoa, B. and Figueras, A.: Modulation of the chemiluminescence response of mediterranean mussel (Mytilus galloprovincialis) haemocytes. Fish Shellfish Immunol., 10:611-622, 2000. https://doi.org/10.1006/fsim.2000.0276
  33. Parry, R.M., Chandau, R.C. and Shahani, R.M.: A rapid and sensitive assay of muramidase. Proc. Soc. Exp. Biol. Med., 119:384-386, 1965. https://doi.org/10.3181/00379727-119-30188
  34. Raa, J., Torstad, G., Engstad, R. and Robertsen, B.:The use of immunostimulants to increase resistance of aquatic organisms to microbial infections. In: M. Sharif, R. P. Subasighe and Arthur, J. R. (eds.), Dis. Asian A. Vol. 1. pp.39-50, Fish Health Section, Asian Fish. Soc. Manila, Philippines, 1993.
  35. Rairakhwada, D., Pal, A.K., Bhathena, Z.P., Sahu, N.P., Jha, A. and Mukherjee, S.C.: Dietary microbial levan enhances cellular non-specific immunity and survival of common carp (Cyprinus carpio) juveniles. Fish Shellfish Immunol., 21:346-356, 2007.
  36. Sakai, M.: Current research status of fish immunostimulants. Aquaculture, 172:63-92, 1999. https://doi.org/10.1016/S0044-8486(98)00436-0
  37. Sheen, L.Y., Li, C.K., Sheu, S.F., Meng, R.H.C. and Tsai, S.J.: Effect of the active principle of garlic-diallyl sulfide- on cell viability, detoxification capability and the antioxidation system of primary rat hepatocytes. Food Chem. Toxicol., 34:971-978.
  38. Siwicki, A.K., Anderson, D.P. and Dixon, O.W.: In vitro immunostimulation of rainbow trout Oncohynchus mykiss spleen cells with lavamisole. Dev. Comp. Immunol., 14:231-237, 1990. https://doi.org/10.1016/0145-305X(90)90094-U
  39. Soffar, S.A. and Mokhtar, G.M.: Evaluation of the antiparasitic effect of aqueous garlic (Allium sativum) extract in Hymenolepiasis nana and giardiasis. J. Egypt. Soc. Parasitol., 21:497-502, 1991.
  40. Solem, S.T., Jorgensen, J.B. and Robertsen, B.:Stimulation of respiratory burst and phagocytic activity in Atlantic salmon Salmo salar L. macrophages by lipopolysaccharide. Fish Shellfish Immunol., 5:475-491, 1995. https://doi.org/10.1016/S1050-4648(95)80049-2
  41. Takahashi, Y, Itami, T. and Konegawa K.: Enzymatic properties of partially lysozyme from the skin mucus of carp. Bull. Jap. Soc. Sci. Fish., 52:1209-1214, 1986. https://doi.org/10.2331/suisan.52.1209
  42. Uchida, Y., Takehaski, T. and Sato, N.: The characteristics of the antibacterial activity of garlic. Jpn. J. Antibiot., 28:638-642, 1975.
  43. Villamil, L., Figueras, A. and Novoa, B.: Immunomodulatory effects of nisin in turbot (Scophthalmus maximus L.). Fish Shellfish Immunol., 14:157-69, 2003. https://doi.org/10.1006/fsim.2002.0425
  44. Wu, C.C., Chung, J.G., Tsai, S.J., Yang, J.H. and Sheen, L.Y.: Differential effects of allyl sulfides from garlic essential oil on cell cycle regulation in human liver tumor cells. Food Chem. Toxicol., 42:1937-1947, 2004. https://doi.org/10.1016/j.fct.2004.07.008
  45. Yano, T., Mangindaan, R.E.P. and Matsuyama, H.:Enhancement of the resistance of carp Cyprinus carpio to experimental Edwardsiella tarda infection, by some $\beta$-1,3 glucans. Nippon Suisan Gakkaishi, 55:1815-1819, 1991.
  46. Yeh, S.P., Chang, C.A., Chang, C.Y., Liu, C.H. and Cheng, W.: Dietary sodium alginate administration affects fingerling growth and resistance to Streptococcus sp. and iridovirus, and juvenile non-specific immune responses of the orange-spotted grouper, Epinephelus coioides. Fish Shellfish Immunol., 25:19-27, 2008. https://doi.org/10.1016/j.fsi.2007.11.011
  47. 김진우, 박수일, 전세규: 양식 넙치로부터의 lysozyme 정제와 어류 병원성 세균에 대한 정균 작용. 한국어병학회지, 5:87-92, 1992
  48. 우승호, 김현정, 이주석, 김진우, 박수일: 해수양식 어류에서 분리된 연쇄상구균의 종류와 병원성. 한국어병학회지, 19:17-33, 2006.
  49. 이덕찬, 김이청, 김진우, 박수일: Edwardsiella tarda extracellular products (ECPs)의 인위 투여에 따른 넙치, Paralichthys olivaceus의 생체 반응. 한국어병학회지, 18:215-225, 2005.
  50. 유병화, 박수일, 전세규: 어류혈청의 보체에 의한 살균작용. 한국어병학회지, 5:9-18, 1992.