DOI QR코드

DOI QR Code

Antimicrobial Effect of Chitosan and Chitooligosaccharides against Bacterial Diseases of Cultured Flounder

양식 넙치의 세균성 질병에 대한 키토산 및 키토올리고당의 항균효과

  • 양병규 (제주대학교 해양생산과학부) ;
  • 이제희 (제주대학교 해양생산과학부) ;
  • 김수현 (제주대학교 식품공학과) ;
  • 전유진 (제주대학교 해양생산과학부)
  • Published : 2004.02.01

Abstract

Antimicrobial effect of chitosan and its oligosaccharides was examined on Vibrios sp., Edwardsiella tarda and Streptococcus sp., which are major pathogenic bacteria inducing bacterial diseases of acquacultured flounder. Chitosan oligosaccharides (COS ) were produced by enzymatic hydrolysis of chitosan in an ultrafiltration mombrane bioreactor system which was established with three membranes with different molecular weight cut-off (MWCO) 1,000, 5,000 and 10,000, and fractionated into three kinds of COS, based on their molecular weight sizes. The three kinds of COS were as follows : relatively high molecular weight COS [HMW-COS, molecular weight distribution of 7,000 to 24,000 Da〕, medium molecular weight COS 〔MMW-COS, 1,500 to 6,000 Da〕, and low molecular weight COS 〔LMW-COS, 1,000 to 1,500 Da). Chitosan and HMW-COS effectively inhibited the growths of Vibrio sp. and Streptococcus sp. and their antimicrobial activities were superior to the others with smaller molecular weights. This result suggested that antimicrobial effect of chitosan preparations extremely depend on their molecular weight sizes. Antimicrobial effect of chitosan and HMW-COS on E. tarda was improved by longer inoculation times. Scanning electron microscopy in morphological change of E. tarda treated with chitosan preparations showed that chitosan and HMW-COS bound to the cells and suppressed the growth of the cells. This observation appears to prove the fact that positive charged amines of chitosan electrostatically bind to negative charged compounds of cell walls.

양식넙치의 세균성 질병의 원인균주인 Vibrio sp., Edwardsiella tarda 및 Streptococcus sp.를 제주산 양식 넙치로부터 직접 분리하였으며, 이들 세균에 대하여 키토산 및 분자량 크기에 따라 분획된 3종류의 키토산올리고당(HMW-COS, MMW-COS, LMW-COS)의 항균활성을 검토하였다. 키토산과 HMW-COS는 모든 세균에 대한 항균활성이 우수하였으며, 특히 Vibrio sp.과 Sfreptococcus sp.에 대해서 높은 항균활성을 나타내었다. 이들에 대한 결과는 키토산의 항균활성은 분자량에 따라 크게 의존한다는 사실을 보여주었다 E.tarda에 대한 키토산 및 HMW-COS의 항균활성은 처리시간을 증가시킴으로써 항균활성이 증가함을 보였으며, 이들의 항균기 작을 조사하기 위 한 SEM의 관찰에서, 세균들은 키토산에 의해 응집되어 서로 엉켜있는 형태를 보여주고 있었다. 이러한 현상은 키토산 및 HMW-COS가 세균의 증식을 명확하게 억제하고 있음을 보여주는 증거라 할 수 있다. 이것은 지금까지 여러 보고에서 밝혀진 키토산의 항균활성 메카니즘과 일치하는 것이라 판단된다.

Keywords

References

  1. Jia X, Patrzykat A, Devlin RH, Ackerman PA, Iwama GK, Hancock REW. 2000. Antimicrobial peptides protect coho salmon from Vibrio anguillarum infections. Appl Environ Microbiol 66: 1928-1932. https://doi.org/10.1128/AEM.66.5.1928-1932.2000
  2. Arvanitoyamis IS, Nakayama A, Aiba S. 1998. Chitosan and gelatin based edible films: state diagrams, mechanical and permeation properties. Carbohydr Polym 37: 371-382. https://doi.org/10.1016/S0144-8617(98)00083-6
  3. Rosales-Cortes M, Peregrina-Sandoval J, Banuelos-Pineda J, Sarabia-Estrada R, Gomez-Rodiles CC, Albarran-Rodriguez E, Zaitseva GP, Pita-Lopez ML. 2003. Immunological study of a chitosan prosthesis in the sciatic nerve regeneration of the axotomized dog. J Biomater Appl 18: 15-23. https://doi.org/10.1177/0885328203018001002
  4. Risbud MV, Bhonde MR, Bhonde RR. 2001. Effect of chitosan-polyvinyl pyrrolidone hydrogel on proliferation and cytokine expression of endothelial cells: implications in islet immunoisolation. J Biomed Mater Res 57: 300-305 https://doi.org/10.1002/1097-4636(200111)57:2<300::AID-JBM1171>3.0.CO;2-Q
  5. van der Lubben IM, Verhoef JC, Borchard G, Junginger HE. 2001. Chitosan and its derivatives in mucosal drug and vaccine delivery. Eur J Pharm Sci 14: 201-207. https://doi.org/10.1016/S0928-0987(01)00172-5
  6. van der Lubben IM, Verhoef JC, Borchard G, Junginger HE. 2001. Chitosan for mucosal vaccination. Adv Drug Deliv Rev 52: 139-144. https://doi.org/10.1016/S0169-409X(01)00197-1
  7. Westerink MA, Smithson SL, Srivastava N, Blonder J, Coeshott C, Rosenthal GJ. 2001. ProJuvant (Pluronic F127/chitosan) enhances the immune response to intranasally administered tetanus toxoid. Vaccine 20: 711-723. https://doi.org/10.1016/S0264-410X(01)00423-6
  8. Kobayashi M, Watanabe T, Suzuki S, Suzuki M. 1990. Effect of N-acetylchitohexaose against Candida albicans infection of tumor-bearing mice. Microbiol Immunol 34: 413-426. https://doi.org/10.1111/j.1348-0421.1990.tb01024.x
  9. Tokoro A, Suzuki K, Matsumoto T, Mikami T, Suzuki S, Suzuki M. 1988. Chemostatic response of human neutrophils to N-acetylchitohexaose in vitro. Microbiol Immunol 32: 387-395. https://doi.org/10.1111/j.1348-0421.1988.tb01398.x
  10. Jeon YJ, Kim SK. 2002. Antitumor activity of chitosan oligosaccharides produced in ultrafiltration membrane reactor system. J Microbiol Biotechnol 12: 503-507.
  11. Qin C, Du Y, Xiao L, Li Z, Gao X. 2002. Enzymic preparation of water-soluble chitosan and their antitumor activity. Int J Biol Macromol 31: 111-117. https://doi.org/10.1016/S0141-8130(02)00064-8
  12. Kato Y, Onishi H, Machida Y. 2000. Biological fate of highlysuccinylated N-succinyl-chitosan and antitumor characteristics of its water-soluble conjugate with mitomycin C at i.v. and i.p. administration into tumor-bearing mice. Biol Pharm Bull 23: 1497-1503. https://doi.org/10.1248/bpb.23.1497
  13. Sato M, Onishi H, Takahara J, Machida Y, Nagai T. 1996. In vivo drug release and antitumor characteristics of watersoluble conjugates of mitomycin C with glycol-chitosan and N-succinyl-chitosan. Biol Pharm Bull 19: 1170-1177. https://doi.org/10.1248/bpb.19.1170
  14. Tsukada K, Matsumoto T, Aizawa K, Tokoro A, Naruse RS, Suzuki S, Suzuki M. 1990. Antimetastatic and growthinhibitory effects of N-acetylchitohexaose in mice bearing Lewis lung carcinoma. Jpn J Cancer Res 81: 259-265. https://doi.org/10.1111/j.1349-7006.1990.tb02559.x
  15. Tokoro A, Tatewaki N, Suzuki K, Mikami K, Suzuki S, Suzuki M. 1988. Growth-inhibitory effect of hexa-Nacetylchitohexaose and chitohexaose against Meth-A solid tumor. Chem Pharm Bull 36: 784-790. https://doi.org/10.1248/cpb.36.784
  16. Kim KW, Thomas RL, Lee C, Park HJ. 2003. Antimicrobial activity of native chitosan, degraded chitosan, and O-carboxymethylated chitosan. J Food Prot 66: 1495-1498.
  17. Rabea EI, Badawy ME, Stevens CV, Smagghe G, Steurbaut W. 2003. Chitosan as antimicrobial agent: applications and mode of action. Biomacromolecules 4: 1457-1465. https://doi.org/10.1021/bm034130m
  18. Savard T, Beaulieu C, Boucher I, Champagne CP. 2002. Antimicrobial action of hydrolyzed chitosan against spoilage yeasts and lactic acid bacteria of fermented vegetables. J Food Prot 65: 828-833.
  19. Jeon YJ, Kim SK. 2001. Effect of antimicrobial activity by chitosan oligosaccharides N-conjugated with asparagine. J Microbiol Biotechnol 11: 281-286.
  20. Jeon YJ, Park PJ, Kim SK. 2001. Antimicrobial effect of chitooligosaccharides produced by bioreactor. Carbohydr Polym 44: 71-76. https://doi.org/10.1016/S0144-8617(00)00200-9
  21. Jeon YJ, Kim SK. 2000. Production of chitooligosaccharides using an ultrafiltration membrane reactor and their antibacterial activity. Carbohydr Polym 41: 133-141. https://doi.org/10.1016/S0144-8617(99)00084-3
  22. Roller S, Covill N. 1999. The antifungal properties of chitosan in laboratory media and apple juice. Int J Food Microbiol 47: 67-77. https://doi.org/10.1016/S0168-1605(99)00006-9
  23. Hadwiger LA, Ogawa T, Kuyama H. 1994. Chitosan polymer sizes effective in inducing phytoalexin accumulation and fungal suppression are verified with synthesized oligomers. Mol Plant Microbe Interact 7: 531-533. https://doi.org/10.1094/MPMI-7-0531
  24. Young DH, Kohle H, Kauss H. 1982. Effect of chitosan on membrane permeability of suspension-cultured Glycine max and Phaseolus vulgaris cells. Plant Physiol 70: 1449-1454. https://doi.org/10.1104/pp.70.5.1449
  25. Hadwiger LA, Beckman JM, Adams MJ. 1981. Localization of fungal components in the pea-Fusarium interaction detected immunochemically with anti-chitosan and antifungal cell wall antisera. Plant Physiol 67: 170-175. https://doi.org/10.1104/pp.67.1.170
  26. Kendra DF, Hadwiger LA. 1984. Characterization of the smallest chitosan oligomer that is maximally antifungal to Fusarium solani and elicits pisatin formation in Pisum sativum. Exp Mycol 8: 276-281. https://doi.org/10.1016/0147-5975(84)90013-6
  27. Jeon YJ, Kim SK. 2001. Potential immuno-stimulating effect of antitumoral fraction of chitosan oligosaccharides. J Chitin Chitosan 6: 163-167.
  28. Jeon YJ, Kim SK. 2000. Continuous production of chitooligosaccharides using a dual reactor system. Process Biochem 35: 623-632. https://doi.org/10.1016/S0032-9592(99)00118-1
  29. Miyoshi S. 1989. Infection of Vibrio sp. J Antibacterial Antifungi 17: 279-285.
  30. Kim KH, Choi DL, Chung JK, Chun SK. 1992. Experimental infection of Edwardsiella tarda in the tilapia. J Fish Pathol 5: 61-76.
  31. Heo GJ, Lee YS. 1996. Efficacy of clindamycin for the control of streptococcal infection in cultured fish, flounder fish (Paralichthys olivaceus) and eel (Anguilla japonica). Kor J Lab Ani Sci 12: 25-30.
  32. Jung SH, Kim JW. 2000. In vitro antimicrobial activity in combination of antibacterials against fish-pathogenic bacteria. J Fish Pathol 13: 45-51.
  33. Hasan Jama Y, Varadaraj MC. 1999. Antibacterial effect of plantaricin LP84 on foodborne pathogenic bacteria occurring as contaminants during idli batter fermentation. World J Microbiol Biotechnol 15: 27-32. https://doi.org/10.1023/A:1008887201516
  34. Heo GJ, Lee JH. 1994. A study on efficacy and safety of gentamicin to bacterial diseases in cultured fish, Cryprinus carpio and Paralichthys olivaceus. Kor J Vet Publ Hlth 18: 327-331.
  35. Jung SH, Sohn YC, Kim YC. 2001. In vitro effect of water extract of medicinal herbs on antimicrobial activity against fish pathogenic bacteria and superoxide production of kidney phagocytes in olive flounder, Paralichthys olivaceus. J Fish Pathol 14: 3-10.
  36. Cho SH, Seo IW, Choi JD, Chun SS, Na TK, Chung SK, Kang DH. 1992. Disinfectant and inhibitory effect of natural antimicrobial agent on Vibrio vulnificus in fish. Kor J Food Hygiene 7: 99-106.
  37. Jung SH, Lee JS, An HK, Jun CY, Lee HY. 2002. Effect of medicinal herb extract on non-specific immune responses, hematology and disease resistance on olive flounder, Paralichthys olivaceus by oral administration. J Fish Pathol 15: 25-35.
  38. Kwon MG, Lee YH, Park SU, Kim BS, Park SI. 2002. The effect of charcoal in diet on the immune responses of flounder, Paralichthy olivaceus. J Fish Pathol 15: 17-24.
  39. Park SW, Kim YG, Choi DL. 1997. Enhancement of bacterial disease resistance in rockfish (Sebastes schlegeli) by ${\beta}$-glucan administration. J Fish Pathol 10: 143-152.
  40. Kim SR, Kim SH. 2002. The effect of food treated with gamma radiation after inoculation with pathogenic bacteria in the flounder (Paralichthys olivaceus). J Vet Clin 19: 7-13.
  41. Liltved H, Landfald B. 2002. Effect of high intensity light on ultraviolet-irradiated and non-irradiated fish pathogenic bacteria. Wat Res 34: 481-486. https://doi.org/10.1016/S0043-1354(99)00159-1
  42. Yang BG, Jeon YJ, Heo MS. 2003. Screening and characterization of probiotic strains for prevention of bacterial fish diseases. Kor J Microbiol Biotechnol 31: 129-134.
  43. Sugita H, Okano R, Suzuki Y, Iwai D, Mizukami M, Akiyama N, Matsuura S. 2002. Antibacterial abilities of intestinal bacteria from larval and juvenile Japanese flounder against fish pathogens. Fish Sci 68: 1004-1011. https://doi.org/10.1046/j.1444-2906.2002.00525.x
  44. No HK, Park NY, Lee SH, Meyers SP. 2002. Antibacterial activity of chitosans and chitosan oligomers with different molecular weights. Int J Food Microbiol 74: 65-72. https://doi.org/10.1016/S0168-1605(01)00717-6
  45. Kim SK, Jeon YJ, Zan HC. 2000. Antibacterial effect of chitooligosaccharides with different molecular weights prepared using membrane bioreactor. J Chitin Chitosan 5: 1-8.
  46. Tsai GJ, Wu ZY, Su WH. 2000. Antibacterial activity of a chitooligosaccharide mixture prepared by cellulase digestion of shrimp chitosan and its application to milk preservation. J Food Prot 63: 747-752.
  47. Choi BK, Kim KY, Yoo YJ, Oh SJ, Choi JH, Kim CY. 2001. In vitro antimicrobial activity of a chitooligsaccharide mixture against Actinobacillus actinomycetemcomitans and Streptococcus mutans. Int J Antimicrobiol Agents 18: 553-557. https://doi.org/10.1016/S0924-8579(01)00434-4
  48. Tsai GJ, Su WH, Chen HC, Pan CL. 2002. Antimicrobial activity of shrimp chitin and chitosan from different treatments and applications of fish preservation. Fish Sci 68: 170-177. https://doi.org/10.1046/j.1444-2906.2002.00404.x

Cited by

  1. Hepatic and skeletal muscle mitochondrial toxicity of chitosan oligosaccharides of normal and diabetic rats vol.26, pp.9, 2016, https://doi.org/10.1080/15376516.2016.1222643
  2. Review of Recent Studies and Research Analysis for Anti-oxidant and Anti-aging Materials vol.14, pp.4, 2016, https://doi.org/10.20402/ajbc.2016.0107
  3. 초산 세척과 키토산 첨가에 의한 간장게장의 미생물학적 품질 향상 vol.34, pp.3, 2019, https://doi.org/10.13103/jfhs.2019.34.3.296