Journal of fish pathology (한국어병학회지)

The Korean Society of Fish Pathology (한국어병학회)
권호 표지
DOI QR코드

DOI QR Code

Antibacterial Effects of Pseudomonas aeruginosa MB I-3 against Listonella anguillarum

어류 병원세균, Listonella anguillarum에 대한 Pseudomonas aeruginosa MB I-3의 항균 효과

  • Lee, Su-Jung (Busan Marine Resources Research Institute) ;
  • Youn, I Na (Department of Aqualife Medicine, Chonnam National University) ;
  • Kim, Jin-Do (Pathology division, National Fisheries Research and Development Institute) ;
  • Lee, Jung Sick (Department of Aqualife Medicine, Chonnam National University) ;
  • Kim, Eunheui (Department of Aqualife Medicine, Chonnam National University)
  • 이수정 (부산광역시 수산자원연구소) ;
  • 윤이나 (전남대학교 수산생명의학과) ;
  • 김진도 (국립수산과학원 병리연구과) ;
  • 이정식 (전남대학교 수산생명의학과) ;
  • 김은희 (전남대학교 수산생명의학과)
  • Received : 2013.12.20
  • Accepted : 2014.04.11
  • Published : 2014.04.30
Download PDF
⟨ Previous Next ⟩

Abstract

To study the possible use of probiotics in fish farming, The in vitro and in vivo antibacterial effects of Pseudomonas aeruginosa MB I-3 (MB I-3) against the fish pathogenic bacterium Listonella anguillarum were evaluated. The inhibitory effects of MB I-3 against vibrios were investigated by the double layer method and the co-culture. The results showed that MB I-3 inhibited the growth of pathogenic vibrios including Listonella anguillarum, Vibrio alginolyticus, Vibrio cholerae, Vibrio fluvialis, Vibrio furnissii, Vibrio harveyi, Vibrio parahaemolyticus and Vibrio vulnificus. Extracellular substances obtained from the cultural supernatant of MB I-3 by ethyl acetate extraction showed inhibitory effects on L. anguillarum. The antibacterial substance of MB I-3 was evaluated to destroy the cell membrane of L. anguillarum in electron micrographs. The probiotic effects of MB I-3 was tested by exposing olive flounder (Paralichthys olivaceus) fry to L. anguillarum with or without MB I-3. The cumulative mortality of olive flounder fry infected with L. anguillarum was 24% in the group with MB I-3, while it was 46% in the control group without MB I-3. These results indicate that MB I-3 has potential applications as a probiotic for the control of fish pathogenic vibrios in fish rearing system.

질병 치료를 위하여 사용되는 항균제는 효과적이고 사용이 편리한 장점이 있지만, 내성균 발생이나 항균물질 잔류와 같은 문제점들을 안고 있다. 따라서 어류 질병 치료 및 예방을 위해 어류와 인체에 안전한 생약제 개발과 함께 어류 질병 원인균의 과다 발생을 억제하며 어체의 건강을 유지시키기 위하여 유용 균주를 이용하고자 하는 연구가 다양하게 진행되고 있다. 본 연구는 이전 연구에서 분리되어 Listonella anguillarum에 대하여 생장 억제효과가 있는 것으로 밝혀진 Pseudomonas aeruginosa MB I-3 (MB I-3)를 이용하여 넙치의 비브리오병에 대한 생물학적 방제효과를 검토하였다. Double layered plate assay와 co-culture을 통하여 MB I-3의 L. anguillarum에 대한 생장 억제능력을 조사하였고, MB I-3 균주 배양액을 ethyl acetate로 추출하여 disk 확산법으로 추출물의 항균 효과를 확인하였다. 액체 및 고체 배양에서 생장이 억제된 L. anguillarum을 전자현미경으로 관찰하였으며, 넙치 치어 사육 수조에 L. anguillarum과 MB I-3 균주를 동시에 첨가하여 폐사율을 비교하였다. MB I-3는 8종의 병원성 비브리오균에 대하여 항균력을 나타내었으며, 96시간 동안 실시한 co-culture에서 L. anguillarum은 배양 후 9시간까지 생장 증가를 보였으나, 그 후 감소하는 경향을 보였다. 한편 MB I-3 배양액 추출물 또한 L. anguillarum에 항균활성을 보여 항균 물질이 ethyl acetate로 추출됨을 알 수 있었다. 전자현미경 관찰에서 L. anguillarum은 세포질의 밀도 감소 및 세포막의 swelling에 의한 세포 용해 현상을 보였다. 한편 MB I-3를 L. anguillarum과 함께 투여한 넙치 치어는 대조군에 비해 누적 폐사율이 약 20% 감소되는 결과를 보였으므로, MB I-3를 이용한 수산용 probiotics 개발 가능성을 시사하였다.

Keywords

References

  1. Austin, B. and Austin, D.A.: Bacterial fish pathogens, pp357-411, 5th ed., Springer Dordrecht Heiderberg, London UK, 2012.
  2. Balcazar, J.L., de Blas, I., Ruiz-Zarzuela, I., Cunningham, D., Vendrell, D. and Muzquiz, J.L.: The role of probiotics in aquaculture. Veterinary Microbiology, 114: 173-186, 2006. https://doi.org/10.1016/j.vetmic.2006.01.009
  3. Balcazar, J.L., Vendrell, D., de Blas, I., Ruiz-Zarzuela, I., Muzquiz, J.L. and Girones, O.: Characterization of probiotic properties of lactic acid bacteria isolated from intestinal microbiota of fish. Aquaculture, 278: 188-191, 2008. https://doi.org/10.1016/j.aquaculture.2008.03.014
  4. Byon, J. and Kim, E. : Inhibitory activity of marine bacterium on the growth of Vibrio anguillarum. J. Fish Pathol., 13: 1-6, 2000.
  5. Castillo, C.S., Wahid, M.I., Yoshikawa, T. and Sakata, T.; Isolation and inhibitory effect of anti-Vibrio substances from Pseudoalteromonas sp.A1-J11 isolated from the coastal sea water of Kagoshima bay. Fisheries Science, 78: 174-179, 2008.
  6. Cha, J.H., Yang, S.Y., Woo, S.H., Song, J.W., Oh, D.H. and Lee, K.J. : Effects of dietary supplementation with Bacillus sp. on growth performance, feed utilization, innate immunity and disease resistance against Streptococcus iniae in olive flounder Paralichthys olivaceus. Kor J Fish Aquat Sci, 45: 35-42, 2012. https://doi.org/10.5657/KFAS.2012.0035
  7. Chabrillon, M., Arijo, S., Diaz-Rsales, P., Balebona, M.C. and Morinigo, M.A.: Interference of Listonella anguillarum with potential probiotic microorganisms isolated from farmed gilthead seabream(Sparus aurata, L). Aquaculture Research, 37: 78-86, 2006. https://doi.org/10.1111/j.1365-2109.2005.01400.x
  8. Crab, R., Defoirdt, T., Bossier, P. and Verstraete, W.; Biofloc technology in aquaculture: Beneficial effects and future challenges. Aquaculture, 356-357: 351-356, 2012. https://doi.org/10.1016/j.aquaculture.2012.04.046
  9. Eddy, S.D. and Jones, S.H.: Microbiology of summer flounder Paralichthys dentatus fingerling production at a marine fish hatchery. Aquaculture, 211: 2-28, 2002.
  10. Hansen, G.H. and Olafsen, J.A.: Bacterial Interactions in early stages of marine cold water fish. Microb. Ecol., 38: 1-26, 1999. https://doi.org/10.1007/s002489900158
  11. Harikrishnan, R., Balasundaram, C. and Heo, M.S.: Effect of probiotics enriched diet on Paralichthys olivaceus infected with lymphocystis disease virus(LCDV). Fish and Shellfish Immunology, 29 : 868-874, 2010. https://doi.org/10.1016/j.fsi.2010.07.031
  12. Irianto, A. and Austin, B.: Probiotics in aquaculture. Journal of Fish Disease, 25: 633-642, 2002. https://doi.org/10.1046/j.1365-2761.2002.00422.x
  13. Kesarcodi-Watson, A., Kaspar, H., Lategan, M.J., Gibson, L.: Probiotics in aquaculture: The need, principles and mechanisms of action and screening processes. Aquaculture, 274: 1-14, 2008. https://doi.org/10.1016/j.aquaculture.2007.11.019
  14. Lategan, M.J., Booth, W., Shimmon, R. and Gibson, L.F. : An inhibitory substance produced Aeromonas media A199, an aquatic probiotic. Aquaculture, 254: 115-124, 2006. https://doi.org/10.1016/j.aquaculture.2005.11.019
  15. Moriarty, D.J.W.: The role of microorganisms in aquaculture ponds. Aquaculture, 151: 333-349, 1997. https://doi.org/10.1016/S0044-8486(96)01487-1
  16. Myouga, H., Yoshimizu, M., Tajima, K. and Ezura, Y.: Purification of an antiviral substance produced by Alteromonas sp. and its virucidal activity against fish viruses. Fish Pathology, 30: 15-22, 1995 https://doi.org/10.3147/jsfp.30.15
  17. Nakamura, A., Takahashi, K.G. and Katsuyoshi, M.: Vibriostatic bacteria isolated from rearing seawater of oyster brood stock : Potentiality as biocontrol agents for vibriosis in oyster larvae. Fish Pathology, 34: 139-144, 1999. https://doi.org/10.3147/jsfp.34.139
  18. Nwachi, O.F.: An overview of the importance of probiotics in aquaculture. J. fish. Aquat. Sci., 8: 30-32, 2013. https://doi.org/10.3923/jfas.2013.30.32
  19. Olafsen, J.A.: Interaction between fish larvae and bacteria in marine aquaculture. Aquaculture, 200: 223-247, 2001. https://doi.org/10.1016/S0044-8486(01)00702-5
  20. Ringo, E. and Birkbeck, T.H.: Intestinal microflora of fish larvae and fry. Aquaculture Research, 30: 73-93, 1999. https://doi.org/10.1046/j.1365-2109.1999.00302.x
  21. Riquelme, C., Araya, R. and Escribano, R.: Selective incorporation of bacteria by Argopecten purpuratus larvae: implications for the use of probiotics in culturing system of the Chilean scallop. Aquaculture, 181: 25-36, 2000. https://doi.org/10.1016/S0044-8486(99)00225-2
  22. Sugita, H., Shibuya, K., Shimooka, H. and Deguchi, Y.: Antibacterial abilities of intestinal bacteria in freshwater cultured fish. Aquaculture, 145: 195-203, 1996. https://doi.org/10.1016/S0044-8486(96)01319-1
  23. Tuan, T.N., Duc, P.M. and Hatai, K.: Overview of the use of probiotics in aquaculture. International Journal of Research in Fisheries and Aquaculture, 3: 89-97, 2013.
  24. Vine, N.G., Leukes, W.D., Kaiser, H., Daya, S., Baxter, J. and Hecht, T.: Competition for attachment of aquaculture candidate probiotic and pathogenic bacteria on fish intestinal mucus. Journal of Fish Disease, 27: 319-326, 2004. https://doi.org/10.1111/j.1365-2761.2004.00542.x
  25. Yang, B.G., Jun, Y.J. and Heo, M.S. : Screening and characterization of probiotic strains for prevention of bacterial fish disease. Kor. J. Microbiol. Biotechnol., 31: 129-134, 2003.
  26. 방종득, 이주석, 지보영, 김이청, 정승희, 심두생, 이윤호: 정균물질과 어병세균과의 상호작용 연구. 수진사업보고서: 254-270, 1999.