바로가기메뉴

본문 바로가기 주메뉴 바로가기

ACOMS+ 및 학술지 리포지터리 설명회

  • 한국과학기술정보연구원(KISTI) 서울분원 대회의실(별관 3층)
  • 2024년 07월 03일(수) 13:30
 

logo

메뉴

Characterization and species-specific detection of Vibrio parahaemolyticus-related species associated with mass mortality of ark shell (Scapharca broughtonii)

Abstract

Ark shell (Scapharca broughtonii) is cultured in southern coast of Korea and is a commercially important shell in aquaculture. However its productivity has decreased rapidly in the last decade due to mass mortality. Various microbial investigations were performed to identify the caused bacterial species for economic losses. Using cumulative mortality analysis, 16S rDNA sequence analysis, and API 20 kit assay, Vibrio parahaemolyticus-related species (8M 4-1) was identified as an etiologic bacterium. The LD50 value of the isolated bacterium to ark shell was lower than 106 CFU/mL. After infection, histological diagnosis from hepatopancreatic tissue also provided evidence for pathogenicity. To monitor this ark shell pathogenic bacterium, we designed the PCR detection method with the Vibrio species groEL gene. We also tested various virulence gene-targeted detection methods, but amplified products were not suitable for V. parahaemolyticus recognition. In this study, we identified V. parahaemolyticusrelated species as a causative organism of mass mortality, and demonstrated that PCR detection assay could successfully detect V. parahaemolyticus strains from infected tissues with high sensitivity. Using these results, we identified V. parahaemolyticus-related 8M 4-1 species as the causative agent of ark shell mortality and showed that newly developed PCR detection method was useful for pathogen monitoring.

keywords
Ark shell (Scapharca broughtonii), mass mortality, Vibrio parahaemolyticus, PCR

참고문헌

1.

Afi, A., Matthieu, G., Françoise, A, Roger, K., Xavier, P., Jean-Louis, N., (2010) Occurence and seasonality of Vibiro aestuarianus in sediment and Crassostrea gigas haemolymph at two oyster farms in France. Disease of Aquatic Organisms, 91: 213-221.

2.

Bej, A.K., Patterson, D.P., Brasher, C.W., Vickery, MC., Jones, D.D., Kaysner, C.A., (1999) Detection of total and hemolysin-producing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tl, tdh and trh. Journal of Microbiological Method, 36:215-225.

3.

Cabral, J.P.S., (2010) Water Microbiology. Bacterial pathogens and water. International Journal of Environmental Research and Public Health, 7:3657-3703.

4.

Cai, J.P., Han, Y., Wang, Z., (2006) Isolation of Vibrio parahaemolyticus from abalone (Haliotis diversicolor supertexta L.) postlarvae associated with mass mortalities. Aquaculture, 257: 161-166.

5.

Chatterjee, S., Halder, S., (2012) Vibrio related diseases in aquaculture and development of rapid and accurate identification methods. Journal of Marine Science: Research & Development, http://dx.doi.org/10.4172/2155-9910.S1-002.

6.

Cheng, L., Huang, J., Ahi, C., Thompson, K., Mackey, B., Cai, J., (2008) Vibrio parahaemolyticus associated with mass mortality of postlarval alblone, Haliotis diversicolor supertexta (L.), in Sanya, China. Journal of the World Aquaculture Society, 39: 746-757.

7.

Cheng, W., Hsiao, I.S., Hsuc, C.H., Chen, J.C., (2004)Change in water temperature on the immune response of Taiwan abalone Haliotis diversicolor supertexta and its susceptibility to Vibrio parahaemolyticus. Fish and Shellfish Immunology, 17: 235-243.

8.

Chun, Y.Y., Na, G.H., Choi, W.J., (1991) Mass mortality of ark shell, Anadara broughtonii, SCHRENCK seeding with marine ecological characteristics. Journal of the Korean Fisheries Society, 24: 70-78. [in Korean with English abstract].

9.

Coutard, F., Pommepuy, M., Loaec, S., Hervio-Health, D., (2005) mRNA detection by reverse transcription-PCR for monitoring viability and potential virulence in a pathogenic strain of Vibrio parahaemolyticus in viable but nonculturable state. Journal of Applied Microbiology, 98: 951-961.

10.

Feng, J.W., Wang, X.Y., Ki D.L., Tang, S.M., You, S.Z., (2007) Study on rapid detection of 2 kinds of Vibrio spp. In seafood by real-time fluorescence PCR. Chinese Journal of Food Hygiene, 19: 514-517. [in Chinese].

11.

Frans, I., Michiels, C.W., Bossier, P., Willems, K.A., Lievens, B., Rediers, H., (2011) Vibrio anguillarum as a fish pathogen: virulence factors, diagnosis and prevention. Journal of Fish Diseases, 34: 643-661.

12.

González, S.F., Krug, M.J., Nielsen, M.E., Santos, Y., Call, D.R., (2004) Simultaneous detection of marine fish pathogens by using multiplex PCR and a DNA microarray. Journal of Clinical Microbiology, 42:1414-1419.

13.

Gwon B.G., Kim Y.O., Nam B.H., Kim W.J., Kong H.J., Kim B.S., Jee Y.J., Lee S.J., An C.M., Kim D.G., (2013) Analysis of diversity of hemolytic microbiome from aquafarm of arkfarm, Scapharca broughtonii. Journal of Fish Pathology, 26: 193-206.

14.

Hoff K.J., Tech M., Lingner T., Daniel R., Morgenstern B., Meinicke P., (2008) Gene prediction in metagenomic fragments: a large scale machine learning approach. BMC Bioinformatics, 9: 217-230.

15.

Hong, G.E, Kim D.G., Bae, J.Y., Ahn, S.H., Bai, S.C., Kong, I.S., (2007) Species-specific PCR detection of the fish pathogen, Vibrio anguillarum, using the amiB gene, which encodes N-acetylmuramoyl-L-alanine amidase. FEMS Microbiology Letters, 269:201-206.

16.

Hossain, M.T., Kim, E.Y., Kim, Y.R., Kim, D.G., Kong, I.S., (2011) Application of groEL gene for the species-specific detection of Vibrio parahaemolyticus by PCR. Letters in Applied Microbiology, 54: 67-72.

17.

Jo, G.A., Kwon, S.B., Kim, N.K., Hossain, M.T., Kim, Y.R., Kim, E.Y., Kong, I.S., (2013) Species-specific duplex PCR for detecting the important fish pathogens Vibrio anguillarum and Edwardsiella tarda. Fisheries and Aquatic Sciences, 16: 273-277.

18.

Kim, B.H., Shin, Y.K., Park, K.Y., Choi, N.J., Oh, B.S., Min, B.H., (2008) Growth and survival of the spat of ark shell, Scapharca broughtonii in intermediate culture with different shape of protective net od spat loss. Korean Journal of Malacology, 24: 131-136. [in Korean with English abstract].

19.

Kim, D.G., Bae, J.Y., Hong, G.E., Min, M.K., Kim, J.K, Kong, I.S., (2008) Application of the rpoS gene for the detection of Vibrio anguillarum in flounder and prawn by polymerase chain reaction. Journal of Fish Diseases, 31: 639-647.

20.

Kim, D.G., Nam, B.H., Kong, H.J., Kim, W.J., Kim, B.S., Jee, Y.J., Lee, S.J., Jung, C.G., Kong, M.S., Kim, Y.O., (2012) Analysis of hemolytic microflora from the ark shell (Scapharca broughtonii). Journal of Life Science, 22: 642-649. [in Korean with English abstract].

21.

Kim, Y.B., Okuda, J., Matsumoto, C., Takahashi, N., Hashimoto, S., Nishibuchi, M., (1999) Identification of Vibrio parahaemolyticus strains at the species level by PCR targeted to the toxR gene. Journal of Clinical Microbiology, 37: 1173-1177.

22.

Koh, B.H., Lee, W.D., Ann, S.K., Kim, J.H., Lee, M.S., (1997) Effect of storage temperature on the survival of Vibrio mimicus K-1 in seawater and ark shell. Journal of Korean Fisheries Society, 30: 277-281. [in Korean with English abstract].

23.

Lacoste, A., Jalabert, F., Malham, S., Cueff, A., Gelebart, F., Cordevant, C., Lange, M., Poulet, S., (2001) A Vibrio splendidus strain is associated with summer mortality of juvenile oysters Crassostrae gigias in the Bay of Morlaix. Diseases of Aquatic Organisms, 46: 139-145.

24.

Liu, P.C., Chen, Y.C., Huang, C.Y., Lee, K.K., (2000)Virulence of Vibrio parahaemolyticus isolated from cultured small abalone Haliotis diversicolor supertexta, with withering syndrome. Letters in Applied Microbiology, 31: 433-437.

25.

Manns, J.M., Mosser, D.M., Buckley, H.R., (1994)Production of a hemolytic factor by Candida albicans. Infection and Immunity, 62: 5154-5165.

26.

Paillard, C., Roux, L.F., Borrego, J.J., (2004) Bacterial disease in marine bivalves, a review of recent studies: Trends and evolution. Aquatic Living Resources, 17: 477-498.

27.

Park, M.S., Lim, H.J., Kim, P.J., (1998) Effect of environmental factors on the growth, glycogen and hemoglobin content of cultured arkshell, Scapharca broughtonii. Journal of Korean Fisheries Society, 31:176-185. [in Korean with English abstract].

28.

Park, S.I., (2009) Disease control in Korea aquaculture. Fish Pathology, 44: 19-23.

29.

Ruangpan, L., Kitao, T., (1991) Vibrio bacteria isolated from black tiger shrimp, Penaeus mondon Fabricius. Journal of Fish Diseases, 14: 383-388.

30.

Schmidt, T.M., DeLong, E.F., Pace, N.R., (1991) Analysis of a marine picoplankton community by 16S rDNA gene cloning and sequence. Journal of Bacteriology, 173: 4371-4378.

31.

Shin, Y.K., Kim, B.H., Choi, N.J., Jung, C.G., Park, M.W., (2008) Influence of temperature, salinity and hypoxia on survival and metabolic rate in the ark shell Scapharca broughtonii. Korean Journal of Malacollogy, 24: 59-65. [in Korean with English abstract].

32.

Sorokin, V.A., Gelfand, M.S., Artamonova, I.I., (2010)Evolutionary dynamics of clustered irregularly interspaced short palindromic repeat systems in the ocean metagenome. Applied and Environmental Microbiology, 76: 2136-2144.

33.

Suthienkul, O., Ishibashi, M., Iida, T., Nettip, N., Supavej, S., Eampokalap, B., Makino, M., Honda, T., (1995) Urease production correlates with possession of the trh gene in Vibrio parahaemolyticus strains isolated in Thailand. The Journal of Infectious Diseases, 172: 1405-1408.

34.

Suzuki, M.T., Rappé, M.S., Haimberger, Z.W., Winfield, H., Adair, N., Ströbel, J., Giovannoni, S.J., (1997)Bacterial diversity among small-subunit rRNA gene cloned and cellular isolates from the same seawater sample. Applied and Environmental Microbiology, 63:983-989.

35.

Torkildsen, L., Lambert, C., Nylund, A., Magnesen, T., Bergh, O., (2005) Bacteria associated with early life stages of the great scallop, Pecten maximus: impact on larval survival. Aquaculture International, 13:575-592.

36.

Travers, M.A., Le, G.N., Huchette, s., Koken, M., Pillard, C., (2008) Summer immune depression associated with increased susceptibility of the European abalone, Haltiotis tuberculata to Vibrio harveyi infection. Fish and Shellfish Immunology, 25:800-808.

37.

Woo, P.C., Ng, K.H., Lau, S.K., Yip, K.T., Fung, A.M. Leung, K.W., Tam, D.M., Que, T.L., Yuen, K.Y., (2003) Usefulness of the MicroSeq 500 16S ribosomal DNA-based significant bacterial isolates with ambiguous biochemical profiles. Journal of Clinical Microbiology, 41: 1996-2001.

38.

Yoo, S.K. (1986) Coastal culture. fifth ed. Gudeok publisher. Busan.

39.

Yue, X., Liu, B., Xiang, J., Jia, J., (2010) Identification and characterization of the pathogenic effect of a Vibrio parahaemolytics-related bacterium isolated from clam Meretrix meretrix with mass mortality. Journal of Invertebrate Pathology, 103: 109-115.

logo