Abstract
A bacterium S184 isolated from the liver of puffer fish was found to produce tetrodotoxin (TTX). The toxin was purified from the cultures by activated charcoal column and C18 cartridge column after being cultured at 28 °C in broth medium. The results of toxicity mouse bioassay, liquid chromatography/fluorescent detection (LC/FLD), liquid chromatography/mass spectrometry (LC/MS), liquid chromatography/tandem mass spectrometry (LC/MS/MS) suggested that the strain could produce TTX during cultivation. The strain was identified asBacillus horikoshii according to its physiological-biochemical characteristics and phylogenetic analysis based on 16S rDNA sequences.
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Asakawa M., Toyoshima T., Ito K., Yamaguchi C., Tsunetsugu S., Shida Y., Kajihara H., Mawatari S.F., Noguchi T., Miyazawa K. (2003). Paralytic toxicity in the ribbon wormCephalothrix species (Nemertea) in Hiroshima Bay, Hiroshima Prefecture, Japan and the isolateion of tetrodotoxin as a main component of its toxins. Toxicon, 41: 747–753.
Carroll S., McEvoy E.G., Gibson R. (2003). The production of tetrodotoxin-like substances by nemertean worms in conjunction with bacteria. J. Exp. Mar.Biol. Ecol., 288: 51–63.
Cheng C.A., Hwang D.F., Tsai Y.H., Chen H.C., Jeng S.S., Noguchi T., Ohwada K., Hasimoto K. (1995). Microflora and tetrodotoxin-producing bacteria in a gastropodNiotha clathrata. Fd. Chem. Toxic., 33: 929–934.
Claus D., Berkeley R.C.W. (1986). GenusBacillus Cohn 1872. In: Sneath P.H.A., Mair N.S., Sharpe M.E., Holt J.G., Eds., Bergey’s Manual of Systematic Bacteriology, vol. 2, Baltimore, Williams and Wilkins, pp. 1105–1139.
Daly J., Gusovsky F., Miyer C., Yotsu-Yamashita M., Yasumoto T. (1994). First occurrence of tetrodotoxin in a dendrobatid frog (Colostethus inguinalis), with further reports for the bufonid genusAtelopus. Toxicon, 32: 279–285.
Do H.K., Kogure K., Simidu U. (1990). Identification of deep-sea-sediment bacteria which produce tetrodotoxin. Appl. Environ. Microbiol., 56: 1162–1163.
Do H.K., Kogure K., Imada C., Noguchi T., Ohwada K., Simidu U. (1991). Tetrodotoxin production of actinomycetes isolated from marine sediment. J. Appl. Bacteriol., 70: 464–468.
Do H.K., Hamasaki K., Ohwada K., Simidu U., Noguchi T., Shida Y., Kogure K. (1993). Presence of tetrodotoxin and tetrodotoxin-producing bacteria in freshwater sediments. Appl. Environ. Microbiol., 59: 3934–3937.
Gordon R.E., Haynes W.C., Pang C.H.N. (1973). GenusBacillus. In: Agricultural Handbook Number 427, US Department of Agriculture, Washington, DC.
Hanifin C.T., Brodie III E.D., Brodie Jr. E.D. (2002). Tetrodotoxin levels of the rough-skin newt,Taricha granulose, increase in long-term captivity. Toxicon, 40: 1149–1153.
Hwang D.F., Arakawa O., Saito T., Noguchi T., Simidu U., Tsukamoto K., Shida Y., Hashimoto K. (1989). Tetrodotoxin-producing bacteria from the blue-ringed cotopusOctopus maculosus. Mar. Biol., 100: 327–332.
Hwang D.F., Chueh C.H., Jeng S.S. (1990). Occurrence of tetrodotoxin in the gastropods molluskNatica lineate (lined moon shell). Toxicon, 28: 21–27.
Lee M., Jeong D., Kim W., Kim W., Kim H., Kim C., Park W., Park Y., Kim K., Kim H., Kim D. (2000). A tetrodotoxin-producingVibrio strain, LM-1, from the puffer fishFugu vermicularis radiatus. Appl. Environ. Microbiol., 66: 1698–1701.
Lane D.J., Pace B., Olsen G.J., Stahl D.A., Sogin M.L., Pace N.R. (1985). Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses. Proc. Natl. Acad. Sci. USA., 82: 6955–6959.
Li Z., Kawamura Y., Shida O., Yamagata S., Deguchi T., Ezaki T. (2002).Bacillus okuhidensis sp. nov., isolated from the Okuhida spa ares of Japan. Int. J. Syst. Evol. Microbiol., 52: 1205–1209.
Logan N.A., Lebbe L., Verhelst A., Goris J., Forsyth G., Rodriguez-Diaz M., Heyndrickx M., De Vos. P., (2002).Bacillus luciferensis sp. nov., from volcanic soil on Candlemas Island, South Sandwich archipelago. Int. J. Syst. Evol. Microbiol., 52: 1985–1989.
Nielsen P., Fritze D., Priest F.G. (1995). Phenetic diversity of alkaliphilicBacillus strains: proposal for nine new species. Microbiology, 141: 1745–1761.
Noguchi T., Hashimoto Y. (1973). Isolation of tetrodotoxin from a gobyGobius criniger. Toxicon, 11: 305–307.
Noguchi T., Jeon J.K., Arakawa O., Sugita H., Deguchi Y., Shida Y., Hashimoto K. (1986). Occurrence of tetrodotoxin and anhydrotetrodotoxin inVibrio sp. isolated from the intestines of a Xanthid Crab,Atergatis floridus. J. Biochem., 99: 311–314.
Noguchi T., Hwang D.F., Arakawa O., Sugita H., Deguchi Y., Shida Y., Hashimoto K. (1987).Vibrio alginolyticus, a tetrodotoxin-producing bacterium, in the intestines of the fishFugu vermicularis vermicularis. Mar. Biol., 94: 625–630.
Ritchie K.M., Nagelkerken I., James S., Smith G.W. (2000). A tetrodotoxin-producing marine pathogen. Nature, 404: 354.
Thuesen E.V., Kogure K. (1989). Bacterial production of tetrodotoxin in four species of chaetognatha. Biol. Bull., 176: 191–194.
Tsai Y.H., Ho P.H., Jeng S.S., Hwang D.F. (2002). Paralytic toxins in Taiwanese crabXanthias lividus. Fish. Sci., 68: 659–661.
Tsai Y.H., Chao S.M., Lin G.T., Noguchi T., Hwang D.F.(2004). Tetrodotoxin of the starfishAstropecten vappa collected from western Taiwan. Fish. Sci., 70: 934–936.
Wu Z., Xie L., Xia G., Zhang J., Nie Y., Hu J., Wang S., Zhang R. (2005a). A new tetrodotoxin-producing actinomycete,Nocardiopsis dassonvillei, isolated from the ovaries of puffer fishFugu rubripes. Toxicon, 45: 851–859.
Wu Z., Yang Y., Xie L., Xia G., Hu J., Wang S., Zhang R. (2005b). Toxicity and distribution of tetrodotoxin-producing bacteria in puffer fishFugu rubripes collected from the Bohai sea of China. Toxicon, 46: 471–476.
Yan Q., Yu P.H., Li H. (2005). Detection of tetrodotoxin and bacterial production by Serratia marcescens. World J. Microbiol. Biotechnol., 21: 1255–1258.
Yasumoto T., Nagai H., Yasumura D., Michishita T., Endo A., Yotsu M., Kotaki Y. (1986a). Interspecies distribution and possible origin of tetrodotoxin. Ann. New. York. Acad. Sci., 479: 44–51.
Yasumoto T., Yasumura D., Yotsu M., Michishita T., Endo A., Kotaki Y. (1986b). Bacterial production of tetrodotoxin and anhydrotetrodotoxin. Agri. Biol. Chem., 50: 793–795.
Yu C.F., Yu P.H., Chan P.L., Yan Q., Wong P.K. (2004). Two novel species of tetrodotoxin-producing bacteria isolated from toxic marine puffer fishes. Toxicon, 44: 641–647.
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Lu, Y., Yi, R. Bacillus horikoshii, a tetrodotoxin-producing bacterium isolated from the liver of puffer fish. Ann. Microbiol. 59, 453–458 (2009). https://doi.org/10.1007/BF03175130
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DOI: https://doi.org/10.1007/BF03175130