Taxonomy of Four Marine Bacterial Strains That Produce Tetrodotoxin

Abstract

Four strains of tetrodotoxin-producing bacteria isolated from a red alga and from pufferfish were characterized. Two of these strains are members of the genus Listonella MacDonell and Colwell. The phenotypic characteristics, guanine-plus-cytosine contents, and base sequences of the 16s rRNAs of these organisms indicated that they are members of ListoneUa pelagia (Vibrio pelagius) biovar II. The other two strains are members of the genus Alteromonas Baumann et al. and the genus Shewanella MacDonell and ColweU. These two strains are mutually distinct and distinct from the previously described Alteromonas and Shewanella species and therefore are placed in new species. The names Shewanella alga and Alteromonas tetraodonis are proposed for these organisms; the type strains are strains OK-1 and GFC, respectively.Tetrodotoxin, which is one of the strongest neurotoxins known (9, has been isolated from various species of animals, mostly animals which live in marine habitats (11,17). Although these animals themselves have long been considered the producers of tetrodotoxin, the recent isolation of tetrodotoxin-producing marine bacteria from various sources (12,17,18) suggests that the toxin is in fact produced by bacteria that are associated with the animals and their food. A presumptive Pseudomonas strain that was isolated from a red clacareous alga (Jania sp.) was reported to produce tetrodotoxin by Yasumoto et al. (18). Yasumoto et al. also reported the isolation of three other toxin-producing strains, one from the red alga and the other two from the surface slime of a pufferfish (Fugu poecilonotus) (19). In this paper we characterize and classify these four strains. Kotaki et al. (8) isolated strains OK-lT (T = type strain) and OK-2 from the surface of a red alga (Jania sp.) by using a medium containing 0.5% peptone (Nissui Seiyaku, Ltd.), 0.25% yeast extract (Nissui Seiyaku, Ltd.), 0.1% glucose, 3.0% NaC1, and 1.5% agar. The other two strains (strains GFB and GFCT) were isolated by Yasumoto and Yotsu from the skin slime of a pufferfish, using a similar medium (19). Along with strains OK-lT and GFCT, Alteromonas haloplanktis ATCC 14393T, Alteromonas nigrificans IAM 13010T, Pseudornonas nautica IAM 12929T, Deleya marina IAM 12928=, and Shewanella putrefaciens IAM 12079T were used for the analysis of 16s rRNA sequences. MATERIALS AND METHODS Strains.The strains were maintained in a semisolid agar medium (ORI medium) containing 0.1% Proteose Peptone no. 3 (Difco Laboratories, Detroit, Mich.), 0.1% yeast extract (Difco), 0.05% Phytone (BBL Microbiology Systems, Cockeysville, Md.), 0.02% sodium thiosulfate, 0.005% sodium sulfite, 0.04% ferric citrate, and 0.3% agar (Difco) in a mixture of aged seawater and distilled water (3: 1). The pH of the medium was adjusted to pH 7.6. Phenotypic characterization. A preliminary morphological and biochemical survey showed that two of the strains, strains OK-lT and GFCT, might belong to either the genus

Polyphasic taxonomic approach in the description of Alishewanella fetalis gen. nov., sp. nov., isolated from a human foetus.

“…The unidentified strain, CCUG 30811 T , the type strain of S. algae (IAM 14159 T ; Simidu et al, 1990), and the type strain of S. putrefaciens (ATCC 8071 T ; Derby & Hammer, 1931) were grown in veal infusion broth (VIB ; Difco), brain-heart infusion broth (BHI ; Oxoid), on Long & Hammer's medium (Van Spreekens, 1974), and on iron agar (Gram et al, 1987). Strains of S. putrefaciens and S. algae were grown at 25 and 37 mC, respectively.…”

Inference of interactions in cyanobacterial–heterotrophic co-cultures via transcriptome sequencing

“…PCC 7002 (hereafter Synechococcus 7002) in a co-culture with a marine facultative aerobe Shewanella putrefaciens W3-18-1 (hereafter Shewanella W3-18-1) under different trophic conditions. Although recognizing the opportunistic nature of interactions between these two strains, there is ample evidence that certain Shewanella species live in association with photoautotrophic prokaryotes (Simidu et al, 1990;Bowman et al, 1997;Salomon et al, 2003). Through the application of next-generation sequencing technology in conjunction with controlled cultivation and metabolomic profiling, we were able to delineate specific transcriptional responses of each organism to co-cultivation.…”

Shewanella amazonensis sp. nov., a novel metal-reducing facultative anaerobe from Amazonian shelf muds

“…Samples from the seasonally mobile intertidal deposits at the initiation of this coastal system (Jensen et al, 1980) and Shewanella benthica (MacDonnell & Colwell, 1985). Simidu et al (1990) …”