Shewanella marinintestina sp. nov., Shewanella schlegeliana sp. nov. and Shewanella sairae sp. nov., novel eicosapentaenoic-acid-producing marine bacteria isolated from sea-animal intestines.

Abstract

Three novel Shewanella species are described on the basis of phenotypic, chemotaxonomic and phylogenetic studies. A total of six novel halophilic, aerobic organisms with the ability to produce eicosapentaenoic acid (EPA) were isolated from various sea animals in Japan. Cells of all six isolates were Gram-negative, rod-shaped and motile by means of polar flagella. They were able to produce large amounts of EPA (about 20% of the total fatty acids) and had isoprenoid quinones Q-7 and Q-8 as major components. Analysis of the nearly complete 16S rRNA gene sequences of the novel isolates showed that they are very close phylogenetically (sequence similarity > 99%) and the closest species was Shewanella pealeana, with 97% sequence similarity. However, analysis of gyrB sequences indicated that the novel isolates were divided into three groups at sufficient phylogenetic distance to indicate that they are different species (< 90% sequence similarity). DNA-DNA hybridization experiments supported this conclusion. The first group (three strains) had positive reactions for lipase, DNase, ONPG and trimethylamine oxide (TMAO) reduction and had G + C contents of 43 mol% (determined by HPLC). The second group (two strains) was positive for urease, DNase, ONPG and TMAO reduction but not lipase. Their G + C content was 45 mol%. The third group (one strain) was negative for ONPG, DNase and TMAO reduction and had a G + C content of 43 mol%. Strains of the second group, but not those of the first or third groups, grew at 32 degrees C. On the basis of the polyphasic taxonomic data, the novel strains isolated from intestines of sea animals are placed in three novel species of the genus Shewanella: Shewanella marinintestina sp. nov. (type strain: JCM 11558T =LMG 21403T), Shewanella schlegeliana sp. nov. (type strain: JCM 11561T =LMG 21406T) and Shewanella sairae sp. nov. (type strain: JCM 11563T =LMG 21408T).