Downloaded from www.microbiologyresearch.org by IP: 54.157.13.203 On: Sun, 07 Feb 2016 10:22:48 Tenacibaculum soleae sp. nov., isolated from diseased sole (Solea senegalensis Kaup) Maximino Pin ˜ eiro-Vidal, 1 Cristina G. Carballas, 1 Oscar Go ´ mez-Barreiro, 2 Ana Riaza 2 and Ysabel Santos 1 Correspondence Ysabel Santos ysantos@usc.es 1 Departamento de Microbiologı ´a y Parasitologı ´a, Edificio CIBUS, Facultad de Biologı ´a, Universidad de Santiago de Compostela, Campus Sur, 15782 Santiago de Compostela, Spain 2 Stoltseafarm S.A., Lira, Spain A novel Gram-negative, rod-shaped, gliding bacterial strain designated LL04 12.1.7 T was isolated from diseased sole (Solea senegalensis Kaup) in Galicia, Spain. Colonies were yellow-pigmented with uneven edges and did not adhere to the agar. The DNA G+C content of the strain was 29.8 mol%. 16S rRNA gene sequence similarity analysis indicated that strain LL04 12.1.7 T is a member of the genus Tenacibaculum in the family Flavobacteriaceae. Sequence similarities between the isolate and the type strains of other members of the genus were 96.7–94.8 %. The major fatty acids (.10 % of total fatty acids) were iso-C 15 : 0 (23.1 %), iso-C 15 : 0 3-OH (10.6 %), C 15 : 1 v6c (12.2 %) and summed feature 3 (comprising C 16 : 1 v7c and/or iso-C 15 : 0 2-OH, 11.0 %). Genotypic and phenotypic data distinguished strain LL04 12.1.7 T from the 11 recognized Tenacibaculum species, indicating that it represents a novel species, for which the name Tenacibaculum soleae sp. nov. is proposed. The type strain is strain LL04 12.1.7 T (5CECT 7292 T 5NCIMB 14368 T ). The genus Tenacibaculum (Suzuki et al., 2001) in the family Flavobacteriaceae (Reichenbach, 1992a, b; Bernardet et al., 1996, 2002) currently comprises 11 species derived from different marine ecosystems and intensive aqua- culture systems. Tenacibaculum maritimum and Tenacibaculum discolor were isolated from diseased fish (Wakabayashi et al., 1986; Pin ˜eiro-Vidal et al., 2008); Tenacibaculum ovolyticum was isolated from fish eggs (Hansen et al., 1992); Tenacibaculum litopenaei (Sheu et al., 2007) and Tenacibaculum gallaicum (Pin ˜eiro-Vidal et al., 2008) were from seawater of shrimp- and turbot-holding tanks, respectively; Tenacibaculum amylolyticum and Tenacibaculum mesophilum were from marine macroalgae and sponge, respectively (Suzuki et al., 2001); Tenacibaculum skagerrakense was from seawater (Frette et al., 2004); and Tenacibaculum lutimaris (Yoon et al., 2005), Tenacibaculum litoreum (Choi et al., 2006) and Tenacibaculum aestuarii (Jung et al., 2006) were from tidal flat sediment. During the characterization of bacteria isolated from a diseased cultured sole (Solea senegalensis Kaup), strain LL04 12.1.7 T was recovered on plates of Flexibacter maritimus medium (FMM) (Pazos et al., 1996). Subcultivation was done on FMM or marine agar 2216 (MA; Difco) at 25 u C for 48 h. Strains were preserved at 280 u C in both marine broth 2216 (MB; Difco) supple- mented with 15 % (v/v) glycerol and Microbank tubes (Prolab Diagnostics). Experimental infection assays have demonstrated that strain LL04 12.1.7 T is virulent for fingerlings of sole and turbot, but not for mice (data not shown). Morphological, physiological and biochemical tests were performed as described by Bernardet et al. (2002). The Gram reaction was tested by using the bioMe ´rieux Gram stain kit according to the manufacturer’s instructions and the non-staining KOH method (Buck, 1982). Gliding motility was determined by phase-contrast microscopic examination of a fresh MB culture by the hanging drop technique as recommended by Bernardet et al. (2002). The presence of flexirubin-type pigments was determined by using the KOH test as described by Reichenbach (1989). Catalase and oxidase activities were determined as described by Cowan & Steel (1965). The capacity of the strain to grow under anaerobic conditions was tested on MA using the GasPak anaerobic system (BBL). The optimal pH and the pH range for growth were determined in FMM broth adjusted to pH 4–10 according to Suzuki et al. (2001). Growth at various temperatures (8, 15, 18, 22, 25, 30, 37 and 44 u C) was determined on FMM agar plates. Tolerance to salinity was tested in FMM broth containing 10, 20, 30, 50, 70 or 100 % seawater or 0.8, 1, 3, 5, 7 or 10 % (w/v) NaCl. Indole and H 2 S production were tested on FMM broth supplemented with 1 % (w/v) tryptone or 5 % The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain LL04 12.1.7 T is AM746476. International Journal of Systematic and Evolutionary Microbiology (2008), 58, 881–885 DOI 10.1099/ijs.0.65539-0 65539 G 2008 IUMS Printed in Great Britain 881