Downloaded from www.microbiologyresearch.org by IP: 54.191.40.80 On: Fri, 08 Sep 2017 14:59:40 Vibrio renipiscarius sp. nov., isolated from cultured gilthead sea bream (Sparus aurata) Eva Tarazona, Marı ´a A. Ruvira, Teresa Lucena, M. Carmen Macia ´n, David R. Arahal and Marı ´a J. Pujalte Correspondence Marı ´a J. Pujalte maria.j.pujalte@uv.es Departamento de Microbiologı ´a y Ecologı ´a and Coleccio ´ n Espan ˜ ola de Cultivos Tipo (CECT), Universidad de Valencia, Valencia, Spain Two strains of Gram-negative, facultatively anaerobic, slightly halophilic bacteria, isolated from healthy gilthead sea bream (Sparus aurata) cultured in Spanish Mediterranean fish farms, were different from their closest relatives, Vibrio scophthalmi and V. ichthyoenteri, by phenotypic, phylogenetic and genomic standards. The strains were negative for decarboxylase tests and lacked extracellular hydrolytic activities, but were able to ferment D-mannitol, sucrose, cellobiose and D-gluconate, among other carbohydrates. The major cellular fatty acids were C 16:1 and C 16:0 , in agreement with other species of the genus Vibrio. Their 16S rRNA gene sequences were 98.4 and 97.2 % similar to those of the type strains of V. scophthalmi and V. ichthyoenteri, and the similarities using other housekeeping genes ( ftsZ, rpoD, recA, mreB and gyrB) and indices of genomic resemblance (average nucleotide identity and estimated DNA– DNA hybridization) between the isolates and those type strains were clearly below intraspecific levels, supporting the recognition of the strains as members of a separate novel species. Thus, we propose the name Vibrio renipiscarius sp. nov., with DCR 1-4-2 T (5CECT 8603 T 5KCTC 42287 T ) as the type strain. Vibrio, the most diverse genus of facultatively anaerobic, marine proteobacteria, currently contains more than 100 species (http://www.bacterio.net/vibrio.html). Aside from their prevalence among isolates obtained from marine water and animals, an important source of novel species of the genus Vibrio is the aquaculture environment: isolates obtained from cultured fish and shellfish around the world have provided more than 30 % of the species currently recognized in the genus. Since 2007, it has been usual (informally) to distribute species of the genus Vibrio in distinct lineages (clades) according to multilocus sequence analysis (MLSA)-defined phylogenetic relationships (Sawabe et al., 2007). Nineteen of these clades were reported in the recent study of Sawabe et al. (2013), some of them containing up to 17 species, others being much smaller. One of these defined clades is the so-called Scophthalmi clade, containing the species Vibrio scophthalmi (Cerda `-Cue ´llar et al., 1997) and Vibrio ichthyoenteri (Ishimaru et al., 1996), both isolated from the intestinal tracts of flatfish. The latter species is the agent of intestinal necrosis of flounder larvae (INFL), while the former is considered as non-pathogenic. Recently, Sawabe et al. (2013) have added another species to this clade, Vibrio ponticus (Macia ´n et al., 2004). During an MLSA study based on five housekeeping genes, undertaken on isolates of species of the genus Vibrio from aquaculture facilities and performed in our laboratory, an independent lineage within the Scophthalmi clade of the genus Vibrio was revealed (Tarazona et al., 2015). The present report describes the two strains that form this lineage as members of a novel species of the genus Vibrio named Vibrio renipiscarius sp. nov. Strains DCR 1-4-2 T and DCR 1-4-12 were isolated in Feb- ruary 2000 from the head kidney of gilthead sea bream (Sparus aurata) cultivated in a fish farm on the Spanish Mediterranean coast. Isolation was achieved by direct plating of a loop of head kidney on marine agar 2216 (MA) and tryptone soy agar plus 2 % NaCl, incubated at 26 8C, in the Instituto de Acuicultura Torre de la Sal (IATS-CSIC) (Castello ´ n, Spain). The strains were charac- terized phenotypically through a selection of tests and Abbreviations: ANI, average nucleotide identity; DDH, DNA–DNA hybridization; MLSA, multilocus sequence analysis. The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA, recA, gyrB, ftsZ, rpoD and mreB gene sequences of strain DRC 1-4-2 T are HG931128, LN626597, LN626503, LN626550, LN626455 and LN626407. For DCR 1-4-12, the corresponding accession numbers are HG931126, LN626596, LN626502, LN626549, LN626454 and LN626406. The accession numbers for genome sequence data for strains DCR 1-4-2 T and DCR 1-4-12 are JTKH00000000 and JTKI00000000, respectively. A supplementary table and a supplementary figure are available in the online Supplementary Material. International Journal of Systematic and Evolutionary Microbiology (2015), 65, 1941–1945 DOI 10.1099/ijs.0.000200 000200 G 2015 IUMS Printed in Great Britain 1941