Systematic and Applied Microbiology 41 (2018) 340–347 Contents lists available at ScienceDirect Systematic and Applied Microbiology j ourna l ho mepage: www.elsevier.de/syapm Pseudomonas gallaeciensis sp. nov., isolated from crude-oil-contaminated intertidal sand samples after the Prestige oil spill Magdalena Mulet a , David Sánchez a , Ana C. Rodríguez a , Balbina Nogales a,b , Rafael Bosch a,b , Antonio Busquets a , Margarita Gomila a , Jorge Lalucat a,b , Elena García-Valdés a,b,∗ a Microbiologia, Departament de Biologia, Edifici Guillem Colom, Universitat de les Illes Balears, Campus UIB, 07122 Palma de Mallorca, Spain b Institut Mediterrani d’Estudis Avanc ¸ ats (IMEDEA, CSIC-UIB), Campus UIB, 07122 Palma de Mallorca, Spain a r t i c l e i n f o Article history: Received 5 July 2017 Received in revised form 26 March 2018 Accepted 30 March 2018 Keywords: Pseudomonas Pseudomonas pertucinogena group Contaminated sand Oil spill Genome a b s t r a c t Strains V113 T , V92 and V120 have been isolated from sand samples taken at the Atlantic intertidal shore in Galicia, Spain, after the Prestige oil spill. A preliminary analysis of the 16S rRNA and the partial rpoD gene sequences indicated that these strains belonged to the Pseudomonas genus, but they were distinct from any known Pseudomonas species. They were extensively characterized by a polyphasic taxonomic approach and phylogenetic data that confirmed that these strains belonged to the Pseudomonas pertu- cinogena group. Phylogenetic analysis of 16S rRNA, gyrB and rpoD gene sequences showed that the three strains were 99% similar and were closely related to members of the P. pertucinogena group, with less than 94% similarity to strains of established species; Pseudomonas pachastrellae was the closest relative. The Average Nucleotide Index based on blast values was 89.0% between V113 T and the P. pachastrellae type strain, below the accepted species level (95%). The predominant cellular fatty acid contents and whole cell protein profiles determined by MALDI-TOF mass spectrometry also differentiated the studied strains from known Pseudomonas species. We therefore conclude that strains V113 T , V92 and V120 represent a novel species of Pseudomonas, for which the name Pseudomonas gallaeciensis is proposed; the type strain is V113 T (= CCUG 67583 T = LMG 29038 T ). © 2018 Elsevier GmbH. All rights reserved. Introduction The genus Pseudomonas represents a group of Gram-negative bacteria within the Gammaproteobacteria that are non-spore form- ing rods that are motile by polar flagella [29]. The bacteria of the genus Pseudomonas are ubiquitous, metabolically versatile, and important for the recycling of elements in the environment. The genus was first described by Migula (1894) and currently comprises a large number of species, and new species are described contin- uously. At the time of this manuscript’s composition, 167 species with valid names have been described (http://www.bacterio.cict. fr/p/pseudomonas.html) [8,30]. ∗ Corresponding author at: Microbiologia, Departament de Biologia, Edifici Guillem Colom. Universitat de les Illes Balears, Crtra. Valldemosa Km 7.5, Campus UIB, 07122 Palma de Mallorca, Spain. E-mail address: elena.garciavaldes@uib.es (E. García-Valdés). The characterization of Pseudomonas type strains by multilo- cus sequence analysis (MLSA), concatenating the sequences of the 16S rDNA, gyrB, rpoD and rpoB genes, permitted the establishment of two main lineages, Pseudomonas fluorescens and Pseudomonas aeruginosa, which were divided into several groups (G) and sub- groups (SG) [25,27]. In recent years, the Pseudomonas pertucinogena G has experienced a significant increase in the number of newly described species compared with the other groups. In 2004, this group was represented only by P. pertucinogena [16]; today, it contains 16 species, the following of which have been recently described: Pseudomonas aestusnigri [34,11], Pseudomonas salegens [3], Pseudomonas salina [39], Pseudomonas oceani [36,10], Pseu- domonas populi [4] and ‘Pseudomonas saudimassiliensis’ not yet validated [6]. Proteobacteria have played a key role in the bacterial community in the contaminated intertidal sand after the Prestige oil spill [1,2]. The characterization of the bacterial species present in this habitat is crucial to have a comprehensive knowledge of the microorgan- isms present in this environment. This paper is focused on the https://doi.org/10.1016/j.syapm.2018.03.008 0723-2020/© 2018 Elsevier GmbH. All rights reserved.