Lutibacter Holmesii Sp. Nov., a Marine Bacterium of the Family

International Journal of Systematic and Evolutionary Microbiology (2015), 65, 3991–3996 DOI 10.1099/ijsem.0.000525

Lutibacter holmesii sp. nov., a marine bacterium of the family Flavobacteriaceae isolated from the sea urchin Strongylocentrotus intermedius, and emended description of the genus Lutibacter Olga I. Nedashkovskaya,1,2 Stefanie Van Trappen,3 Natalia V. Zhukova2,4 and Paul De Vos3

Correspondence 1G.B. Elyakov Paciﬁc Institute of Bioorganic Chemistry of the Far-Eastern Branch of the Russian Olga I. Nedashkovskaya Academy of Sciences, Prospekt 100 Let Vladivostoku 159, 690022 Vladivostok, Russia [email protected] 2Far Easten Federal University, Sukhanova St. 8, 690091 Vladivostok, Russia or 3BCCM/LMG Bacteria Collection, and Laboratory of Microbiology, Ghent University, Ledeganck- [email protected] straat 35, B-9000 Ghent, Belgium 4A.V. Zhirmunsky Institute of Marine Biology of the Far Eastern Branch of the Russian Academy of Science, Pal’chevskogo St. 17, 690032 Vladivostok, Russia

Seven Gram-staining-negative, strictly aerobic, pale-yellow-pigmented, rod-shaped and non- motile strains were isolated from the sea urchin Strongylocentrotus intermedius collected from Troitsa Bay, Sea of Japan. Phylogenetic analyses based on 16S rRNA gene sequences showed that these isolates were afﬁliated with the family Flavobacteriaceae. The novel isolates showed 99.9–100 % 16S rRNA gene sequence similarity to each other and were closely related to the type strains of the recognized members of the genus Lutibacter with sequence similarities of 95.8–98.4 %. The G+C content of the genomic DNA was 35–36 mol%. DNA–DNA relatedness among the sea urchin isolates was 95–99 % and between strain KMM 6277T and its most closely related type strains, Lutibacter agarilyticus KCTC 23842T and Lutibacter litoralis JCM T 13034 , was 38 and 27 %, respectively. The prevalent fatty acids were iso-C15 : 0, anteiso-C15 : 0,

summed feature 3 (comprising iso-C15 : 0 2-OH and/or C16 : 1 v7c fatty acids), iso-C15 : 1 and

C15 : 0. The polar lipid proﬁle was composed of the phosphatidylethanolamine, one unknown aminolipid and one unknown lipid. The main respiratory isoprenoid quinone was MK-6.The results of phylogenetic, phenotypic and genotypic analyses indicated that the novel strains represent a novel species within the genus Lutibacter, for which the name Lutibacter holmesii sp.nov.is proposed. The type strain is KMM 6277T (5 CCUG 62221T 5 LMG 26737T).

The genus Lutibacter was proposed to accommodate het- Choi et al., 2013). The type strains of Lutibacter agarilyticus erotrophic, aerobic, Gram-staining-negative, rod-shaped and Lutibacter oricola were isolated from shallow coastal and oxidase-negative marine bacteria within the family Fla- seawater (Park et al., 2013; Sung et al., 2015). In the vobacteriaceae (Choi & Cho, 2006). At the time of writing, course of a survey of bacterial diversity of the edible sea the genus Lutibacter comprises six species with validly pub- urchin Strongylocentrotus intermedius, seven heterotrophic, lished names. The type strain of the type species, Lutibacter aerobic, pale-yellow-pigmented strains were isolated. The litoralis, and those of Lutibacter maritimus, Lutibacter aes- results of phylogenetic analysis based on 16S rRNA gene tuarii and Lutibacter flavus were recovered from tidal flat sequencing revealed that the novel isolates belonged to sediments collected on the coast of the Korean peninsula the family Flavobacteriaceae and formed a distinct evol- (Choi & Cho, 2006; Park et al., 2010; Lee et al., 2012; utionary lineage within the genus Lutibacter. The present study aimed to report on the isolation and taxonomic identification of the novel strains using a polyphasic The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA approach. T gene sequences of Lutibacter holmesii KMM 6277 , KMM 6302, KMM T 6303, KMM 6305, KMM 6307, KMM 6308 and KMM 6311 are Strains KMM 6277 , KMM 6302, KMM 6303, KMM 6305, JQ241142, JQ241143, JQ241144, JQ241145, JQ241146, KMM 6307, KMM 6308 and KMM 6311 were isolated from JQ241147 and JQ241148, respectively. the sea urchin S. intermedius that was collected in Troitsa

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Bay, Gulf of Peter the Great, the Sea of Japan, as described closely related to L. agarilyticus KCTC 23842T and L. litor- previously (Nedashkovskaya et al., 2012). After primary alis JCM 13034T with 98.4 % and 97.3 % 16S rRNA gene isolation and puriﬁcation, strains were cultivated at 28 8C sequence similarity, respectively, and shared less than on marine agar (Difco) and stored at 280 8C in marine 97 % sequence similarity to the type strains of other recog- broth (Difco) supplemented with 20 % (v/v) glycerol. nized species of the genus Lutibacter (Fig. 1).

DNA extraction, PCR and 16S rRNA gene sequencing were DNA was isolated following the method of Marmur (1961) carried out as described by Vancanneyt et al. (2006). The and the DNA G+C content was determined by the thermal alignment of the 16S rRNA gene sequences obtained for denaturation method (Marmur & Doty, 1962). DNA–DNA the novel strains with those retrieved from GenBank, and hybridization experiments were performed spectrophotome- phylogenetic analysis and bootstrap analysis (1000 repli- trically using the initial renaturation rate method described by cates) were performed using the BioNumerics 5.1 software De Ley et al. (1970). The DNA G+C content of strains KMM package (Applied Maths). A phylogenetic tree was recon- 6277T, KMM 6302, KMM 6303, KMM 6305, KMM 6307, structed using the neighbour-joining method (Saitou & KMM 6308 and KMM 6311 ranged from 35.2 to Nei, 1987), a Jukes & Cantor (1969) correction was 36.1 mol%, which is in the range reported for the recognized applied, and unknown bases were discarded for the anal- species of the genus Lutibacter (Choi & Cho, 2006; Park et al., ysis. The phylogenetic position of our strains was deter- 2010; Lee et al., 2012). The strains under study shared mined from their almost-complete 16S rRNA gene 95–99 % DNA–DNA reassociation with each other, indicat- sequences. The phylogenetic analysis revealed that the ing that the sea urchin isolates belong to a single species novel strains were members of the family Flavobacteriaceae, (Wayne et al., 1987). DNA–DNA relatedness between strain phylum Bacteroidetes. They showed 99.9–100 % 16S rRNA KMM 6277T and its closest relatives, L. litoralis JCM 13034T gene sequence similarity to each other, demonstrating their and L. agarilyticus KCTC 23842T, was 27 and 38 %, respect- afﬁliation to the same species. Strain KMM 6277T was most ively, far below the threshold value of 70 % recommended

1% T Cellulophaga lytica ATCC 23178 (M62796) 100 Salegentibacter salegens DSM 5424T (M92279) 98 Fulvibacter tottoriensis MTT-39T (AB294107) Psychroserpens burtonensis ACAM 188T (U62913) DSW-5T (DQ004686) 100 Polaribacter dokdonensis T 100 Polaribacter filamentus 215 (U73726) 98 Polaribacter irgensii 23-PT (M61002) T 92 Tenacibaculum mesophilum MBIC1140 (AB032501) T 99 Tenacibaculum maritimum NBRC 15946 (AB078057) Maritimimonas rapanae A31T (EU290161) Namhaeicola litoreus DPG-25T (JN033800) 100 T 96 Aestuariicola saemankumensis SMK-142 (EU239499) Actibacter sediminis JC2129T (EF670651) Lutibacter oricola UDC379T (HM031974) Lutibacter holmesii KMM 6277T (JQ241142) 99 99 Lutibacter agarilyticus KYW566T (JN864028) 92 T 86 Lutibacter aestuarii MA-My1 (HM234096) T 81 Lutibacter litoralis CL-TF09 (AY962293) Lutibacter maritimus S7-2T (FJ598048) Lutibacter flavus IMCC 1507T (GU166749) Zobellia galactanivorans DsijT (AF208293)

Fig. 1. Neighbour-joining phylogenetic tree based on 16S rRNA gene sequences showing the phylogenetic position of Luti- bacter holmesii sp. nov. within the genus Lutibacter and family Flavobacteriaceae. It was based on comparison of approximately 1374 nt. Bootstrap values ($70 %) based on 1000 replications are shown at the nodes of the tree. Zobellia galactanivorans DsijT (AF208293) was used as the outgroup. Bar, 0.01 substitutions per nucleotide position.

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Table 1. Cellular fatty acid content of strain KMM 6277T and In order to determine the fatty acid and polar lipid proﬁles, allied species of the genus Lutibacter strain KMM 6277T, L. agarilyticus KCTC 23842T, L. litoralis T T T T JCM 13034 , L. aestuarii CCUG 60022 and L. maritimus Species: 1, KMM 6277 ;2,L. litoralis JCM 13034 ;3,L. aestuarii CCUG 57524T were cultivated on marine agar for 3 days CCUG 60022T;4,L. agarilyticus KCTC 23842T;5,L. maritimus 8 T at 28 C, and the cell mass was harvested during the expo- CCUG 57524 . Data are percentages of total fatty acids. Fatty acids nential growth phase. Fatty acid methyl esters of strain , amounting to 1 % of the total fatty acids in all strains are not KMM 6277T and the reference strains were prepared listed. The strains were grown for 3 days on marine agar 2216. TR, according to the methods described by Sasser (1990) , Traces ( 1 %); –, not detected. All data are from this study. using the standard protocol of the Sherlock Microbial Fatty acid 1 2345 Identiﬁcation System (version 6.0, MIDI) and analysed on a Shimadzu GC-21A chromatograph equipped with a Branched fatty acids fused-silica capillary column (30 m60.25 mm) coated

iso-C14 : 0 1.7 3.9 7.9 8.9 1.8 with Supercowax-10 and SPB-5 phases (Supelco) at iso-C15 : 0 26.2 17.3 23.8 15.5 32.2 210 8C. Cellular fatty acids were identified by using equiv- anteiso-C15 : 0 25.5 11.4 4.1 10.6 5.6 alent chain-length measurements and comparing the reten- iso-C15 : 1 5.1 6.7 6.3 2.4 6.3 tion times with those of authentic standards. The polar T anteiso-C15 : 1 4.0 1.3 1.2 1.0 6.5 lipids of strain KMM 6277 and the reference strains iso-C16 : 0 TR 1.3 – 3.0 1.1 were extracted using the chloroform/methanol extraction iso-C16 : 1 1.3 2.4 TR 3.5 2.9 method of Bligh & Dyer (1959). Two-dimensional TLC v iso-C17 : 1 8c 1.5 – TR TR – of polar lipids was carried out on Merck Kieselgel 60- Straight-chain fatty acids HPTLC (666 cm) using chloroform/methanol/water C15 : 0 5.1 10.6 5.3 5.1 4.2 (65 : 25 : 4, by vol.) in the first dimension and chloro- v C15 : 1 6c 2.6 14.6 3.7 5.3 9.7 form/methanol/acetic acid/water (80 : 12 : 15 : 4, by vol.) C16 : 0 1.4 1.1 1.5 1.6 1.1 in the second dimension (Collins & Shah 1984). The C v6c 1.6 4.1 1.3 1.5 TR 17 : 1 spray reagents used to reveal the spots were 10 % sulfuric C v6c TR TR TR 1.4 – 18 : 1 acid in methanol, molybdate reagent, ninhydrin and a- Hydroxy fatty acids iso-C 3-OH 1.7 6.0 12.1 13.5 4.7 naphthol. Isoprenoid quinones were extracted with chloro- 16 : 0 form/methanol (2 : 1, v/v) and purified by TLC, using a iso-C17 : 0 3-OH 3.0 1.6 5.3 9.4 4.9 mixture of n-hexane and diethyl ether (85 : 15, v/v) as C14 : 0 3-OH TR – 1.2 – – the solvent. Isoprenoid quinone composition was charac- C15 : 0 3-OH 3.4 1.5 3.4 2.5 1.9 C 3-OH TR 1.1 1.0 – – terized by HPLC (Shimadzu LC-10A) using a reversed- 16 : 0 6 Summed feature 3* 12.6 12.2 19.6 12.5 14.2 phase type Supelcosil LC-18 column (15 cm 4.6 mm) and acetonitrile/2-propanol (65 : 35, v/v) as a mobile 2 v 1 *Summed feature 3 consists of iso-C15 : 0 2-OH and/or C16 : 1 7c that phase at a flow rate of 0.5 ml min as described by Koma- could not be separated by the Microbial Identification System. gata & Suzuki (1987). The column was kept at 40 8C. Qui- nones were detected by monitoring at 270 nm and were identified by comparison with known quinones from reference strain Bizionia paragorgiae KMM 6029T. for assigning strains to the same species by Wayne et al. (1987). These results consequently strongly confirmed our proposal The prevalent fatty acids of strain KMM 6277T were iso- that the three strains are genetically different and the novel C15 : 0 (26.2 %), anteiso-C15 : 0 (25.5 %), summed feature isolates represent a novel and distinct species of the genus 3 (comprising iso-C15 : 0 2-OH and/or C16 : 1v7c fatty Lutibacter. acids, 12.6 %), iso-C15 : 1 (5.1 %) and C15 : 0 (5.1 %)

(a)(b)(c)(d)(e)

PE PE 1st PE PE 1st 1st PE 1st 1st L AL L AL AL LLAL AL

2nd 2nd 2nd 2nd 2nd

Fig. 2. Two-dimensional TLC of polar lipids of strain KMM 6277T (a), L. litoralis JCM 13034T (b), L. maritimus CCUG 57524T (c), L. aestuarii CCUG 60022T (d) and L. agarilyticus KCTC 23842T (e). PE, phosphatidylethanolamine; AL, unknown amino lipid; L, unknown lipid.

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Table 2. Differential phenotypic characteristics of Lutibacter Table 2. cont. holmesii sp. nov. and closely related species of the genus Lutibacter Characteristic 1 2 3 4 5 Species: 1, Lutibacter holmesii sp. nov. (n57); 2, L. litoralis JCM + 2 ++ + 13034T;3,L. aestuarii CCUG 60022T;4,L. agarilyticus KCTC Acid phosphatase, 23842T;5,L. maritimus CCUG 57524T. All strains were positive for naphthol-AS-BI- phosphohydrolase catalase, oxidase, alkaline phosphatase and leucine arylamidase activi- b-Glucosidase + 22 ++ ties and hydrolysis of gelatin. All strains were negative for gliding a-Galactosidase 2 + 22 2 motility; arginine dihydrolase, lipase (C14), a-chymotrypsin, b- b-Galactosidase 222 + 2 glucuronidase, a-glucosidase, a-mannosidase and a-fucosidase; pro- N-Acetyl-b- 2 ++ + 2 duction of flexirubin pigments; indole and acetoin production; glucosaminidase hydrolysis of cellulose (CM-cellulose and filter paper) and chitin; DNA G+C content (mol%) 36.1 33.9 30.6 41.6 34.6 acid production from L-arabinose, melibiose, raffinose, L-rhamnose, L-sorbose, trehalose, D-xylose, glycerol, adonitol, inositol, mannitol and sorbitol; utilization of inositol and malonate; susceptibility to gentamicin, kanamycin, neomycin and polymyxin B. Data are from (Table 1). The fatty acid profile of strain KMM 6277T was this study, except data on the DNA G+C content of the recognized consistent with those reported for the recognized species of species of the genus Lutibacter taken from the original descriptions: the genus Lutibacter (Choi et al., 2013; Sung et al., 2015) Choi & Cho (2006), Lee et al. (2012) and Park et al. (2010, 2013). and those of the four reference strains included in this n, Number of strains; A, aerobic; F, facultatively anaerobic; +, posi- study (Table 1). The fatty acid profiles of the type strains of tive; 2, negative; W, weakly positive. all species of the genus Lutibacter demonstrated the presence of iso-C15 : 0 and anteiso-C15 : 0 fatty acids as the dominant Characteristic 1 2 3 4 5 components. However, significant differences in the respect- Type of metabolism A A F A A ive proportions of some fatty acids were found between the Temperature range for 4–31 4–32 4–37 20–28 4–37 novel isolate and its close phylogenetic neighbours (i.e. ante- v growth (8C) iso-C15 : 0,C15 : 1 6c and iso-C16 : 0 3-OH) (Table 1). These Salinity range for 0–4 1–6 0–6 1–4 0.5–5 differences in fatty acid components found between strain T growth (NaCl, %) KMM 6277 and recognized species of the genus Lutibacter Nitrate reduction 222 + 2 support the results of the phylogenetic analysis; therefore, T H2S production W ++ 22 strain KMM 6277 can be considered as representing a sep- Hydrolysis of: arate species within the genus Lutibacter. The polar lipid Aesculin ++2 ++ profile of strain KMM 6277T was composed of phosphatidy- Agar 222 + 2 lethanolamine, one unknown aminolipid and one unknown Casein 22+ 2 + lipid (Fig. 2). The polar lipid composition of the novel strain Starch ++2 + 2 was in line with those of L. aestuarii CCUG 60022T, L. agar- Tween 40 222 2 + ilyticus KCTC 23842T and L. maritimus CCUG 57524T,but Tween 80 222 2 + strain KMM 6277T and the above type strains differed from 2 + 22 2 Urea L. litoralis JCM 13034T by the presence of an unknown lipid. Acid production from: The single respiratory quinone of the novel strain was mena- Cellobiose 2 + 2 ++ 2 ++ + + quinone 6 (MK-6), in line with all other members of the D-Galactose, lactose family Flavobacteriaceae (Bernardet et al.,2002). D-Fructose 2 ++ + + D-Glucose, maltose, 2 ++ + + Phenotypic analysis was performed by using previously D-mannose described methods (Nedashkovskaya et al.,2012).Inaddition, Sucrose 222 2 + API 20NE, API 20E, API 50CH and API ZYM galleries N-Acetylglucosamine 2 ++ + 2 (bioMe´rieux) were used according to the manufacturer’s Utilization of: instructions, except that the gallery was incubated at 28 8C. Cellobiose, D-glucose, 2 ++ + + Susceptibility to antibiotics was examined by the routine D-fructose, mannose diffusion plate method. Discs were impregnated with the 2 ++ + + D-Galactose following antibiotics (mg per disc): ampicillin (10), benzylpe- 2 ++ + + Lactose nicillin (10 U), carbenicillin (100), cefalexin (30), cefazolin + 22 2 Citrate W (30), chloramphenicol (30), erythromycin (15), doxycycline Enzyme activity (10), gentamicin (10), kanamycin (30), lincomycin (15), Esterase (C4) W 2 + 2 W oleandomycin (15), nalidixic acid (30), neomycin (30), oflox- Valine arylamidase W 2 + 2 W acin (5), oxacillin (10), polymyxin B (300 U), rifampicin (5), Esterase lipase (C8) ++22+ streptomycin (30), tetracycline (5) and vancomycin (30). The Cystine arylamidase 22W 2 W Trypsin + 2 + 22 physiological and biochemical characteristics of strain KMM 6277T and other novel strains are listed in the species

Downloaded from www.microbiologyresearch.org by 3994 International Journal of Systematic and Evolutionary Microbiology 65 IP: 157.193.5.232 On: Mon, 01 Aug 2016 09:46:04 Description of Lutibacter holmesii sp. nov. description and in Table 2. The sea-urchin isolates had pheno- English microbiologist, for his huge contribution in the typic characteristics that were consistent with members of the taxonomic study of bacteria belonging to the phylum genus Lutibacter. They were heterotrophic, pigmented, non- Bacteroidetes). motile by gliding and rod-shaped organisms. The sea urchin Cells are heterotrophic, strictly aerobic, Gram-staining- strains possessed oxidase, catalase and alkaline phosphatase negative rods, approximately 0.3–0.8 mm in diameter and activities and hydrolysed gelatin and DNA (Table 2). How- 1.2–5.3 mm in length, non-motile by gliding. On marine ever, sufficient differences between the novel isolates and the agar, colonies are 2–3 mm in diameter, circular, with type strains of species of the genus Lutibacter were observed entire edges, shiny and pale-yellow-pigmented. Growth in acid production from carbohydrates and in utilization of occurs at 4–31 uC (optimum, 23–25 uC) and at pH 5.5– some of them (Table 2). The ability to grow without NaCl, 10.0 (optimum, pH 7.5), and with 0–4 % NaCl (optimum, the absence of urea hydrolysis, acid production from N- 1.5–3 % NaCl). The presence of NaCl is sufficient for acetylglucosamine and a set of enzyme activities can help in growth. Catalase and oxidase activities are present but separating the strains under study from their nearest arginine dihydrolase, lysine decarboxylase, ornithine decar- phylogenetic neighbour, L. litoralis. The absence of caseinase boxylase and tryptophan deaminase activities are absent. and tweenase activities and the presence of amylase Aesculin, gelatin and starch are hydrolysed but agar, casein, activity, citrate utilization and hydrogen sulfide production Tweens 40 and 80, urea, cellulose (CM-cellulose and filter taken together with the absence of oxidation of several paper) and chitin are not. Hydrolysis of DNA and Tween carbohydrates clearly distinguished the isolates from their 20 is strain-dependent. Acid is not produced from L- other close relative, L. maritimus (Table 2). In contrast to L. arabinose, cellobiose, D-fructose, D-galactose, D-glucose, aestuarii, the strains under study could not ferment D-glucose, lactose, maltose, mannose, melibiose, raffinose, L-rhamnose, grow at 37 8C, hydrolyse casein or utilize the majority of sucrose, xylose, N-acetylglucosamine, glycerol or mannitol. carbohydrates (Table 2). The sea-urchin isolates clearly dif- L-Arabinose, glucose, mannose, sucrose, N-acetylglucosamine, fered from their other closest relative, L. agarilyticus, by a set inositol, mannitol and sorbitol are not utilized. Utilization of of physiological and biochemical features including minimal maltose is strain-dependent. In API 20E gallery, weak positive temperature for growth, nitrate reduction, hydrolysis of reaction was observed only for citrate utilization. In the API agar, acid production from cellobiose and N-acetylglucosa- ZYM gallery, esterase (C4), esterase lipase (C8), leucine mine, and several enzyme activities (Table 2). Therefore, on arylamidase, valine arylamidase, trypsin, alkaline phospha- the basis of the data of the comparative analysis presented in tase, acid phosphatase, naphtol-AS-BI-phosphohydrolase this study, the novel strains should be classified in the genus and b-glucosidase activities are present, but lipase (C14), Lutibacter as representing a novel species, for which the cystine arylamidase, a-chymotrypsin, a-andb-galactosidases, name Lutibacter holmesii sp. nov. is proposed. b-glucuronidase, a-glucosidase, N-acetyl-b-glucosaminidase, It is a particularly interesting that the closest phylogenetic a-mannosidase and a-fucosidase activities are absent. Flexir- relatives of the novel strains showing 99.5–99.6 % 16S ubin-type pigments are not formed. Nitrate is not reduced. rRNA gene sequence similarity were identified among the Hydrogen sulphide is produced weakly, but indole and associates of the sea urchins S. intermedius and Strongylocen- acetoin are not. Susceptible to ampicillin, benzylpenicillin, trotus nudus collected from three localities in the northern carbenicillin, chloramphenicol, erythromycin, doxycycline, part of the Sea of Japan (Balakirev et al., 2008). These findings lincomycin and tetracycline; and resistant to gentamicin, taken together with diverse hydrolytic activities of the novel kanamycin, neomycin, oleandomycin, polymyxin B and isolates allow us to suggest that members of the novel species streptomycin. The prevalent fatty acids (w5%) are of the genus Lutibacter, L. holmesii sp. nov., play an important iso-C15 : 0,anteiso-C15 : 0, summed feature 3 (comprising role in metabolic processes of the host-organisms and could iso-C15 : 0 2-OH and/or C16 : 1v7c fatty acids), iso-C15 : 1 and be considered as a part of a resident microflora of the sea C15 : 0. The polar lipid profile is composed of phosphatidy- urchins belonging to the genus Strongylocentrotus. lethanolamine, one unknown aminolipid and one unknown lipid. The major respiratory quinone is MK-6. T T Lutibacter The type strain, KMM 6277 (5 CCUG 62221 5 LMG Emended description of the genus T Choi and Cho 2006 emend. Lee et al. 2012 26737 ), was isolated from the sea urchin Strongylocentrotus intermedius collected from Troitsa Bay, the Sea of Japan. This description is based on those given by Choi & Cho The DNA G+C content of the type strain is 36.1 mol%. (2006) and Lee et al. (2012) except that polar lipids are represented by the major lipid phosphatidylethanolamine, unknown aminolipids and unknown lipids. The DNA Acknowledgements G+C content is 30–42 mol%. We thank Drs Yoshimasa Kosako and Mitsuo Sakamoto (RIKEN Bioresource Center, Ibaraki, Japan) for providing us with the type Description of Lutibacter holmesii sp. nov. strain Lutibacter litoralis JCM 13034T, Professor Jung-Sook Lee (KCTC, Daejeon, Republic of Korea) for the type strain Lutibacter Lutibacter holmesii (hol9me.si.i. N.L. gen. masc. n. holmesii agarilyticus KCTC 23842T, and Professor Edward R. B. Moore of Holmes, named in honour of Barry Holmes, a famous (CCUG, Go¨teborg, Sweden) for the type strains Lutibacter aestuarii

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