International Journal of Systematic and Evolutionary Microbiology (2014), 64, 3660–3667
DOI 10.1099/ijs.0.066274-0
Description of Mariniphaga anaerophila gen. nov.,
sp. nov., a facultatively aerobic marine bacterium
isolated from tidal flat sediment, reclassification of
the Draconibacteriaceae as a later heterotypic
synonym of the Prolixibacteraceae and description
of the family Marinifilaceae fam. nov.
Takao Iino,1 Koji Mori,2 Takashi Itoh,1 Takuji Kudo,1 Ken-ichiro Suzuki2
and Moriya Ohkuma1
Correspondence
Takao Iino
[email protected]
1
Japan Collection of Microorganisms, RIKEN BioResource Center, 3-1-1 Koyadai, Tsukuba,
Ibaraki 305-0074, Japan
2
NITE Biological Resource Center (NBRC), Institute of Technology and Evaluation (NITE),
2-5-8 Kazusakamatari, Kisarazu, Chiba 292-0818, Japan
A mesophilic, chemoheterotrophic bacterium, strain Fu11-5T, was isolated from tidal-flat sediment
from Tokyo Bay, Chiba, Japan. Cells of strain Fu11-5T were facultatively aerobic, Gram-negative,
non-sporulating, non-motile and rod-shaped (1.9–6.9 mm long). Strain Fu11-5T grew optimally at
35–37 6C and pH 6.5–7.0 and with 1–2 % (w/v) NaCl. Oxygen and L-cysteine were used as an
alternative electron acceptor and donor, respectively. Strain Fu11-5T also grew fermentatively on
some pentoses, hexoses and disaccharides and soluble starch. Succinic acid was the major end
product from D-glucose. Phylogenetic analysis based on the 16S rRNA gene sequence revealed
that strain Fu11-5T was affiliated with the order Bacteroidales, and its nearest neighbours were
members of the genera Meniscus, Prolixibacter, Sunxiuqinia, Mangrovibacterium and
Draconibacterium, with 87–91 % sequence similarity. Cell morphology, optimum growth
temperature and utilization of sugars of strain Fu11-5T distinguished the strain from
phylogenetically related bacteria. On the basis of its phenotypic features and phylogenetic
position, a novel genus and species are proposed to accommodate strain Fu11-5T, with the name
Mariniphaga anaerophila gen. nov., sp. nov. The type strain of Mariniphaga anaerophila is strain
Fu11-5T (5JCM 18693T5NBRC 109408T5DSM 26910T). We also propose to combine the
family Draconibacteriaceae into the family Prolixibacteraceae as a later heterotypic synonym and
to place the distinct sublineage of the genus Marinifilum in the family Marinifilaceae fam. nov.
Microbial diversity in nature has been investigated
phylogenetically by both culture-dependent and -independent approaches, and it is obvious that the majority of
micro-organisms have not yet been cultivated in synthetic
laboratory media (Amann et al., 1995; DeLong & Pace,
2001; Olsen et al., 1986; Pace et al., 1986; Ward et al., 1990;
Whitman et al., 1998). In particular, isolation of novel
marine micro-organisms has lagged behind that of
enterobacteria and terrestrial micro-organisms as a result
of their oligotrophic habitats and the difficulty in accessing
sampling sites.
Abbreviations: BI, Bayesian inference; ML, maximum-likelihood; NJ,
neighbour-joining.
The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene
sequence of strain Fu11-5T is AB921558.
3660
In our previous studies, some rare and/or unique microorganisms have been isolated from marine sediment and
the alimentary tracts of marine organisms collected
from the coast and tidal flats in Japan, for example
‘Methanosaeta pelagica’, an acetoclastic and NaCl-requiring
methanogen (Mori et al., 2012), Oscillibacter valericigenes, a
strictly anaerobic, fermentative species that is phylogenetically related to a representative uncultured bacterium
Oscillospira guilliermondii (Iino et al., 2007), Ferrimonas
futtsuensis and Ferrimonas kyonanensis, facultatively anaerobic, selenite-reducing species (Nakagawa et al., 2006), and
others (Hamada et al., 2009, 2010; Muramatsu et al., 2010).
These findings suggest that diverse novel micro-organisms
are still cultivable from the shallow sea and tidal-flat
environments of Japan. Recognition of their diversity and
development of cultivation methods for unknown marine
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Mariniphaga anaerophila gen. nov., sp. nov.
micro-organisms are thus important in understanding
the marine ecosystem and functional diversity in marine
environments.
This paper describes the isolation of a heterotrophic
marine bacterium from sediment of a tidal flat in Tokyo
Bay. On the basis of a polyphasic taxonomic approach, a
novel genus and species are proposed for this bacterium.
Blackening sediment was collected on 10 September 2003
at a depth of approximately 100 cm using a peat sampler
(model DIK-105A; Daiki Rika Kogyo) on the Futtsu tidal
flat in Tokyo Bay, Chiba Prefecture, Japan, as reported
previously (Mori et al., 2012). The soil sample was kept in a
sealed nylon bag with an O2-absorbing and CO2-generating
agent (Anaero-Pack; Mitsubishi Gas Chemical) until it was
inoculated into a fresh medium.
The basal medium used in this study, designated Sw
medium, contained (l21) 0.54 g NH4Cl, 0.14 g KH2PO4,
0.15 g CaCl2 . 2H2O, 4.0 g MgCl2 . 6H2O, 18.0 g NaCl,
2.5 g NaHCO3 and 1.0 ml trace elements solution (Touzel
& Albagnac, 1983), containing 4.0 mg Na2WO4 . H2O and
with NaCl omitted. The medium was adjusted to pH 7.0
with 6 M HCl, and 20 ml medium was dispensed in 50 ml
serum bottles. Dissolved air was removed by flushing with
N2/CO2 (4 : 1, v/v), and the bottles were sealed with butyl
rubber stoppers. Prior to inoculation, 0.2 ml vitamin
solution (Wolin et al., 1963) and 0.2 ml reductant solution
containing 0.5 g Na2S and 0.5 g L-cysteine hydrochloride
l21 were added to each bottle with filtration through
a 0.2 mm-pore membrane filter. For enrichment, 1.0 g
sediment was inoculated into 20 ml HXSw medium
supplemented with 0.2 % (w/v) each yeast extract (Becton
Dickinson) and Polypepton (Nihon Pharmaceutical) in the
Sw medium with filtration and with a headspace of H2/CO2
(4 : 1, v/v; approx. 150 kPa) instead of N2/CO2. The culture
was incubated at 25 uC for 3 weeks and then transferred to
fresh HXSw medium. After several transfers, the culture
was spread on slants of HXSw medium solidified with 1.5 %
(w/v) agar and colonies that appeared were picked and
streaked on fresh HXSw medium. The purification procedure was repeated several times until the culture was deemed
pure, and a culture with uniformly shaped cells was
obtained, which was designated Fu11-5T. After purification,
the isolate was maintained in HXSw medium, and cultivated
routinely in GXSm medium with the addition of 0.2 % (w/v)
yeast extract, 0.2 % (w/v) Polypepton and 10 mM D-glucose
in Sw medium with filtration for the following study.
The isolate was cultivated in 10 ml medium dispensed in
screw-capped tubes (186180 mm, Sanshin Industrial Co.)
with inoculation of a 1/100 dilution of the preculture.
Bacterial growth was followed by the increase in turbidity
at 660 nm determined with a spectrophotometer. Utilization of electron acceptors and donors was determined in
Sw medium without NH4Cl after adding 10 mM pyruvate
and 0.01 % (w/v) yeast extract. Concentrations of electron
acceptors and donors and the product in the cultures were
quantified by HPLC equipped with a conductivity detector
http://ijs.sgmjournals.org
model CDD-10ADsp, a Shim-Pack Cation column IC-C4
and a Shim-Pack Anion column IC-SA2 (Shimadzu).
Utilization of sugars was examined using GXSw medium
with the addition of each sugar in place of D-glucose.
Concentrations of organic acids in cultures were quantified
by HPLC equipped with a diode array detector model SPDM20A (Shimadzu) and Rezex ROA-Organic acid H+ (8 %)
column (Phenomenex).
Cells of strain Fu11-5T were rods, approximately 1.9–6.9 mm
long and 0.5–0.6 mm wide (Fig. 1). Cells usually occurred
singly or in pairs. Motility and spore formation were not
observed under a phase-contrast microscope. Flagellation
was also not observed by electron microscopy. The colony
consistency of strain Fu11-5T was butyrous. Cells stained
Gram-negative by conventional Gram staining.
Strain Fu11-5T was a facultatively aerobic bacterium that
grew better under anaerobic conditions comprising a N2/
CO2 (4 : 1, v/v) atmosphere than under aerobic conditions in
air. Catalase and oxidase reactions were negative. The
growth temperature of strain Fu11-5T was 20–40 uC, with an
optimum at 35–37 uC. No growth was observed at 15 or
45 uC. The pH range for growth was 6.0–8.5, with an
optimum at pH 6.5–7.0. No growth was observed at pH 5.5
or 9.0. Growth occurred in media containing 9 % (w/v)
NaCl or below, with the optimum being 1–2 % (w/v) NaCl.
No growth was observed at 10 % (w/v) NaCl. Strain Fu11-5T
grew using oxygen (5 %, w/v) as the electron acceptor in the
presence of L-cysteine as the electron donor. Sulfate
(10 mM), sulfite (2 mM), thiosulfate (5 mM), elemental
sulfur (1 %, w/v), nitrate (10 mM), nitrite (2 mM) and
iron(III) oxide (2 mM) were not utilized as alternative
electron acceptors in the presence of L-cysteine. Strain Fu115T grew using L-cysteine (2 mM) as the electron donor in
the presence of oxygen (5 %, w/v). H2/CO2 (4 : 1, v/v),
sulfide (2 mM), elemental sulfur (1 %, w/v), thiosulfate
(5 mM), sulfite (2 mM), ammonium (10 mM) and nitrite
(2 mM) were not utilized as alternative electron donors in
the presence of oxygen (5 %, w/v). Strain Fu11-5T grew
(a)
(b)
Fig. 1. Phase-contrast (a) and transmission electron (b) micrographs of cells of strain Fu11-5T. Bars, 5.0 mm (a) and 0.5 mm (b).
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T. Iino and others
fermentatively on L-arabinose, D-ribose, D-xylose, D-glucose,
D-mannose, cellobiose, lactose, maltose, sucrose, trehalose
(all at 10 mM) and soluble starch (0.2 %, w/v). No growth
occurred on D-fructose, D-sorbose, D-mannitol or D-sorbitol
(all at 10 mM). Succinic acid (91.3 %) was the major end
product from D-glucose. Acetic acid (8.7 %) was also
produced as a minor end product. Strain Fu11-5T was
susceptible to ampicillin, chloramphenicol, rifampicin,
tetracycline and vancomycin (all at 100 mg ml21), but
resistant to bacitracin, gentamicin, kanamycin and streptomycin (all at 100 mg ml21).
The major cellular fatty acids of strain Fu11-5T were isoC15 : 0 (26.7 %) and anteiso-C15 : 0 (18.5 %), as determined
by using the MIDI microbial identification system
(Microbial ID; Agilent Technologies) based on the method
described by Sasser (1990). C16 : 0 (7.7 %) was also detected
as a minor component. The major isoprenoid quinone was
identified as menaquinone 7 (MK-7), determined using the
HPLC method described by Komagata & Suzuki (1987).
The polar lipid pattern of strain Fu11-5T comprised only
phosphatidylethanolamine, as determined by using twodimensional TLC with spraying with 5 % ethanolic
molybdophosphoric acid, ninhydrin, Dittmer & Lester
reagent, anisaldehyde reagent and Dragendorff reagent, as
described by Lechevalier et al. (1977) and Minnikin et al.
(1984). The genomic DNA G+C content of strain Fu11-5T
was 41.7 mol%, determined by the HPLC method described by Tamaoka & Komagata (1984).
A 16S rRNA gene of strain Fu11-5T was amplified by PCR
with the universal primers U27F (59-AGAGTTTGATCCTGGCTCAG-39; positions 8–27 in the Escherichia coli
numbering system) and U1492R (59-GGTTACCTTGTTACGACTT-39; positions 1510–1492), and the almostcomplete sequence (1443 bp) was determined using primers
U27F, U1492R, U520F (59-GTGCCAGCAGCCGCGG-39),
U920F (59-AAACTCAAAGGAATTGAC-39), U520R (59-ACCGCGGCTGCTGGC-39) and U920R (59-GTCAATTCCTTTGAGTTT-39). After alignment with the ARB software
(Ludwig et al., 2004), phylogenetic trees were reconstructed
by the neighbour-joining (NJ) method with the CLUSTAL_X
program (Saitou & Nei, 1987; Thompson et al., 1997) and
the maximum-likelihood (ML) method with the MOLPHY
software version 2.3b3 (Felsenstein, 1981; Hasegawa et al.,
1985). In addition, the posterior probabilities of branching
points were estimated by Bayesian inference (BI) using
MrBayes 3.1 (Huelsenbeck & Ronquist, 2001; Ronquist &
Huelsenbeck, 2003), referring to the method and parameters
described by Mori et al. (2008). The topologies of the trees
determined by the three methods were almost identical. In
the phylogenetic trees of the 16S rRNA gene sequences
reconstructed using the NJ, ML and BI methods, strain
Fu11-5T was located near Draconibacterium orientale FH5T
(Du et al., 2014) and formed a distinct sublineage with the
genera Meniscus (Irgens, 1977), Prolixibacter (Holmes et al.,
2007), Sunxiuqinia (Qu et al., 2011), Mangrovibacterium
(Huang et al., 2014) and Draconibacterium (Du et al., 2014)
(Fig. 2). The branching at the base of this group was
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supported by a bootstrap percentage of 99–100 %. The 16S
rRNA gene sequence of strain Fu11-5T showed 87.6–91.3 %
sequence similarity to those of members of the five abovementioned genera.
Morphological, biochemical and physiological properties of
strain Fu11-5T, along with those of members of phylogenetically related genera, are summarized in Table 1. Some
morphological, biochemical and physiological traits of strain
Fu11-5T such as the rod-shaped cells, lack of motility,
facultatively aerobic metabolism, mesophily and neutrophily, were similar to those of the single species in the four
related monospecific genera, namely Meniscus glaucopis
(Irgens, 1977), Prolixibacter bellariivorans (Holmes et al.,
2007), Mangrovibacterium diazotrophicum (Huang et al.,
2014) and D. orientale (Du et al., 2014). However, the
optimum temperature of strain Fu11-5T was slightly higher
than those of these four species. Moreover, strain Fu11-5T
differed from these taxa in the utilization of L-arabinose, Dribose, D-fructose, D-glucose and D-mannose. Strain Fu115T was also different from Sunxiuqinia elliptica (Qu et al.,
2011) and Sunxiuqinia faeciviva (Takai et al., 2013) in
morphological and physiological properties, e.g. cell morphology, optimum growth temperature and utilization of
sugars, as shown in Table 1.
On the basis of its distinct phylogenetic position, morphology and biochemical and physiological properties, strain
Fu11-5T was distinguished from members of the five related
genera Draconibacterium, Mangrovibacterium, Meniscus,
Prolixibacter and Sunxiuqinia. Consequently, a novel species
in a new genus, Mariniphaga anaerophila gen. nov., sp. nov.,
is proposed to accommodate strain Fu11-5T.
Recently, the two families Prolixibacteraceae and Draconibacteriaceae have been proposed for two sublineages: the
former accommodates Prolixibacter, Mangrovibacterium
and Sunxiuqinia (Huang et al., 2014) and the latter
accommodates Draconibacterium (Du et al., 2014). In the
phylogenetic tree reconstructed by Du et al. (2014), the
genus Draconibacterium was indicated to form a distinct
sublineage that branched at the periphery of the five
known families in the order Bacteroidales with the
genera Mangrovibacterium, Marinifilum, Prolixibacter and
Sunxiuqinia by a bootstrap percentage of 91 %, but the
phylogenetic independence of the genus Draconibacterium
from the genus Marinifilum and other related genera in this
sublineage was not well supported, as shown by the low
bootstrap percentage of 63 %. This implies that the family
Draconibacteriaceae should not be retained for the single
genus Draconibacterium and the species D. orientale. In our
study, 16S rRNA gene sequence analysis revealed that the
genera Meniscus (Irgens, 1977), Prolixibacter (Holmes et al.,
2007), Sunxiuqinia (Qu et al., 2011), Mangrovibacterium
(Huang et al., 2014), Draconibacterium (Du et al., 2014)
and Mariniphaga gen. nov. formed a distinct phylogenetic lineage adjacent to the families Bacteroidaceae,
Porphyromonadaceae, Prevotellaceae, Marinilabiliaceae and
Rikenellaceae and the genus Marinifilum in the order
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Mariniphaga anaerophila gen. nov., sp. nov.
Family Bacteroidaceae
0.02
Family Prevotellaceae
Family Porphyromonadaceae
B
A
C
Sunxiuqinia elliptica DQHS-4T (GQ200190)
Sunxiuqinia faeciviva JAM-BA0302T (AB362263)
Mangrovibacterium diazotrophicum SCSIO N0430T (JX983191)
Meniscus glaucopis ATCC 29398T (GU269545)
Prolixibacter bellariavorans F2T (AY918928)
Mariniphaga anaerophila Fu11-5T (AB921558)
Draconibacterium orientale FH5T (JQ683778)
Marinifilum fragile JC2469T (FJ394546)
Marinifilum flexuosum CECT 7448T (HE613737)
Lineage 1
Lineage 2
Family Marinilabiliaceae
Family Rikenellaceae
Class Flavobacteriia
Class Sphingobacteriia
Class Cytophagiia
Chlorobium limicola DSM 245T (CP001097)
Fig. 2. Phylogenetic tree of strain Fu11-5T and strains of related species based on 16S rRNA gene sequences. The tree was
based on an alignment of 1296 bp of the 16S rRNA gene sequence and reconstructed by using the NJ method. Open circles at
branching nodes indicate supporting probabilities above 95 % by all the phylogenetic analysis methods (NJ, ML and BI), and
solid circles indicate probabilities above 85 % by two or more analyses. Bootstrap probabilities at nodes A, B and C are as
follows (NJ/ML/BI): A, 68/”/”; B, 99/99/100; C, 100/100/100. Lineage 1 represents the Prolixibacteraceae as defined in this
study, and lineage 2 represents the Marinifilaceae fam. nov. Bar, 0.02 substitutions per nucleotide position.
Bacteroidales (Fig. 2). This lineage was strongly supported
with high bootstrap values by the three phylogenetic analysis
methods (99–100 %). The cell morphology, oxygenic
metabolism, major respiratory quinones and DNA G+C
contents of these six genera are similar (Table 2). Therefore,
the two lineages named Prolixibacteraceae and Draconibacteriaceae should be combined, and the name Prolixibacteraceae should be retained according to Rule 38 of the
Bacteriological Code to avoid taxonomic complication and
to keep priority of publication (Lapage et al., 1992).
Fundamentally, the genus Meniscus (Irgens, 1977) should
be assigned as the type genus of the taxon named
Prolixibacteraceae rather than Prolixibacter (Holmes et al.,
2007) according to Principle 8 of the Bacteriological Code.
http://ijs.sgmjournals.org
However, the name Prolixibacteraceae is retained because
this name is not illegitimate since the preceding proposal
was based on a phylogenetic analysis without Meniscus
glaucopis (Huang et al., 2014). Consequently, we propose to
combine the family Draconibacteriaceae into the family
Prolixibacteraceae. The name Draconibacteriaceae Du et al.
2014 would be regarded as a later heterotypic synonym of
Prolixibacteraceae Huang et al. 2014. On the other hand, the
genus Marinifilum was completely separated from all known
families in the order Bacteroidales in the phylogenetic trees
reconstructed by three methods (Fig. 2). We therefore
propose a novel family for the distinct lineage accommodating the genus Marinifilum, to be named Marinifilaceae
fam. nov.
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T. Iino and others
Table 1. Morphological, biochemical and physiological properties of strain Fu11-5T and phylogenetic relatives
Taxa: 1, Mariniphaga anaerophila gen. nov., sp. nov. Fu11-5T (data from the present study); 2, Draconibacterium [one species (n51); Du et al.,
2014]; 3, Mangrovibacterium (n51; Huang et al., 2014); 4, Meniscus (n51; Irgens, 1977); 5, Prolixibacter (n51; Holmes et al., 2007); 6, Sunxiuqinia
(n52; Qu et al., 2011; Takai et al., 2013). +, Positive; V, variable; 2, negative; ND, no data available.
Characteristic
Cell form
Cell length (mm)
Growth optima
Temperature (uC)
pH
NaCl (%, w/v)*
Substrates for growth
L-Arabinose
D-Ribose
D-Xylose
D-Fructose
D-Glucose
D-Mannose
Lactose
Maltose
Sucrose
Trehalose
D-Mannitol
D-Sorbitol
Soluble starch
DNA G+C content (mol%)
1
2
3
4
5
6
Rods
2–7
Rods
1–2
Rods
5–6
Rods
2–3
Rods
10–12
Elliptical and rods
0.8–28
35–37
6.5–7.0
1–2
28–32
7.0–7.5
2–4
25–30
7.0
0.5
30
7.0
22
7.0
2
30
7–8
2–3
+
+
ND
ND
+
+
+
+
+
2
+
+
+
+
2
2
2
2
+
2
+
+
+
+
+
+
2
2
+
44.9
2
+
+
+
+
2
+
+
+
+
+
+
2
2
+
41.7
+
+
+
ND
ND
ND
ND
2
+
ND
ND
+
ND
+
+
ND
ND
ND
2
ND
ND
+
42.0
ND
ND
43
44.9
V
V
2
2
2
2
2
+
+
+
2
43.5
*Values indicate NaCl concentrations added to the basal medium.
form rods. Catalase-negative and oxidase-negative. The
major cellular fatty acids are iso-C15 : 0 and anteiso-C15 : 0.
The major respiratory quinone is MK-7. The major polar
lipid is phosphatidylethanolamine. The genomic DNA G+C
content of the type strain of the type species is 41.7 mol%.
The genus represents a distinct phylogenetic lineage in the
family Prolixibacteraceae based on 16S rRNA gene sequence
analysis. The type species is Mariniphaga anaerophila.
Description of Mariniphaga gen. nov.
Mariniphaga (Ma.ri.ni.pha9ga. L. adj. marinus marine; Gr.
v. phagein to devour, to eat; N.L. fem. n. Mariniphaga a
marine eater).
Facultatively aerobic, mesophilic, neutrophilic, moderately
halophilic and chemo-organoheterotrophic bacteria. Gramstaining negative, non-sporulating and non-motile. Cells
Table 2. Taxonomic properties of the seven families of the order Bacteroidales
Families: 1, Bacteroidaceae [six genera (n56); Krieg et al., 2011]; 2, Marinifilaceae fam. nov. (n51; Na et al., 2009; Ruvira et al., 2013); 3,
Marinilabiliaceae (n510; Gao et al., 2013; Krieg et al., 2011; Miyazaki et al., 2012; Sorokin et al., 2011; Yang et al., 2014; Zhao & Chen, 2012; Zhao
et al., 2012); 4, Porphyromonadaceae (n512; Hardham et al., 2008; Jabari et al., 2012; Krieg et al., 2011; Sakamoto & Benno, 2006; Sakamoto et al.,
2009; Shkoporov et al., 2013); 5, Prevotellaceae (n54; Downes et al., 2013; Krieg et al., 2011; Moore & Moore, 1994; Morotomi et al., 2009);
6, Prolixibacteraceae (n56; Du et al., 2014; Holmes et al., 2007; Huang et al., 2014; Irgens, 1977; Qu et al., 2011; Takai et al., 2013; present study);
7, Rikenellaceae (n53; Abe et al., 2012; Krieg et al., 2011). Members of all families stain Gram-negative and are non-spore-forming.
Characteristic
Cell form
Oxygenic metabolism*
Major respiratory menaquinone(s)
DNA G+C content (mol%)
*A, Aerobic;
3664
AN,
anaerobic;
FAN,
1
2
3
4
5
Rods
Filamentous
AN
FAN
10, 11
34–62
7
33–45
7
37–44
6
7
Rods
Rods
FAN–AN
FAN–AN
Rods
Rods
Rods
AN
A–FAN
8, 9, 10, 11, 12
37–55
AN
8, 9, 10, 11, 12, 13
40–58
7
42–45
8
55–61
facultatively anaerobic.
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Mariniphaga anaerophila gen. nov., sp. nov.
Description of Mariniphaga anaerophila sp. nov.
Acknowledgements
Mariniphaga anaerophila (an.ae.ro9phi.la. Gr pref. an non-,
not; Gr. n. aer, aeris air; N.L. fem. adj. phila from Gr. fem.
adj. philê friend to, loving; N.L. fem. adj. anaerophila not
air-loving).
The authors thank Mr Takahiro Iwami (NITE) for technical support.
We also thank Dr Mitsuo Sakamoto (RIKEN-BRC JCM) for helpful
discussions on several aspects of the paper. This work was supported
by a Grant-in-Aid for Young Scientists from the Japan Society for the
Promotion of Science (no. 23770095).
The following properties are given in addition to the genus
description. Cells are 0.5–0.661.9–6.9 mm. Growth occurs
at 20–40 uC with an optimum at 35–37 uC. The pH range
for growth is 6.0–8.5 with an optimum around pH 6.5–7.0.
Growth occurs below 9 % (w/v) NaCl, with an optimum at
1–2 % (w/v) NaCl. Oxygen [5 % (v/v) in N2] is used as an
alternative electron acceptor. Sulfate, sulfite, thiosulfate,
elemental sulfur, nitrate, nitrite and iron(III) oxide are not
used as alternative electron acceptors. L-Cysteine is used as
an alternative electron donor. H2/CO2 (4 : 1, v/v), sulfide,
elemental sulfur, thiosulfate, sulfite, ammonium and nitrite
are not used as alternative electron donors. Fermentative
growth occurs on L-arabinose, D-ribose, D-xylose, Dglucose, D-mannose, cellobiose, lactose, maltose, sucrose,
trehalose and soluble starch. No growth occurs on Dfructose, D-sorbose, D-mannitol or D-sorbitol.
The type strain is Fu11-5T (5JCM 18693T5NBRC
109408T5DSM 26910T), which was isolated from sediment
from a tidal flat in Futtsu, Chiba, Japan.
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Marinifilaceae (Ma.ri.ni.fi.la9ce.ae. N.L. neut. n. Marinifilum
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gene sequence. The family accommodates the genus Marinifilum. Gram-staining-negative, filamentous, non-sporulating, non-motile, facultatively anaerobic and chemoorganoheterotrophic. The major respiratory quinone is
MK-7. The type genus is Marinifilum.
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