International Journal of Systematic and Evolutionary Microbiology (2015), 65, 2172–2178
DOI 10.1099/ijs.0.000238
Streptobacillus felis sp. nov., isolated from a cat
with pneumonia, and emended descriptions of the
genus Streptobacillus and of Streptobacillus
moniliformis
Tobias Eisenberg,1 Stefanie P. Glaeser,2 Werner Nicklas,3
Norman Mauder,4 Matthias Contzen,4 Khayrieh Aledelbi5 and
Peter Kämpfer2
Correspondence
1
Tobias Eisenberg
2
[email protected]
Landesbetrieb Hessisches Landeslabor, D-35392 Giessen, Germany
Institut für Angewandte Mikrobiologie, Justus-Liebig-Universität Giessen, D-35392 Giessen,
Germany
3
Deutsches Krebsforschungszentrum, D-69120 Heidelberg, Germany
4
Chemisches und Veterinäruntersuchungsamt Stuttgart, D-70736 Fellbach, Germany
5
Merlin Diagnostika GmbH, D-53332 Bornheim-Hersel, Germany
A pleomorphic, Gram-stain-negative, rod-shaped, indole-, oxidase- and catalase-negative,
non-spore-forming, non-motile bacterium (strain 131000547T) was isolated from the lungs of a
cat with pneumonia. On the basis of 16S rRNA gene sequence analyses the strain was
assigned to the genus Streptobacillus with 97.6 % sequence similarity to the type strain of
Streptobacillus moniliformis and 94.6 % to that of Streptobacillus hongkongensis. The clear
differentiation of strain 131000547T from Streptobacillus moniliformis and Streptobacillus
hongkongensis was also supported by gyrB, groEL, and recA nucleotide and amino acid
sequence analysis. DNA–DNA hybridization demonstrated #19.9 % (reciprocal 28.7 %)
DNA–DNA relatedness between strain 131000547T and Streptobacillus moniliformis DSM
12112T. Physiological data confirmed the allocation of strain 131000547T to the family
Leptotrichiaceae. Strain 131000547T has a unique profile of enzyme activities allowing
differentiation from the most closely related species. Within the genus Streptobacillus, isolate
131000547T could also unambiguously be separated from Streptobacillus moniliformis and
Streptobacillus hongkongensis by both matrix-assisted laser desorption ionization time-of-flight
mass spectrometry and Fourier transform-infrared spectroscopy. On the basis of these data, a
novel species of the genus Streptobacillus, Streptobacillus felis sp. nov., is proposed with the
type strain 131000547T (5DSM 29248T5CCUG 66203T5CCM 8542T). Emended
descriptions of the genus Streptobacillus and of Streptobacillus moniliformis are also given.
The genus Streptobacillus (Levaditi et al., 1925) (Leptotrichiaceae; Fusobacteriales) comprised, for almost 90 years,
a monotypic species, Streptobacillus moniliformis (Elliott,
Abbreviations: FT-IR, Fourier transform-infrared; MALDI-TOF
MS, matrix-assisted laser desorption ionization time-of-flight
mass spectrometry; RBF, rat bite fever.
The GenBank/EMBL/DDBJ accession numbers for the 16S
rRNA, gyrB, groEL and recA gene sequences of strain
131000547T are HG421076, KP676101, KP657496 and
KP657504, respectively.
Five supplementary figures and four supplementary tables are
available with the online Supplementary Material.
2172
2007; Gaastra et al., 2009), one of the two aetiological
organisms of rat bite fever (RBF), which is an underreported, globally occurring bacterial zoonosis (Gaastra
et al., 2009). The infection is predominantly transmitted
through rat bites and scratches. A second food-borne
form of Streptobacillus moniliformis infection named
Haverhill fever is transmitted by direct or indirect contact
with rat urine (Bleich & Nicklas, 2008; Hayashimoto et al.,
2008; Torres et al., 2003). Acute symptoms of RBF include
fever, malaise, muscle pain, arthritis and abscess formation,
endocarditis, bacteraemia, and maculopapular, petechial or
pustular rash, as well as vomiting and pharyngitis (Gaastra
et al., 2009). Approximately 50–100 % of wild rats
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Streptobacillus felis sp. nov.
asymptomatically carry Streptobacillus moniliformis in their
oropharynx or nasopharynx and shed the organism with
saliva and urine (Ditchfield et al., 1961; Elliott, 2007;
Washburn, 1995), but abscess formation has also been
described in rats and mice (Rohde et al., 2008; Wullenweber
et al., 1990). Apart from rats and mice, other rodent species
such as gerbils, squirrels, spinifex hopping mice or guinea
pigs as well as companion and exotic animals and livestock
are principally susceptible to infection, but mice may
develop clinical disease strain-dependently (Boyer et al.,
1958; Das, 1986; Ditchfield et al., 1961; Gaastra et al., 2009;
Glünder et al., 1982; Gourlay et al., 1982; Mohamed et al.,
1969; Russell & Straube, 1979; Smallwood, 1929; Valverde
et al., 2002; Wullenweber et al., 1990 Yamamoto & Clark,
1966).
In recent years Streptobacillus-like organisms have been
noted, apart from Streptobacillus moniliformis, from which
Streptobacillus hongkongensis (Woo et al., 2014) was recently
described as a novel species, which causes quinsy and septic
arthritis in humans. Two further Streptobacillus species were
reported, one of which was found in a canine oral microbiome project (sequence COT-370) (Dewhirst et al., 2012).
The other is a Streptobacillus species isolated from a cat
with pneumonia (Eisenberg et al., 2014a) and this strain is
the subject of the present description.
Strain 131000547T was originally isolated from a cat lung after
2–5 days of incubation at 37 8C under a capnophilic atmosphere of 10 % (v/v) CO2 on Columbia agar with 5 % sheep
blood (SBA; Oxoid). On this agar, strain 131000547T was
able to grow also at 20–43 8C, but not at 10 or 50 8C. The
strain could also be cultivated on TSA (Tryptone-soy agar,
Oxoid), supplemented with 20 % horse serum, on Schaedler
agar as well as in liquid media [tryptone-soy-bouillon (TSB),
brain heart infusion and peptone broth, supplemented
with 20 % cattle or horse serum], but not on Gassner or MacConkey agar (Oxoid). Good growth could be observed after
48 h. Gram-staining was carried out according to the
Hucker method as described previously (Gerhardt et al.,
1994). Cell morphological features were observed under a
Leitz light microscope (Leitz Diaplan, Leitz; 61000), with
cells grown for 3 days at 37 8C on SBA. Gram staining revealed
irregular Gram-stain-negative, pleomorphic, fusiform to filamentous, non-spore-forming, non-encapsulated, non-acidfast rods, which were arranged in chains and clumps,
sometimes displaying irregular, lateral bulbar swellings.
Single rod-shaped cells were approximately 0.45¡0.1 mm
wide and 0.83¡0.8 mm long.
For phylogenetic analysis genomic DNA was extracted
using a Maxwell 16 FFPE Plus LEV Purification kit
(Promega) according to the manufacturer’s instructions.
The 16S rRNA gene was PCR-amplified as described elsewhere (Edwards et al., 1989). The PCR product was purified
with a QIAquick PCR purification kit (Qiagen) according to
the manufacturer’s instructions and sequenced by GATC
Biotech (Konstanz, Germany). Phylogenetic analysis was
performed in ARB release 5.2 (Ludwig et al., 2004) using
http://ijs.sgmjournals.org
the 16S rRNA-based ‘All-Species Living Tree’ Project
(LTP) database (Yarza et al., 2008) release 108 (July 2012).
All sequences not included in the LTP database were aligned
with the SINA online alignment tool version 1.2.11 (Pruesse
et al., 2012) and implemented in the LTP database. The
alignment of sequences used for tree reconstruction was
controlled manually based on secondary structure information. Pairwise sequence similarities were calculated in
ARB using the ARB neighbour-joining tool without the use
of an evolutionary substitution model. Phylogenetic trees
were reconstructed with the maximum-likelihood method
using RAxML version 7.04 (Stamatakis, 2006) with GTRGAMMA and rapid bootstrap analysis, and with the
maximum-parsimony method using DNAPARS version 3.6
(Felsenstein, 2005). Both trees were based on bootstrap analysis with 100 replications (Felsenstein 1985) and 16S rRNA
gene sequences between sequence termini 103 and 1356
[numbering according to the Escherichia coli rRNA gene
sequence published by Brosius et al. (1978)].
The sequenced 16S rRNA gene fragment of strain
131000547T represents a continuous stretch of 1415 unambiguous nucleotides between sequence positions 9 and
1444 [E. coli numbering (Brosius et al., 1978)].
Strain 131000547T shared highest 16S rRNA gene sequence
similarity with the type strains of Streptobacillus moniliformis (97.6 %) and Streptobacillus hongkongensis (94.5 %),
followed by Sneathia sanguinegens (91.8 %). Sequence
similarities to all other taxa were below 90 %. Independently of the treeing method used, strain 131000547T
formed a distinct cluster (.80 % bootstrap support) with
the type strains of Streptobacillus moniliformis and Streptobacillus hongkongensis (Fig. 1), which was clearly separated
from the genera Sneathia, Sebaldella and Leptotrichia. Strain
131000547T clustered most closely with the type strain of
Streptobacillus moniliformis, which is supported by a high
bootstrap value (100 %). In addition to the type strain of
Streptobacillus moniliformis six further strains of Streptobacillus moniliformis isolated from different original sources
(Table S1, available in the online Supplementary Material)
were analysed in parallel. All shared identical 16S rRNA
gene sequences and did not affect the distinct clustering
of strain 131000547T in the phylogenetic tree.
Employment of the published protocols for Streptobacillus
moniliformis-specific PCR assays, according to Kimura
et al. (2008) and Nicklas (cited in Rohde et al., 2008)
(Table S2) showed that amplification of the specific 16S
rRNA gene sequences for strain 131000547T resulted in
characteristic amplicon sizes of approximately 269 and
1222 bp, respectively.
For further clarification of the phylogenetic relationship of
strain 131000547T to other species of the genus Streptobacillus phylogenetic analyses based on partial nucleotide and
amino acid sequences of gyrB, groEL and recA genes were
performed, according to the analysis described by Woo
et al. (2014). Respective nucleotide sequences were aligned
according to amino acid sequences using CLUSTAL W
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T. Eisenberg and others
0.10
72
*
98 Leptotrichia hongkongensis HKU24T (EU919515)
94 * Leptotrichia trevisanii Wee Tee 1999T (AF206305)
98
Leptotrichia wadei LB 16T (AY029802)
97 * Leptotrichia shahii LB 37T (AY029806)
100
Leptotrichia hofstadii LB 23T (AY029803)
Leptotrichia buccalis DSM 1135T (CP001685)
* Leptotrichia goodfellowii LB 57T (AY029807)
Sebaldella termitidis ATCC 33386T (CP001739)
Streptobacillus moniliformis ATCC 27747 (KP657494)
Streptobacillus moniliformis NCTC 11194 (KP657491)
Streptobacillus moniliformis CIP 55.48T (KP657492)
Streptobacillus moniliformis ATCC 49567 (KP657493)
Streptobacillus moniliformis ATCC 49940 (KP657489)
Streptobacillus moniliformis CIP 81.99T (KP657495)
100 Streptobacillus moniliformis DSM 12112T (CP001779)
86 * Streptobacillus felis sp. nov. 131000547T (HG421076)
* Streptobacillus hongkongensis HKU33T (JQ087389)
100 *
Sneathia sanguinegens CCUG 41628T (AJ344093)
100 * ‘Leptotrichia amnionii’ CCUG 52976 (JQ087392)
Fusobacterium nucleatum subsp. nucleatum ATCC 25586T (AE009951)
Fig. 1. Maximum-likelihood tree showing the phylogenetic position of strain 131000547T within the family Leptotrichiaceae.
The tree was generated in ARB using RAxML (GTR-GAMMA, Rapid Bootstrap analysis, 100 bootstraps) and based on 16S
rRNA gene sequences between positions 103 and 1356 [E. coli numbering; Brosius et al. (1978)]. GenBank accession numbers are given in parentheses. Numbers at branch nodes refer to bootstrap values .70 % (100 replicates). Nodes marked
with asterisks were also present with high bootstrap support (.70 %) in the maximum-parsimony tree. Bar, 0.10 nt substitutions per site.
(Thompson et al., 1994) implemented in MEGA 5 (Tamura
et al., 2011). The correct ORF was obtained by using the
full-length gene sequence of Streptobacillus moniliformis
DSM 12112T as a reference. Pairwise sequence similarities
were calculated based on p-distances (calculated without
an evolutionary model). Phylogenetic trees were generated
using the maximum-likelihood method with a discrete
Gamma-distribution (+G) with five rate categories and by
assuming that a certain fraction of sides are evolutionary
invariable (+I) (for nucleotide sequences) and the Jones–
Thornton–Taylor model (JTT) (Jones et al., 1992)+G+I
(for amino acid sequences). Both trees were based on 100
replications.
In all phylogenetic trees based on partial nucleotide and
more conserved amino acid sequences of gyrB, groEL and
recA the formation of monophyletic clusters was shown,
including all species of the genus Streptobacillus. Strain
131000547T clustered (with high bootstrap support) most
closely, but in a distinct branch, to strains of Streptobacillus
moniliformis (Figs S1–S3). In addition, nucleotide and amino
acid sequence similarities were always considerably lower
between strain 131000547T and strains of Streptobacillus
moniliformis and Streptobacillus hongkongensis (Table S3),
clearly indicating the genetic distinction of strain 131000547T.
For DNA–DNA hybridization, the method described by
Ziemke et al. (1998) was used, except that for nick-translation 2 mg DNA was labelled during 3 h of incubation at
15 8C. The overall DNA–DNA relatedness between strain
131000547T and Streptobacillus moniliformis DSM 12112T
2174
was #19.9 % (reciprocal 28.7 %) as determined by hybridization, and therefore it is evident that they represent separate species.
From the results of sequence analysis of the 16S rRNA, gyrB,
groEL and recA genes and DNA–DNA hybridization, it is
evident that strain 131000547T is different from the genera
Sneathia, Sebaldella and Leptotrichia and from Streptobacillus moniliformis and Streptobacillus hongkongensis.
Results from the physiological characterization are given in
the species description and in Table 1. Extended biochemical
profiling was carried out according to the manufacturer’s
instructions using commercial test systems, i.e. Micronaut
Strep2 especially adapted to the growth characteristics of
Streptobacillus moniliformis (Merlin Diagnostika) for fermentation (Manafi et al., 1991), VITEK2-compact with the
NHI card, and API ZYM (both bioMeriéux). Vitek NHI
identified strain 131000547T, Streptobacillus moniliformis
DSM 12112T and Streptobacillus hongkongensis DSM
26322T as Neisseria cinerea with 99, 98 and 93 % confidence.
Strain 131000547T could be clearly differentiated from Streptobacillus moniliformis DSM 12112T and Streptobacillus
hongkongensis DSM 26322T, although some minor discrepancies with the results of Woo et al. (2014) remain (e.g. for leucine arylamidase and naphthol-AS-BI-phosphohydrolase).
Nevertheless, API ZYM profiles were run with two different
batches, so we believe that the observed differences are
related to the natural variability within Streptobacillus moniliformis or to different batches of API ZYM tests, or are
caused by enzyme differences between strains HKU33T
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Streptobacillus felis sp. nov.
Table 1. Characteristics of strain 131000547T and strains
of
Streptobacillus
moniliformis
and
Streptobacillus
hongkongensis
Taxa: 1, strain 131000547T; 2, Streptobacillus moniliformis DSM 12112T;
3, results from six reference strains of Streptobacillus moniliformis
(ATCC 27747, ATCC 49567, ATCC 49940, NCTC 11194, CIP 55-48
and CIP 81-99); 4, Streptobacillus hongkongensis DSM 26322T. +,
Positive; 2, negative; +/2, variable.
Characteristic
Haemolysis on SBA*
NeuraminidaseD
TripeptidaseD
Proline aminopeptidaseD
Hydroxyproline aminopeptidaseD
Glycyltryptophan aminopeptidaseD
Arginine aminopeptidaseD
PyraseD
Arginine dihydrolaseD
ChitinaseD
Phosphatase (unspecified)d
Phenylalanine arylamidased
Ala-phe-pro-arylamidased
Alkaline phosphatase§
Esterase (C4)§
Esterase lipase (C8)§
Leucine arylamidase§
a-Chymotrypsin§
Acid phosphatase§
Naphthol-AS-BI-phosphohydrolase§
a-Glucosidase§
1
2
3
4
+
2
+
+
+
+
+
+
+/2
+
2
2
2
+
+
+
2
2
+
2
2
2
+
+
+
+
+
+
+
+
2
2
+
+
+
+
+
2
+
+
2
2
+/2
+/2
+
+
+
+
+
+
+
+/2
2
+
+
+/2
+/2
+
+/2
+
+/2
2
2
+
2
2
2
2
2
2
+/2
+
+
+
2
2
+
+
+
2
2
+
+
2
*Data obtained using classical reactions.
DData obtained by Micronaut Strep2 (fermentation) and an individual reaction panel designed for the identification of Streptobacillus
species.
dData obtained using VITEK2-compact with the NHI card.
§Data obtained using the API ZYM system.
and DSM 29248T and DSM 12112T and CCUG13453T. The
antimicrobial susceptibility pattern was determined using
MICs obtained by broth microdilution tests (Merlin Diagnostika). Results were interpreted according to the Clinical
and Laboratory Standards Institute (CLSI; 2012) MIC criteria based on CLSI MIC interpretive standards for other
non-Enterobacteriaceae and anaerobes (2013) according
to Table S4. Strain 131000547T was sensitive to azithromycin, ciprofloxacin, clindamycin, chloramphenicol, erythromycin, gentamicin, meropenem, telithromycin and
tetracycline, but resistant to nalidixic acid and trimethoprim/sulfamethoxazole and had intermediate resistance to
streptomycin. Again, in contrast to the results given by
Woo et al. (2014), Streptobacillus hongkongensis DSM
26322T was a-haemolytic and was the only trimethoprim/
sulfamethoxazole-sensitive strain under study, which
could be confirmed also by E-tests (data not shown).
http://ijs.sgmjournals.org
Apart from the possibilities mentioned above it could be
speculated that this effect could also be attributed to high
levels of p-aminobenzoic acid and thymine/thymidine in
the plating medium, which are known to antagonize sulfamethoxazole and trimethoprim, respectively (Difco
manual).
For matrix-assisted laser desorption ionization time-offlight mass spectrometry (MALDI-TOF MS), strain
131000547T, Streptobacillus moniliformis DSM 12112T, six
reference strains of Streptobacillus moniliformis, Streptobacillus hongkongensis DSM 26322T and Sebaldella termitidis
ATCC 33386T were incubated for 24 h and subsequently
selected from the SBA plates and then subjected to steeltargets, according to the manufacturer’s instructions
(BrukerBiotyper; BrukerDaltonics,). Strains were prepared
using the manufacturer’s direct smear method. Analysis
was performed on a MALDI-TOF MS Biotyper Version
V3.3.1.0. The database used (DB 4613; BrukerDaltonics)
comprised only one entry from Streptobacillus moniliformis
DSM 12112T. Streptobacillus moniliformis DSM 12112T and
all the reference strains of Streptobacillus moniliformis were
identified to the species level with score levels above 2.2.
Strain 131000547T could not be identified correctly, yielding
score levels of between 1.3 and 1.5. Following the manual
inclusion of respective spectra of strain 131000547T to the
database these were most closely related to Streptobacillus
moniliformis. A dendrogram including selected main spectra
peak lists of the family Leptotrichiaceae from the Bruker database, as well as strain 131000547T, Streptobacillus moniliformis DSM 12112T and further reference strains,
Streptobacillus hongkongensis DSM 26322T and Sebaldella
termitidis ATCC 33386T is depicted in Fig. S4.
Fourier transform-infrared (FT-IR) spectroscopy was also
carried out with strain 131000547T, Streptobacillus moniliformis DSM 12112T, Streptobacillus hongkongensis DSM 26322T
and Sebaldella termitidis ATCC 33386T. Bacterial isolates
were cultured independently in five to seven replicates at
37 8C for 48 h on SBA. Harvesting of cells, preparation of bacterial films on zinc selenide plates, drying and handling were
performed as described previously (Kuhm et al., 2009). The
dried bacterial films were used directly for examination by
FT-IR spectroscopy. IR spectra were recorded for each
sample in the transmission mode from 500 to 4000 cm21
with an FT-IR spectrometer (Tensor27 with HTS-XTmodule; BrukerOptics). Acquisition and first analysis of
data were carried out using OPUS software (version 4.2;
BrukerOptics). The IR spectra of tested strains were compared
by cluster analysis [cf. (Eisenberg et al., 2014b; Helm et al.,
1991)]. For cluster analysis, the second derivation of the
vector-normalized spectra in the wave number range of
500–1400 cm21 and 2800–3000 cm21 were used for calculation with Ward’s algorithm (OPUS 4.2) (Ward, 1963).
Comparison of the IR spectra of strain 131000547T with
those from Streptobacillus moniliformis DSM 12112T, Streptobacillus hongkongensis DSM 26322T and Sebaldella termitidis
ATCC 33386T showed a clear separation in two main branches
for the species of the genus Streptobacillus and Sebaldella
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T. Eisenberg and others
termitidis ATCC 33386T. Inside the Streptobacillus branch all
spectra from Streptobacillus moniliformis clustered compactly
together, closely adjacent to strain 131000547T and Streptobacillus hongkongensis DSM 26322T. The dendrogram obtained
depicts the arrangement of isolates in groups according to
their spectral differences (Fig. S5).
Both the molecular differences obtained by DNA–DNA
hybridization and phylogenetic analyses (Fig. 1 and Figs
S1–S3), and the differences based on MALDI-TOF MS
and FT-IR (Figs S4 and S5) support the separate position
of strain 131000547T as a distinct species of the genus
Streptobacillus. The dependence of strain 131000547T to
grow well in a capnophilic environment with 10 % CO2
in the presence of blood or serum, its negative reactivity
for cytochrome oxidase, catalase, nitrate and indole, the
production of a cotton ball-like appearance in liquid
media, its inducible L- forms beside ‘normal’ small butyrous
colonies, its Gram-stain-negative, filamentous, rod-shaped
phenotype arranged in chains and clumps with irregular
bulbar swellings and its broad antimicrobial susceptibilities
(Table 1 and Table S4) also support the placement of the
isolate in the genus Streptobacillus and distinguish it from
Sneathia sanguinegens and ‘Leptotrichia amnionii’ (Woo
et al., 2014). Moreover, genotypic and phenotypic differences strongly support strain 131000547T representing a
novel species, which differs from Streptobacillus moniliformis and Streptobacillus hongkongensis. For these reasons
we propose that strain 131000547T represents a novel
species of the genus Streptobacillus, Streptobacillus felis
sp. nov.
Emended description of the genus
Streptobacillus Levaditi et al. 1925
The description is emended from that given by Staley &
Whitman (2010) and Woo et al. (2014), with the following
features added. Rods with rounded or pointed ends, or
pleomorphic bacilli with coccobacillary, bacillary and filamentous forms. Occur singly or form long, wavy chains.
Gram-stain-negative, non-motile, non-spore-forming. Most
strains are dependent on a capnophilic atmosphere containing 5–10 % CO2 and grow only weakly. Contrarily,
strains from guinea pigs were reported to depend on
anaerobic conditions and one out of two strains of Streptobacillus hongkongensis was able to grow even anaerobically.
Capable of growing on blood agar and weakly on
chocolate agar, but not on MacConkey agar; requires
blood, serum or ascitic fluid for growth. Optimum temperature for growth is 35–37 uC. Most strains are non-haemolytic,
but a Streptobacillus moniliformis strain from a rat with otitis
as well as the type strains of Streptobacillus felis sp. nov. and
Streptobacillus hongkongensis grow with a-haemolysis. Most
strains are positive for esterase (C4) and esterase lipase
(C8). Negative for catalase and cytochrome oxidase, indole
production and nitrate reduction.
The type species is Streptobacillus moniliformis. The DNA
G+C content is 24–26 mol%.
2176
Description of Streptobacillus felis sp. nov.
Streptobacillus felis (fe9lis. L. gen. n. felis of a cat).
Good growth occurs after 2–5 days at 37 uC in a capnophilic
atmosphere of 10 % CO2 on SBA and TSA or in TSB with
20% cattle or horse serum, but only weak growth on Schaedler and chocolate agar and no growth on Gassner and MacConkey agar. In an anaerobic environment reduced growth
is observed. Colonies are tiny, drop-like, shiny and slightly
convex, measuring 0.1–0.4 mm in diameter. Colonies are
a-haemolytic on SBA. Conversion to L -phase or transitional
phase variants may occur spontaneously during cultivation.
In liquid media (e.g. TSB), with the addition of 20 % serum,
growth can be detected after 2–4 days with a typical
‘cotton ball’- or ‘bread crumb’-like appearance. Gramstain-negative, pleomorphic, fusiform to filamentous, nonspore-forming, non-encapsulated, non-acid-fast rods,
0.45+ 0.1 mm (width) and 0.83 + 0.8 mm (length) arranged
in chains and clumps; also sometimes irregular and lateral
bulbar swellings are displayed. Positive for acid phosphatase,
alkaline phosphatase, esterase (C4) and esterase lipase (C8).
Negative for motility, phenylalanine arylamidase, ala-phepro-arylamidase, lipase (C14), leucine arylamidase, valine
arylamidase, cystine arylamidase, trypsin, a-chymotrypsin,
naphthol-AS-BI-phosphohydrolase a-galactosidase, bgalactosidase, a-glucosidase, b-glucosidase, b-glucuronidase,
N-acetyl-b-glucosaminidase, a-mannosidase, a-fucosidase,
cytochrome oxidase, catalase, nitrate and indole.
The type strain, 131000547T (5DSM 29248T5CCUG
66203T5CCM 8542T), was isolated, in Germany, from a
cat with acute suppurative to fibrinous, focally necrotizing
bronchopneumonia with multifocal desquamation of type
II pneumocytes and alveolar macrophages.
Emended description of Streptobacillus
moniliformis Levaditi et al. 1925
The description is emended from that given by Staley & Whitman (2010) and Woo et al. (2014). Rods with rounded or
pointed ends. Occur singly or form long, wavy chains.
Gram-stain-negative, non-motile, non-spore-forming. Conversion to L-phase or transitional phase variants may occur
spontaneously during cultivation. Most strains are dependent
on a capnophilic atmosphere containing 5–10 % CO2 and
grow only weakly anaerobically. Contrarily, strains from
guinea pigs were repeatedly reported to grow exclusively anaerobically. Capable of growing on blood agar and weakly on
chocolate agar, but not on MacConkey agar; requires serum
or ascitic fluid for growth. Optimum temperature for
growth is 35–37 uC. Most strains are non-haemolytic, but a
strain from a rat with otitis grows with a-haemolysis. Ferments glucose to produce acid, but not gas after 3–5 days.
Positive for phenylalanine arylamidase, ala-phe-pro-arylamidase, a-chymotrypsin and esterase lipase (C8). Negative for
naphthol-AS-BI-phosphohydrolase, N-acetyl-b-glucosaminidase, cystine arylamidase, a-fucosidase, a-galactosidase, bgalactosidase, a-glucosidase, b-glucosidase, b-glucuronidase,
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Streptobacillus felis sp. nov.
lipase (C14), a-mannosidase, trypsin, valine arylamidase, catalase, cytochrome oxidase, indole production and nitrate
reduction. Variable for alkaline phosphatase, acid phosphatase, esterase (C4) and leucine arylamidase. Resistant to trimethoprim/sulfamethoxazole (MICw8/152 mg ml21), but
sensitive to azithromycin (50.0625 mg ml21), ciprofloxacin
(51 mg ml21), clindamycin (50.25 mg ml21), chloramphenicol (51 mg ml21), erythromycin (j0.5 mg ml21), gentamicin
(j0.125 mg ml21), meropenem (50.25 mg ml21), nalidixic
acid (52 mg ml21), streptomycin (j1 mg ml21), telithromycin (j0.125 mg ml21) and tetracycline (j0.125 mg ml21).
The type strain is CCUG 13453 (59901 5ATCC 14647
5CCUG 2469T5DSM 12112T5NCTC 10651T). The DNA
G+C content of the type strain is 26.3 mol% (Nolan et al.,
2009).
T
T
T
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Acknowledgements
Helm, D., Labischinski, H., Schallehn, G. & Naumann, D. (1991).
We thank Anna Mohr, Ulrike Kling, Katharina Engel, Andrea ErlesKemna, Bernhard Berkus, Barbara Depner, Anna-Katharina Schmid,
Jana Kistenmacher and Gundula Will for excellent technical assistance
and Barbara Gamb for making even the most exotic manuscripts
available. Hermann Fehrentz is greatly acknowledged for submitting
the cat to necropsy, from which the new species was isolated and
for providing additional rats from his land.
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