Content uploaded by Klaus P. Schaal
Author content
All content in this area was uploaded by Klaus P. Schaal
Content may be subject to copyright.
~NTERNATIONAL
JOURNAL
OF
SYSTEMATIC
BACTERIOLOGY,
Apr. 1993,
p.
266-271
(Copyright
0
1993, International Union
of
Microbiological Societies
O020-7713/93/020266-06$02.00/0
Vol.
43,
No.
2
A
New
Actinomycete Species,
Nocardiopsis lucentensis
sp.
nov.
A.
F.
YASSIN,l*
E.
A.
GALINSKI,2
A.
WOHLFARTH,, K.-D. JAHNKE,3
K.
P. SCHAAL,l
AND
H.
G. TRUPER,
Institut
fur
Medizinische Mikrobiologie und Immunologie der Universitat Bonn, Sigmund-Freud Strasse
25,
and Institut
fur
Mikrobiologie und Biotechnologie der Universitat Bonn,
5300
Bonn
1,
and Deutsche
Sammlung von Mikroolganismen und Zellkulturen GmbH,
3300
Braunschweig, Germany
A
new species of the genus
Nocardiopsis,
for which we propose the name
Nocardiopsis
lucentensis sp. nov.
(type strain, strain DSM
44048),
was isolated from a salt marsh soil sample near Alicante, Spain. Whole-cell
hydrolysates contain the
meso
isomer of diaminopimelic acid and no characteristic sugar; thus, the cell wall
composition is type 111. Menaquinone
MK-lO(H,)
is the major menaquinone, and the phospholipid type is type
PI11 (phosphatidylcholine present). Spore chains are rectiflexibilis, and in the early stages
of
sporulation
zig-zag-shaped aerial hyphae are observed. This microorganism produces compatible solutes of the ectoine type
and is characterized by a yellowish to yellowish brown substrate mycelium and a white aerial mycelium. This
organism exhibits only
40
to
50%
DNA relatedness to other
Nocardiopsis
spp.
In 1904 Brocq-Rousseau (4) created the name
Streptothrix
ldassonvillei
to designate a strain isolated from mildewed
grain. Some years later, Liegard and Landrieu (33) isolated a
microorganism from a case of conjunctivitis which they
considered to be identical to
Streptothrix dassonvillei
but
which they assigned to the genus
Nocardia.
Gordon and
Horan (18) discovered that the macroscopic appearance and
a number of the physiological characteristics
of
Nocardia
dassonvillei
were similar to those of
Streptomyces griseus.
Subsequently, Lechevalier and Lechevalier
(30)
described
the genus
Actinomadura
to harbor
Nocardia madurae
(Vin-
cent) Blanchard (the type species),
Nocardia pelletieri
(La-
veran) Pinoy, and
Nocardia dassonvillei
(Brocq-Rousseau)
Liegard and Landrieu. However, striking morphological
differences of
Actinomadura dassonvillei
(e.g., the mode of
sporulation of the aerial mycelium, the nature
of
the sub-
strate mycelium, and the lack of madurose in whole-cell
hydrolysates) led Meyer to transfer this species to a new
genus,
Nocardiopsis,
as
Nocardiopsis dassonvillei
(35). The
genus
Nocardiopsis
was described to comprise strains that
exhibit fragmenting substrate and aerial mycelia and have
cell wall chemotype I11
(meso
isomer
of
diaminopimelic acid
and no characteristic sugar in whole-cell hydrolysates).
An
earlier description of the genus included microorganisms
with a type PI11 phospholipid pattern sensu Lechevalier et
al. (32) (phosphatidylcholine as a diagnostic phospholipid),
as well as microorganisms with a type PIV phospholipid
pattern (glucosamine-containing phospholipid). In recent
studies (20, 21, 24, 28), strains which possess type PIV
phospholipids, including
Nocardiopsis coeruleofusca
(41,
42),
Nocardiopsis flava
(13,41,42),
Nocardiopsis longispora
(41, 421,
Nocardiopsis mutabilis
(43),
Nocardiopsis mutabi-
lis
subsp.
cryophilus
(49, and
Nocardiopsis yringae
(14),
were found to contain rhamnose and large amounts of
galactose in whole-cell hydrolysates (the sugar pattern char-
acteristic of the genus
Saccharothrix
[29]) and therefore
were reclassified in the genus
Saccharothrix.
Another spe-
cies,
Nocardiupsis afncana,
was found to belong to the
genus
Microtetraspora
(25).
Phenetic data indicated that the
single remaining species in the genus,
Nocardiopsis antarc-
tica
(l),
is
identical to the type strain of
Nocardiopsis
dassonvillei
(36).
To
maintain the integrity of the genus
Nocardiopsis,
only those actinomycete isolates should be
*
Corresponding author.
included in the genus which possess meso-diaminopimelic
acid as the cell wall diamino acid, have no characteristic
sugar in whole-cell hydrolysates and no mycolic acids in
whole-cell methanolysates (cell wall chemotype 111), have
phosphatidylcholine as a characteristic phospholipid (phos-
pholipid type 111), have fatty acid profiles which comprise
saturated, unsaturated, iso, anteiso, and tuberculostearic
acids, have menaquinones with
10
isoprene units with a high
degree of hydrogenation as major menaquinones, and have
DNAs with G+C contents of 60 to 70 mol%.
In the course of isolation of microorganisms from uncom-
mon ecological niches, some unusual filamentous bacteria
were isolated from salt marsh soils collected near Alicante,
Spain. These organisms had chemotaxonomic and morpho-
logical characteristics which justify their placement in the
genus
Nocardiopsis.
However, the physiological behavior of
these microorganisms on different media, their inability
to
grow in basal International
Streptomyces
Project (ISP) me-
dium 9, but only when it was supplemented with
5
to 10%
NaCl, and their formation
of
aerial hyphae on different ISP
media only when they were supplemented with
5
to 10%
NaCl led
us
to propose the name
Nocardiopsis lucentensis
sp. nov. for strain
MATERIALS AND METHODS
Bacterial strains and culture conditions.
The strains used in
this study include
Nocardiopsis dassonvillei
DSM 43111T
(T
=
type strain) (2, 4, 30,
33,
39,
Nociardiopsis alborubida
DSM 40465T
(12,
21),
Nocardiopsis listen
DSM 40297T
(10,
21,
48),
Nocardiopsis alba
subsp.
alba
DSM 43119 (2, 21,
35), and
Nocardiopsis alba
subsp.
prasina
DSM 43845T (21,
40). Strains
A5-1T
and A4-* were isolated from a soil sample
collected in a salt marsh area near Alicante, Spain. The
strains were obtained from plates of mineral salts-starch
medium containing the following ingredients:
0.5
g of
of CaCl,
.
2H20, 0.01 g
of
FeSO,
.
7H20, 100 g
of
NaC1,
10.0 g of starch, and 1,000.0 ml of distilled water (pH 7.5).
The plates were incubated at 30°C for 7 days, and the isolates
were then subcultured on the same medium. Stock cultures
were grown on inorganic salts-starch agar (ISP medium 4).
The other media used in this study were those recommended
for use by the ISP (44) and include yeast extract-malt extract
agar (ISP medium 2), oatmeal agar (ISP medium 3), and
glycerol-asparagine agar (ISP medium
5).
K2HPO4,O.l g
of
MgSO4
.
7H,O,
3.0
g
of
NH4HCO,,O.O5 g
266
VOL.
43, 1993
NEW ACTINOMYCETE SPECIES
267
Morphology and pigmentation.
The organism was grown
on ISP medium 2, ISP medium 3, ISP medium 4, and
ISP
medium 5, as described by Shirling and Gottlieb (44), and
was examined for pigmentation, aerial mycelium, and other
morphological features. Cultures were grown for 4 weeks,
and observations were made at weekly intervals. Slide
preparations from 4-, 7-, or 14-day-old cultures were exam-
ined for the presence of spore chains. Air-dried smears from
ISP medium 4 were stained by Gram's method and a
modification of the Ziehl-Neelsen method in order to deter-
mine the Gram reaction and acid fastness, respectively. The
micromorphology was determined with a culture grown at
37°C for
10
days on ISP medium 4. Preparations of whole
cells were stained with fuchsin and safranin and examined
under a light microscope at a magnification of ~700. Elec-
tron micrographs were made with a Leitz model MMR
1600
scanning electron microscope. Samples of 14-day-old colo-
nial growth on
ISP
medium 4 supplemented with 10% NaCl
were prepared by cutting agar blocks from the growth
medium, fixing them with glutaraldehyde, and dehydrating
them by using a graded methoxyethanol series and finally
100% acetone. The dehydrated sections were critical point
dried, mounted on aluminum stubs, and then sputter coated
with gold-palladium.
Physiological tests.
The physiological tests used to charac-
terize strains A5-lT and A4-2 were those of Gordon (15, 16)
and Gordon and Mihm
(19).
The tests used to determine
carbohydrate utilization and the test used to determine
melanoid pigments were those of Shirling and Gottlieb (44).
Catalase production was tested by mixing
1
loopful
of
a
1-week-old culture in glucose-nutrient agar with
1
drop of
freshly prepared
5%
hydrogen peroxide. Phosphatase activ-
ity was determined by the methods of Kurup and Schmitt
(27). The release of o-nitrophenol from o-nitrophenyl-P-D-
galactopyranoside and
p-nitrophenyl-P-D-glucoside
was
used to assay P-galactosidase and P-glucosidase activities,
respectively, by the method
of
Tsukamura (46). Degradation
of
elastin was determined by the disappearance of the
substrate (0.3%, wt/vol) around the area
of
growth by using
nutrient agar as a basal medium; the preparations were
incubated for 10 days. Arbutin degradation was determined
by the method of Kurup and Schmitt (27). To determine
lysozyme susceptibility, a
0.05%
(wt/vol) solution
of
ly-
sozyme was sterilized by membrane filtration and added to
autoclaved Czapek Dox-yeast extract-Casamino Acid broth
(7) to give a final concentration of 0.0025% (wt/vol). Small
tubes
of
medium with and without lysozyme were inoculated
with
1
loopful
of
an old culture and examined for growth for
up to 10 days. Strains which grew in both the control and test
media were scored as positive for lysozyme resistance (17).
Tolerance to salt was determined by growing the organisms
on glucose-nutrient agar plates supplemented with
5
and
10%
NaCl. The susceptibilities
of
the organisms to various anti-
biotics were studied on nutrient agar plates containing vari-
ous
concentrations of the drugs. The drugs were aseptically
mixed with sterile molten agar, and the preparations were
maintained at 45°C and poured into plates. After inoculation,
the plates were incubated at 37°C for
1
week. Growth on the
media was compared with growth of a control and was
recorded as negative or positive (no growth or growth,
respectively). Various concentrations
(0.03
to 128 p,g/ml) of
penicillin G, neomycin, lincomycin, gentamicin, rifampin,
and streptomycin were tested.
Cell chemistry.
Analysis
of
whole-cell hydrolysates for
characterization of amino acids and sugars was performed
by the methods of Becker et al. (3) and Lechevalier (31).
Cellular fatty acid and mycolic acid methyl esters were
prepared from whole-cell methanolysates by the method of
Minnikin et al. (37, 38); the presence
of
both fatty acid and
mycolic acid methyl esters was detected by thin-layer chro-
matography, and the fatty acid methyl ester profile was
determined by gas-liquid chromatography. Menaquinones
were extracted and purified by the methods
of
Collins et al.
(6),
and menaquinone composition was determined by using
a Finnigan Mat 212 mass spectrometer. Phospholipids were
extracted by the method of Minnikin et al.
(39)
and were
determined by two-dimensional thin-layer chromatography,
using chloroform-methanol-water
(65
:
25
:
4, vol/vol) in the
first dimension followed by chloroform-acetic acid-metha-
nol-water (40: 7.5
:
6
:
2, vol/vol) in the second dimension. The
phospholipids were visualized by spraying the preparation
with the phosphate stain
of
Dittmer and Lester
(9)
and were
tentatively identified by using phospholipid standards. Phos-
phatidylcholine was identified with Dragendorf's reagent
DNA isolation and characterization.
DNA was isolated by
chromatography on hydroxyapatite by using the method of
Cashion et al. (5). The G+C contents were determined by
high-performance liquid chromatography (34). DNA-DNA
hybridizations were carried out by using the method
of
De
Ley et al.
(8)
as modified by Huss et al. (22). The concen-
tration of DNA was
45
pg/ml. Because
of
the high G+C
content of the DNAs, 20% (vol/vol) dimethyl sulfoxide (11)
was added to 2.5~ standard saline citrate (Ix standard
saline citrate is 0.15 M NaCl plus 0.015 M sodium citrate) to
depress the melting point
(T,)
of the DNA. The resulting
buffer contained
2~
standard saline citrate and 20% dimethyl
sulfoxide. The average
T,
was determined from the DNA
melting curves, and the optimal temperature of renaturation
(TOR) was calculated as follows: TOR
=
T,,,
-
25°C. Levels
of DNA-DNA relatedness were determined from the rena-
turation rates, which were calculated by regression analysis
of data obtained between 10 and 30 min after the start
of
the
reaction, using the computer program TRANSFER.BAS
(23). The DNA relatedness values
(H)
were calculated by the
following formula:
H
=
[(4
x
VM)
-
VA
-
VB]/(2
X
dVA
x
VB), where V is the renaturation rate, A is DNA,, B
is
DNA,, and M is a
1:l
mixture of DNA, and DNA,.
(47)-
RESULTS AND DISCUSSION
Strain A,-,= has microbiological properties consistent with
its assignment to the genus
Nocardiopsis
(35,
36). It is an
aerobic, gram-positive, non-acid-fast, nonmotile organism
which forms an extremely branched substrate mycelium that
fragments into rod and coccal elements. The substrate
mycelium bears the aerial hyphae, which frequently form
chains of smooth spores (Fig. 1). During spore formation
hyphae have a zig-zag appearance because
of
developing
spore subtending angles
of
various degrees with respect to
the neighboring spores.
Strain A5_1T has chemotaxonomic characteristics which
support its placement in the genus
Nocardiopsis.
The cell
wall contains meso-diaminopimelic acid as the diamino acid
and possesses no characteristic sugars (wall chemotype 111);
the organism lacks mycolic acids, contains major amounts
of
is0
and anteiso branched-chain fatty acids, as well as sub-
stantial amounts
of
tuberculostearic acid, has menaquinones
with 10 isoprene units, with the octahydrogenated form
MK-lO(H,) as the major menaquinone, and has phospholipid
type PI11 sensu Lechevalier et al. (32) (phosphatidylcholine
as the characteristic phospholipid).
268
YASSIN
ET
AL.
INT.
J.
SYST.
BACTERIOL.
FIG.
1.
Scanning electron micrograph
of
strain
A5-lT,
showing zig-zag hyphae and spore chain with a smooth surface.
The
culture was
grown on
ISP
medium 4 supplemented with 10%
NaCl
for 14 days at
35°C.
Bar
=
1.0
p,m.
Identity
of
species.
A total of five strains of
Nocardiopsis
spp. were grown along with strain A5-iT on different media to
study
56
physiological characteristics (Tables
1
and
2).
Although these strains share many characteristics, they can
be separated into two groups on the basis of the ability to
grow in ISP medium 9 and utilize different carbon sources.
The first group includes strain A5-lT,
Nocardiopsis alboru-
bida
DSM 40465T,
Nocardiopsis listen
DSM 40297T, and
Nocardiopsis alba
subsp.
prasina
DSM 43845T, which were
not able to grow in ISP medium 9 and to utilize carbon
sources unless the medium was supplemented with
5%
NaCl
(Table
2).
The second group includes
Nocardiopsis dasson-
villei
DSM 43111T and
Nocardiopsis alba
subsp.
alba
DSM
43119, which were able to grow and to utilize carbon sources
in the presence or absence of NaCl (Table 2). Tables
1
and 2
list the differences between strain A5-lT and other members
of the two groups.
The G+C content
of
the DNA of strain A,-,T was 71.0
mol%. Table
3
shows the levels of DNA relatedness between
strain A5-1T and the other strains, as determined by the
optical method. The DNA of strain exhibits levels of
relatedness ranging from 40 to
50%
with the DNAs
of
other
Nocardiopsk
species. These values indicate the homogene-
ity
of the genus
Nocardiopsis.
This is in contrast to the
previous description of Meyer (36), which was based on an
old description of the genus.
Together, these differences suggest that strain
As-lT
is
unlikely to be a previously described
Nocardiopsis
species.
Thus, we propose for it the name
Nocardiopsis lucentensis.
A description is given below.
Nocardiopsis lucentensis
sp.
nov.
Nocardiopsis lucentensis
(lu. cen. ten’sis.
M.
L.
adj,
lucentensis,
referring to Lucen-
tum, the ancient Latin name of Alicante, a city in Spain,
where the type strain was isolated). Aerobic, gram-positive,
non-acid-fast, nonmotile, filamentous actinomycete.
Cultural characteristics.
Cultures of strain A5.1T grow well
on both complex and defined media. Aerial hyphae are
initiated when NaCl at
a
concentration of up to 10% is added
to the growth medium. The aerial spore mass color is
predominantly white. The reverse side of culture growth
is
yellow to yellowish brown. No distinctive pigmentation
is
present.
Morphological characteristics. (i) Aerial mycelium.
On ISP
medium
2,
ISP
medium
3,
and ISP medium 4, all supple-
mented with
5
to 10% NaCl, well-developed branched aerial
hyphae are present. The aerial hyphae are white, long, and at
the beginning of sporulation more or less zig-zag shaped
(Fig. 1). The zig-zag-shaped hyphae subdivide into smaller
spores (Fig. 1). Spores are elongated and have smooth
surfaces (Fig.
1).
(ii) Substrate mycelium.
Strain A5-1T produces an exten-
sive substrate mycelium that fragments. It first appears to be
yellowish and then becomes yellowish brown on ISP me-
dium 2, ISP medium
3,
and ISP medium 4.
Physiological characteristics.
Strain A5-iT produces acid
from the following carbohydrates: glucose, inositol, manni-
tol, raffinose, and rhamnose. Acid
is
not produced from
adonitol, dulcitol, erythritol, lactose, melibiose, a-methy1-D-
glucoside, glucitol, and xylose.
Growth does not occur in ISP medium 9 supplemented
with different carbon sources. However, growth occurs in
ISP medium 9 which
is
supplemented with
5
to 10% NaCl
and contains mannose, glucose, rhamnose, maltose, manni-
tol, D-fructose, sucrose, raffinose, glycerol, trehalose, and
myo-inositol as sole sources of carbon but not in medium
containing L-arabinose, D-xylose, D-galactose, and lactose.
VOL.
43,
1993
NEW ACTINOMYCETE SPECIES
269
TABLE
1.
Differential physiological characteristics of strain and other
Nocurdiopsis
species
Nocardiopsis
Nocardiopsis Nocardiopsis
alba
subsp.
alba
subsp.
alba
DSM 43119 DSM 43845T
Nocardiopsis Nocardiopsis
DSM 43111T DSM 40465T DSM 40297T
prasina
Strain
As.lT
dassonvillei alborubida listen
Characteristic
Decomposition
of:
Adenine
+
ND"
+
+
+
+
Hypoxan thine
+
+ +
+
+ +
Tyrosine
+ +
+
+ +
+
Xanthine
+
+
+ +
+
+
Casein
+
+
+
+
+
ND
Urea
Allantoin
-
-
- -
-
+
+
-
- - - -
Hydrolysis
of
Starch
Esculin
Gelatin
Elastin
Arbutin
ND
+
+
+
-
+
+
+
ND
-
+
+
+
+
-
+
+
+
-
-
+
+
+
+
-
+
+
+
-
-
Decarboxylation
of
Citrate
Malate
Succinate
Acetate
Pyruvate
Propionate
Lactate
Oxalate
Acid production from:
Adoni to1
Dulcitol
Erythritol
Glucose
Inositol
Lactose
Mannitol
Melibiose
a-Me thyl-D-glucoside
Raffinose
Rhamnose
Sorbitol
Xylose
+
+
Resistance
to
lysozyme
- -
- -
- -
Production
of:
Phosphatase
P-Galactosidase
P-Glucosidase
+
-
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
Growth in the presence
of:
5.0%
NaCl
+
10.0%
NaCl
+
+
-
+
-
+
-
+
+
+
+
ND,
not determined.
Citrate, malate, succinate, acetate, pyruvate, and propionate
are decarboqdated, but lactate and oxalate are not. Strain
A5-lT
decomposes adenine, hypoxanthine, tyrosine, and
xanthine and hydrolyzes casein, starch, gelatin, and esculin
but not urea, allantoin, and arbutin. It produces catalase and
phosphatase but neither p-galactosidase nor P-glucosidase.
Melanin pigments are not produced on either
ISP
medium
6
or
ISP
medium
7.
The organism reduces nitrates to nitrites.
Strain
A,-,=
is resistant to lincomycin (128 Fg/ml), peni-
cillin G (128 pg/ml), gentamicin
(64
pg/ml), streptomycin
(64
pg/ml), and neomycin
(4
Fg/ml) but not rifampin (128 pg/ml).
The organism is susceptible to lysozyme and tolerates NaCl
at levels
up
to and including
10%.
Chemotaxonomy.
Hydrolyzed whole cells contain
meso-
diaminopimelic acid. Glucose and arabinose are detected in
whole-cell extracts. Thus, strain has a type I11 cell wall
and a variant of the type
C
sugar pattern. Mycolic acids are
not detected. Phospholipid determinations performed with
whole cells indicate that phosphatidylcholine, phosphatidyl-
methylethanolamine, phosphatidylglycerol, and diphos-
270
YASSIN ET
AL.
INT.
J.
SYST.
BACTERIOL.
TABLE 2. Carbon assimilation by strain As-lT and other
Nocardiopsis
spp. on JSP medium 9 supplemented with
5%
NaCl
~~~
Nocardiopsis alba Nocardiopsis Nocardiopsis Nocardiopsis Nocardiopsis alba
Strain
As.lT
subsp.
prasina alborubida
listen
dassonvillei
subsp.
alba
DSM
43845T DSM
4046ST
DSM
40297T
DSM
43111T
DSM
43119
Carbon
source
~~
L-
Arabinose
D-Mannose
D-Glucose
L-Rhamnose
D-Mannitol
D-Fructose
D-Galactose
Maltose
Sucrose
Lactose
Raffinose
Glycerol
Trehalose
myo-Inositol
D-Xylose
-
-
+
+
+
+
+
+
+
+
+
+
+
-
-
+
+
+
+
+
+
+
+
+
+
+
+
-
-
-
+
+
+
+
+
+
+
+
+
+
-
-
+
+
-
-
+
+
+
+
+
+
+
+
+
-
-
+
-
-
phatidylglycerol are present. Therefore, strain
A5-1T
has a
type
PI11
phospholipid pattern. The menaquinones detected
in strain are octahydrogenated menaquinones with 10
isoprene units [MK-lO(H,)]. Minor amounts of tetra-, hexa-,
and decahydrogenated menaquinones with 10 isoprene units
[MK-lO(H,), MK-lO(H,), MK-10(Hl,)], as well as tetra-,
hexa-, octa-, and decahydrogenated menaquinones with nine
isoprene units [MK-9(H4), MK-9(H6), MK-9(H8), MK-
9(Hlo)], are also present. The fatty acid profile
of
strain
consists
of
major amounts of saturated branched-chain fatty
acids
of
the
is0
and anteiso types (iso-CI6:,,
32%;
anteiso-
C17:o, 14.5%; iso-Cl8:,, 9.4%), straight-chain saturated fatty
acids (lt-C16:,, 3.9%; 1.5%;
~Z-C~~:~,
9.9%), and
10-methyl branched-chain fatty acids represented by tuber-
culostearic acid (10-methyl
C18:-,,
15.4%).
DNA-DNA
hybridization.
The renaturation rates as deter-
mined by the optical method reveal binding values that range
from 40 to 46% between strain
A5-1T
and the other
Nocar-
diopsis
species (Table
3).
Type
strain.
The type strain is strain
A5-1
(=
DSM 44048).
It was isolated from soil collected in Alicante, Spain. The
species description is based on
a
single strain and thus serves
as the type strain description.
Another variant, strain
A4-2,
which was found to be
phenotypically identical to
A5-lT,
exhibited a DNA related-
ness level
of
39% to
A5-lT
(Table
3).
However, we hesitate to
suggest a new species for this strain because it is recom-
mended that “a distinct genospecies that cannot be differ-
entiated from another genospecies on the basis of any known
TABLE 3. Degrees
of
DNA binding between and other
Nocardiopsis
species
Organism Degree
of
binding
(%)
to:
4-2
A5-lT 100 39
2
3“
A4-2 39
2
3 100
Nocardwpsis
listen
46 34
2
1
Nocardiopsis alba
subsp.
prasina
42
*
1
44
2
1
Nocardiopsis dassonvillei
44
2
1
65
?
4
Nocardiopsis alborubida
40
*
1
63
2
1
Nocardiopsis alba
subsp.
alba
41
31
-c
1
a
Mean
f
standard deviation.
phenotypic property shall not be named until they can be
differentiated by some phenotypic property” (49).
ACKNOWLEDGMENTS
We thank G. Tuschewitzky (Hygiene Institut Bonn) for preparing
the electron micrograph. E.A.G. thanks G. Vobis (Hoechst A.G.)
for helpful advice during this study.
REFERENCES
1.
Abyzov,
S.
S.,
S.
N. Philipova, and
V.
D. Kuznetsov.
1983.
Nocardiopsis antarcticus,
a new species of actinomycetes,
isolated from the ice sheet of the central antarctic glacier.
Izv.
Akad. Nauk SSSR Ser. Biol. 4559-568.
2.
Athalye, A.,
M.
Goodfellow,
J.
Lacey, and
R.
P.
White.
1984.
Numerical classification
of
Actinornadura
and
Nocardiopsis.
Int.
J.
Syst. Bacteriol. 35:86-98.
3.
Becker,
B.,
M. P. Lechevalier, R. E. Gorden, and
H.
A.
Lechevalier.
1964. Rapid differentiation between
Nocardia
and
Streptornyces
by paper chromatography
of
whole cell hydroly-
sates. Appl. Microbiol. 12:421-423.
4.
Brocq-Rousseau,
D.
1904. Sur un
Streptothrix.
Rev. Bot.
16:
219-230.
5.
Cashion,
P.,
M.
A. Holder-Franklin, J. McCully, and M. Frank-
lin.
1977. A rapid method for the base ratio determination
of
bacterial DNA. Anal. Biochem. 81:461-466.
6.
Collins,
M.
D., T.
Pirouz,
M. Goodfellow, and
D.
E. Minnikin.
1977. Distribution
of
menaquinones in actinomycetes and
corynebacteria.
J.
Gen. Microbiol. 100:221-230.
7.
Cross,
T.,
and R.
W.
Attwell.
1974. Recovery of viable thermo-
actinomycete endospores from deep mud cores, p- 11-20.
In
A. N. Barker, G. W. Gould, and
J.
Wolf (ed.), Spore research.
Academic Press, London.
8.
De Ley,
J.,
H.
Cattoir, and A. Reynaerts.
1970. The quantitative
measurement of DNA hybridization from renaturation rates.
Eur.
J.
Biochem. 12:133-142.
9.
Dittmer,
J.
C.
F.,
and R.
L.
Lester.
1964.
A
simple specific spray
for the detection
of
phospholipids
on
thin-layer chromatograms.
J.
Lipid Res. 5:126-127.
10.
Erikson, D.
1935. The pathogenic aerobic organisms
of
the
Actinomyces
group. Med. Res. Counc. (G.B.) Spec. Rep. Ser.
203: 1-61.
11.
Escara,
J.
F.,
and
J.
R. Hutton.
1980. Thermal stability and
renaturation of DNA in dimethyl sulfoxide solutions: accelera-
tion of the renaturation rate. Biopolymers 19:1315-1327.
12.
Gauze,
G.
F., T.
P.
Preobrazhenskaya, E.
S.
Kudrina, N.
0.
Blinov,
I.
D. Ryabova, and M.
A.
Sveshnikova (ed.).
1957.
Problems of classification of
actinomycetes-antagonists.
Gov-
ernment Publishing House of Medical Literature, Moscow.
VOL. 43, 1993 NEW ACTINOMYCETE SPECIES 271
13.
Gauze, G. F., T.
S.
Maksimova,
0.
L.
Olkhovatova, M. A.
Sveshnikova, G.
V.
Kochetkova, and G.
B.
Ilchemko.
1974.
Production
of
madumycin, an antibacterial antibiotic, by
Acti-
nornadura flava
sp. nov. Antibiotiki (Moscow) 9:771-775.
14.
Gauze, G.
F.,
G.
V.
Sveshnikova,
R.
S.
Ukholina, G. N.
Komorova, and
V.
S.
Bashanov.
1977. Production
of
nocamycin,
a new antibiotic, by
Nocardiopsis syringae
sp. nov. Antibiotiki
(Moscow) 22:483436.
15.
Gordon,
R.
E.
1966. Some criteria for the recognition of
Nocardia madurae
(Vincent) Blanchard. J. Gen. Microbiol.
45355-364.
16.
Gordon,
R.
E.
1967. The taxonomy
of
soil bacteria, p. 239-321.
In
T. R. G. Gray and D. Parkinson (ed.), The ecology of soil
bacteria. Liverpool University Press, Liverpool, United King-
dom.
17.
Gordon,
R. E.,
and D.
A.
Barnett.
1977. Resistance to rifampin
and lysozyme
of
strains
of
some species of
Mycobacterium
and
Nocardia
as a taxonomic tool. Int. J. Syst. Bacteriol. 27:176-
178.
18.
Gordon,
R.
E.,
and
A.
C. Horan.
1968.
Nocardia dassonvillei,
a
macroscopic replica of
Streptomyces griseus.
J. Gen. Micro-
biol. 50:235-240.
19.
Gordon,
R.
E.,
and
J.
M. Mihm.
1962. The type species
of
the
genus
Nocardia.
J. Gen. Microbiol. 27:l-10.
20.
Grund,
E.,
and
R.
M. Kroppenstedt.
1989. Transfer of five
Nocardiopsis
species to the genus
Saccharothrix
Labeda et al.
1984. Syst. Appl. Microbiol. 12:267-274.
21.
Grund,
E.,
and
R.
M. Kroppenstedt.
1990. Chemotaxonomy and
numerical taxonomy of the genus
Nocardiopsis
Meyer 1976.
Int. J. Syst. Bacteriol. 405-11.
22.
HUSS,
V. A.
R.,
H. Festl, and K.
H.
Schleifer.
1983. Studies on
the spectrophotometric determination of DNA hybridization
from renaturation rates. Syst. Appl. Microbiol. 4:184-192.
23.
Jahnke, K.-D.
1992. Basic computer program for evaluation of
spectroscopic DNA renaturation data from GILFORD
SYS-
TEM 2600 spectrophotometer on a PC/XT/AT type personal
computer. J. Microbiol. Methods 1961-73.
24.
Kroppenstedt,
R.
M.
1992. The genus
Nocardiopsis,
p. 1139-
1156.
In
A. Balows, H.
G.
Truper, M. Dworkin, W. Harder, and
K.
H.
Schleifer (ed.), The prokaryotes, 2nd ed. Springer Verlag,
New York.
25.
Kroppenstedt,
R.
M.,
E.
Stackebrandt, and
M.
Goodfellow.
1990. Taxonomic revision of the actinomycete genera
Actino-
madura
and
Microtetraspora.
J. Syst. Appl. Bacteriol. Micro-
biol. 13:18&160.
26.
Kurup,
V.
P.,
and
J.
N. Fink.
1975. A scheme for the identifi-
cation of thermophilic actinomycetes associated with hypersen-
sitivity pneumonitis. J. Clin. Microbiol. 255-61.
27.
Kurup,
V.
P., and
J.
A.
Schmitt.
1973. Numerical taxonomy of
Nocardia.
Can. J. Microbiol. 19:1035-1048.
28.
Labeda, D. P., and M. P. Lechevalier.
1989. Amendment of the
genus
Saccharothrix
Labeda et al. 1984 and descriptions
of
Saccharothrix espanaensis
sp. nov.,
Saccharothrix cryophilis
sp. nov., and
Saccharothrix mutabilis
comb. nov. Int. J. Syst.
Bacteriol. 3k420-423.
29.
Labeda, D.
P., R.
T. Testa,
M.
P. Lechevalier, and H. A.
Lechevalier.
1984.
Saccharothrix,
a new genus
of
the
Actinomy-
cetales
related to
Nocardiopsis.
Int. J. Syst. Bacteriol. 34:426-
431.
30.
Lechevalier, H.
A.,
and M. P. Lechevalier.
1970. A critical
evaluation
of
the genera
of
aerobic actinomycetes, p. 393-405.
In
H. Prauser (ed.), The
ActinomycetaZes.
VEB Gustav Fischer
Verlag, Jena, Germany.
31.
Lechevalier, M. P.
1968. Identification of aerobic actinomycetes
of clinical importance.
J.
Lab. Clin. Med. 71:934-944.
32.
Lechevalier, M.
P.,
C. de Bievere, and H. A. Lechevalier.
1977.
Chemotaxonomy
of
aerobic actinomycetes: phospholipid com-
position. Biochem. Syst. Ecol. 5249-260.
33.
Liegard, H., and M. Landrieu.
1911. Un cas de mycose con-
junctivale. Ann. Ocul. 14tk418-426.
34.
Mesbah, M., U. Premachandran, and W. B. Whitman.
1989.
Precise measurement
of
the G+C content
of
deoxyribonucleic
acid by high-performance liquid chromatography. Int. J. Syst.
Bacteriol. 39:159-167.
35.
Meyer, J.
1976.
Nocardiopsis,
a
new genus of the order
Acti-
nomycetales.
Int. J. Syst. Bacteriol. 26:487-493.
36.
Meyer,
J.
1989. Genus
Nocardiopsis
1976, p. 2562-2569.
In
S.
T. Williams, M.
E.
Sharpe, and J.
G.
Holt (ed.), Bergey’s
manual of systematic bacteriology, vol. 4. Williams and
Wilkins, Baltimore.
37.
Minnikin, D.
E.,
L. Alshamaony, and
M.
Goodfellow.
1975.
Differentiation
of
Mycobactenurn, Nocardia
and related taxa by
thin-layer chromatographic analysis of whole-cell methanoly-
sates. J. Gen. Microbiol. 88:200-204.
38.
Minnikin, D.
E.,
I.
G.
Hutchinson, A.
B.
Caldicott, and M.
Goodfellow.
1980. Thin-layer chromatography of methanoly-
sates
of
mycolic acid-containing bacteria.
J.
Gen. Chromatogr.
188:221-233.
39.
Minnikin,
D.
E.,
A. G. O’Donell, M. Goodfellow, G. Alderson,
M. Athalye, A. Schaal, and
J.
H. Parlett.
1984. An integrated
procedure for the extraction
of
isoprenoid quinones and polar
lipids. J. Microbiol. Methods 2:233-241.
40.
Miyashita,
M.,
Y.
Mikami, and T. Arai.
1984. Alkalophilic
actinomycete,
Nocardiopsis dassonvillei
subsp.
prasina
subsp.
nov., isolated from soil. Int.
J.
Syst. Bacteriol. 34405409.
41.
Preobrazhenskaya, T. P., and M. A. Sveshnikova.
1974. New
species
of
the genus
Actinomadura
.
Mikrobiologiya 43:864-868.
42.
Preobrazhenskaya, T.
P.,
M.
A. Sveshnikova, and G. F. Gauze.
1982.
On
the transfer of certain species
of
the genus
Actinoma-
dura
Lechevalier et Lechevalier 1970 to the genus
Nocardiopsis
Meyer 1976. Mikrobiologiya 51:lll-113.
43.
Shearer, M. C., P. M. Colman, and C.
H.
Nash 111.
1983.
Nocardiopsis mutabilis,
a new species of nocardioform bacteria
isolated from soil. Int. J. Syst. Bacteriol. 33:369-374.
44.
Shirling,
E.
B.,
and D. Gottlieb.
1966. Methods for character-
ization
of
Streptomyces
species. Int. J. Syst. Bacteriol. 16:313-
340.
45.
Takahashi, A., K. Hotta,
N.
Saito, M. Morioka,
Y.
Okami, and
H. Umezawa.
1986. Production
of
novel antibiotic, dopsisamine,
by a new subspecies
of
Nocardiopsis mutabilis
with multiple
antibiotic resistance. J. Antibiot. 39:175-183.
46.
Tsukamura, M.
1974. Differentiation
of
the
“Mycobacterium”
rhodochrous
group from
Nocardia
by p-galactosidase activity.
J. Gen. Microbiol. 80:553-555.
47.
Wagner, H., L. Horhammer, and
L.
Wollf.
1961. Dunnschicht-
chromatographie von Phosphatiden und Glycolipiden. Bio-
chem.
Z.
334:175-184.
48.
Waksman,
S.
A,
and
A.
T. Henrici.
1948. Family
Actinomyce-
taceae
Buchanan and family
Streptomycetaceae
Waksman and
Henrici, p. 961.
In
R.
S.
Breed,
E.
G.
D.
Murray, and
A.
P.
Hitchens (ed.), Bergey’s manual
of
determinative bacteriology,
6th ed. The Williams and Willkins Co., Baltimore.
49.
Wayne,
L.
G., D.
J.
Brenner,
R.
R.
Colwell, P.
A.
D. Grimont,
0.
Kandler, M.
I.
Krichevsky,
L.
H. Moore, W.
E.
C.
Moore,
R.
G.
E.
Murray,
E.
Stackebrandt, M. P. Starr, and
H.
G.
Triiper.
1987. Report of the Ad Hoc Committee on Reconcilia-
tion of Approaches to Bacterial Systematics. Int. J. Syst.
Bacteriol. 37:463-464.
