INTERNATIONAL JOURNAL O F
SYSTEMATIC BACTERIOLOGY
Vol. 18, No. 3
July 1968
pp. 207-229
Copyright 1968 by the Iowa State University P r e s s
A COMPARISON O F 120 STRAINS O F BACTERIUM
ANITRATUM SCHAUB AND HAUBER WITH THE
T Y P E STRAIN O F THIS SPECIES
Rudolph Hugh and Robert Reese*
The George Washington University School of Medicine,
Department of Microbiology, Washington, D. C.
ABSTRACT. M o r e t h a n 5 0 c h a r a c t e r i s t i c s f o r
e a c h of 1 2 0 s t r a i n s of B a c t e r i u m a n i t r a t u m
Schaub and Hauber 1948 w e r e c o m p a r e d with
t h e c h a r a c t e r i s t i c s o f t h e t y p e s t r a i n o f B_.
anitraturn, American Type Culture Collection
( A T C C ) 1 9 6 0 6 , t o e x p l o r e t h e p a r a m e t e r s of
the strains.
T h e i n c i d e n c e of e a c h c h a r a c t e r
among t h e 120 s t r a i n s is e x p r e s s e d quantitatively.
T h e s e s t r a i n s of t h e s p e c i e s h a v e a
definite reproducible biochemical reaction
p a t t e r n by which t h e s p e c i e s m a y be expediently recognized.
B. a n i t r a t u m i s a v a l i d l y
published illegitimate name according to the
I n t e r n a t i o n a l C o d e of N o m e n c l a t u r e of B a c teria.
The oldest known available l e g i t i m a t e
name for this species appears to be Achromobacter anitratus (Schaub and Hauber) Bourlat
m 4 3 .
D e B o r d ' s ( 1 9 4 2 ) d e s c r i p t i o n of
H e r e l l e a v a g i n i c o l a d o e s n o t f i t a n y of t h e 1 2 0
s t r a i n s o f ,B. a n i t r a t u m s t u d i e d . D i p l o c o c c u s
m u c o s u s v o n L i n g e l s h e i m 1 9 0 6 a n d FJ. v a g i n i c o l a a r e n o t s e n i o r s y n o n y m s f o r 2. a n i t r a t u m .
T h e p r o p o s e d n e o t y p e s t r a i n of A c h r o m o b a c t e r
m u c o s u s , t h e t y p e s t r a i n s o f ,A. c o n j u n c t i v a e ,
a n d A. h a e m o l y t i c u s s u b s p . h a e m o l y t i c u s a r e
None
i d e n t i f i e d a s s t r a i n s o f 2. a n i t r a t u m .
of t h e s e 3 n a m e s i s a s e n i o r s y n o n y m o f B,.
anitratum.
-
* Present address:
Yale University, Department of Microbiology, New Haven, Connecticut.
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INTERNATIONAL
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INTRODUCTION
=-
Schaub and Hauber (1948) isolated 15 s t r a i n s of a v e r y
distinctive schizomycete which they named B a c t e r i u m
tratum.
It i s a Gram-negative nonmotile rod.
The type
s t r a i n of t h i s s p e c i e s is ATCC 19606(Hugh and R e e s e 1967).
The objective of t h i s r e p o r t is to c o m p a r e the c h a r a c t e r i s t i c s of 120 s t r a i n s of ,B. anitratum, isolated f r o m d i v e r s e
s o u r c e s , in a n attempt t o d e t e r m i n e the p a r a m e t e r s of the
s t r a i n s . This study was initiated in a n attempt to demons t r a t e the d e g r e e of variation encountered among putative
s t r a i n s of t h e s p e c i e s and to alleviate c e r t a i n problems of
recognition which have o c c u r r e d .
Reference s t r a i n s of
various schizomycetes w e r e studied while pursuing the above
objectives. Among t h e s e r e f e r e n c e s t r a i n s w e r e the p r o posed neotype of Achromobacter mucosus and the type s t r a i n s
of A. conjunctivae and A. haemolyticus subsp. haemolyticus.
T h e purpose of t h i s r e p o r t was extended to c o m p a r e t h e s e 3
nomenclatural types with the type s t r a i n of B. a n i t r a t u m .
Some of the s n a r l e d nomenclatural problems concerning t h i s
taxon will be discussed.
MATERIALS AND METHODS
The type s t r a i n of B. a n i t r a t u m was isolated f r o m the
u r i n e of a patient with a u r i n a r y t r a c t infection and was
among the 15 original s t r a i n s studied by Schaub and Hauber
(1948). This culture was obtained f r o m the National Communicable D i s e a s e Center, Atlanta, Georgia, in 1965, where
it was deposited in 1949 by Schaub, and a descendant ( R H
2208) was sent to ATCC i n 1966 where it was accessioned
a s s t r a i n 19606.
The proposed neotype s t r a i n of Achromob a c t e r mucosus and the 2 type s t r a i n s f o r A. conjunctivae
and A. haemolyticus subsp. haemolyticus w e r e obtained f r o m
ATC?.
These 4 type and proposed neotype s t r a i n s a r e
m a r k e d with a s t e r i s k s in Table 1. The 120 s t r a i n s of B.
anitratum w e r e isolated f r o m f r o z e n peas and m e a t i n a
"television dinner," f r o m soil, animals, and man.
The
s t r a i n s of human origin w e r e isolated f r o m the r e s p i r a t o r y
t r a c t (sputa, lungs, oropharyngeal swabs), c e r e b r o s p i n a l
fluid, blood, u r i n e , conjunctiva, skin, wounds, e a r , and abc e s s e s . Eleven of the 1 2 0 s t r a i n s w e r e isolated by Isabelle
Schaub and a t l e a s t 4 of t h e s e w e r e used in h e r 1948 r e p o r t .
Over 50%of the 120 s t r a i n s w e r e received as H e r e l l e a vaginicola o r H e r e l l e a sp. O t h e r s w e r e received a s various
s p e c i e s of Achromobacter, Acinetobacter, Alcaligenes, Bat-
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SYSTEMATIC
BACTERIOLOGY
2 09
--
t e r i u m , Cytophaga, Diplococcus, Micrococcus, Mima, Moraxella, and N e i s s e r i a . The names and corresponding a c c e s sion numbers of 65 s t r a i n s which historically may contribute
to a b e t t e r understanding of t h e taxon a r e listed in Table 1.
The procedures used to study the above o r g a n i s m s have
been described by Hugh and R e e s e (1967).
RESULTS
The c h a r a c t e r i s t i c s of t h e 4 type and neotype s t r a i n s a r e
recorded adjacent to each other in Table 2 to facilitate d i r e c t
comparison. The c h a r a c t e r i s t i c s of 1 2 0 s t r a i n s of B. anit r a t m a r e a l s o r e c o r d e d in Table 2 to expedite comparison
with t h e type s t r a i n of B. anitratum. S e v e r a l of the c h a r a c t e r i s t i c s of the 120 s t r a i n s , for various r e a s o n s , w a r r a n t
elaboration and special emphasis.
The morphology of the s t r a i n s varied f r o m distinctly
coccoid f o r m s and rod-shaped cells to filaments. They w e r e
Gram-negative, asporogenous, and nonmotile. The s t r a i n s
w e r e not examined f o r gliding o r twitching surface t r a n s location. These s t r i c t a e r o b e s produced a dense turbidity
in n e u t r a l peptone broth and b r a i n - h e a r t infusion broth in
18-24 hours at 30 and 37°C. Most of the s t r a i n s produced
abundant, smooth, glistening, 0 . 5 to 2 mrn d i a m e t e r colonies
on infusion a g a r a f t e r 24 hours of incubation at 30°C. A
c h a r a c t e r i s t i c unpleasant odor was produced by m o s t s t r a i n s .
Water- soluble pigments (pyocyanine and pyoverdine) and
indophenol oxidase w e r e not produced,
The s i z e and number of colonies on desoxycholate a g a r
was often greatly reduced when compared with t h e growth
response on infusion agar. Some s t r a i n s produced white o r
translucent colonies on de soxycholate a g a r . The d i s c r e t e
colonies of other s t r a i n s appeared pink whereas colonies in
a r e a s of dense growth appeared white when. observed a f t e r
24 hours of incubation on desoqrcholate a g a r . This medium
did not support the growth of a l l the s t r a i n s studied. Abundant bluish-gray colonies w e r e s e e n on H e r e l l e a Agar (Difco
0909, Mandel et
al. 1964) a f t e r 24 hours of incubation at
- 30°C. The colony color apparently was due to absorption of
b r o m o c r e s o l purple f r o m t h e medium.
The 120 s t r a i n s failed to grow and did not produce acid
in sealed tubes of O F medium containing glucose. All the
s t r a i n s promptly produced acid in open tubes of O F medium
Eighty-three percent of the
containing glucose o r xylose.
s t r a i n s produced acid within 24 hours in open tubes of O F
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INTERNATIONAL JOURNAL
210
T a b l e 1.
S i x t y - f i v e p e r t i n e n t s t r a i n s of B a c t e r i u m a n i t r o t u m a n d t h e i r
corresponding strain numbers.
Received a s
RH
ATCC
A l c a l i g e n e s sp.
Achromobacter anitratum
24
461
2378
46 3
46 5
467
47 3
47 3
17 5
47 6
47 7
478
47 9
2207
2208
2197
10153
17912
17913
19649
231 5
19568
9214
2316
19 569
9213
2317
19 187
9212
2314
19682
9211
483
485
1935
19 36
1937
2204
9955
9956
11959
15567
I5566
15568
14682
2210
221 1
221 2
17903
8102
1790eC*10303
17905* 10304
Billing A l ; ST 1
Billing A3; S T 3
Billing B 4
Billing B 10
Billing B 9
King 2566
King 2656
King 2673
King 2713
King 2714
King 2734
King 2744
King S c h a u b 77
King S c h a u b 8 1
Schaub Biol. 1 ;
F e r g u s o n 5W101
Schaub Biol. 2;
S n e a t h D. 409
S c h a u b 90; S n e a t h D.
410
S c h a u b 9 3 ; S n e a t h D.
41 I
S n e a t h D. 408; Eddy;
i s o l a t e d by S c h a u b
Deacon 6-561
Deacon 5- 156A
Cilman
E l l i s 1972 a n d 5209
E l l i s 2409 a n d 5345
E l l i s 11208 a n d 4744
CDC KC 138; S e e l i g e r
V 2240-52
Stuart A 267
S t e n z e l 3516/60
S t e n z e l P544/60
221 3
17906* 10305
S t e n z e l 2446/60
17910
19679
S t e n z e l P790/60
F e r g u s o n 5W69S t u a r t H 1 168
F e r g u s o n 5W71S t u a r t H 1 166
F e r g u s o n 5W79
S t u a r t H 1169
F e r g u s o n 5W81
S t u a r t H 1167
-~
Bacterium anitratum
H e r e l l e a "p.
H e r e l l e a sp.
Bacterium anitratum
H e r e l l e a sp.
484
Bacterium anitratum
Diplococcus m u c o s u s
Acinetobacter anitratus
Achromobacter mucosus
Achromobacter conjunctine
A c h r o m o b a c t e r haernolyticus
subsp. h a e m o l y t i c u s
19678
19606"
15 308
2226
15149
2227
19680
2228
19681
NCTC
NCIB
Other
FDA P C I - 3
9115
9116
--
7814
10309
9 299
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---
Schaob
Schaub
Schaub
Schaub
SYSTEMATIC
BAC TERIO LOGY
21 1
T a b l e 1 continued
RH
Received a s
ATCC
H e r e l l e a vaginicola
2264
H e r e l l e a sp.
1429 3
14290
14294
17961
17922
14987
Acinetobacter a n i t r a t u n
Cytophaga a n i t r a t a
2421
2418
2422
2424
2382
2338
2412
241 3
2380
2385
Diplococcus m u c o s u s
2386
17957
H e r e l l e a vaginicola
2390
2425
2392
2287
2270
227 1
2272
19003
19004
17979
15473
2273
19684
H e r e l l e a vaginicola
Acinetobacter winogradskyi
Micrococcus cerificans
M o r a x e l l a glucidolytica
H e r e l l e a sp.
H e r e l l e a vaginicola
227 4
227 5
2276
2277
2278
2279
2280
228 1
2282
*
-
**
17924
17945
19683
NCTC
NCIB
9017
9019
Other
-
zt
C a r y 18300 Daly
51.
Case 2
C a r y 499
CDC 1847
CDC 1856
CDC 7788; K a a s 15860
B r i s o u 12
Kallio H.O. 1
Kallio H. 0. 3
Kallio H. 0. 4
Brisou 64
Lautrop A l ; Stuart
B5W-3777
L a u t r o p A194;
S e e l i g e r D5/62
C o u r t i e u 57.07 1.066
C o u r t i e u 57.07 1.228
P i e c h a u d A165
FDA PCI-1788; Chang
Nelson N1134 and 5
N e l s o n N1138 a n d 6
N e l s o n N1140 a n d 7King 7411
N e l s o n N1141 and 8King 680 1
Nelson N1142 and 9King 749 5
N e l s o n N1143 and 10King 7435
Nelson N1175 and 14
Nelson N1176 and 15
N e l s o n N1177 and 16
N e l s o n N1180 and 19
N e l s o n N1181 and 20
N e l s o n N1187
N e l s o n N1244 a n d 50
type strain
p r o p o s e d neotype
received f r o m
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JOURNAL
medium (Hugh and Leifson 1953, Difco 0688) containing 1%
l a c t o s e ; 98'70 produced oxidative acidity f r o m 170 l a c t o s e
within 48 h o u r s .
About 65% of t h e s t r a i n s g r e w and produced oxidative
acidity f r o m glucose in the p r e s e n c e of 0.001 m o l a r iodoa c e t a t e ; the remaining s t r a i n s failed t o g r o w i n t h e p r e s e n c e
of t h i s concentration of iodoacetate.
Many of t h e s t r a i n s
produced acid f r o m ethanol.
A v e r y c l e a r zone of haemolysis developed around d i s c r e t e s u r f a c e colonies of 1370of the s t r a i n s growing on 3. 5%
defibrinited rabbit blood a g a r .
Haemodigestion often appeared around confluent s u r f a c e growth on blood a g a r . Two
s t r a i n s which caused l a r g e c l e a r zones of h a e m o l y s i s around
d i s c r e t e s u r f a c e colonies w e r e studied in g r e a t e r detail and
w e r e found to produce f i l t e r a b l e heat-labile haemolysin.
The d i s c r e t e zone of haemolysis was readily recognized and
distinguished f r o m haemodigestion on rabbit blood a g a r when
the medium contained 3- 10 colonies distributed o v e r onehalf of the p e t r i dish a r e a . Sheep and human e r y t h r o c y t e s
generally w e r e m o r e r e s i s t a n t to haemolysis then rabbit
c e l l s ; the zones of haemolysis i n sheep and human blood
a g a r generally did not a p p e a r until the second dayof incubation and the zones of h a e m o l y s i s w e r e often only p a r t l y
cleared.
DISCUSSION
B. a n i t r a t u m h a s a v e r y p r e c i s e biochemical r e a c t i o n
p a t t e r n by which s t r a i n s of the s p e c i e s readily can be d i s B. -anitinguished f r o m o t h e r schizomycetes. The ability of t r a t u m t o produce prompt acidity f r o m glucose and l a c t o s e
i n t h e p r e s e n c e of air and the inability t o produce acid f r o m
t h e s e s u g a r s i n t h e a b s e n c e of a i r was noted by Hugh and
Leif son ( 19 5 3 ) . T h i s p a t t e r n of activity distinguishes t h i s
s p e c i e s f r o m nonmotile anaerogenic slow l a c t o s e fermenting
and l a c t o s e nonfermenting Enterobacte riaceae.
Twenty-four strains of B. a n i t r a t u m w e r e studied m o r e
than 10 y e a r s ago (unpublished data f r o m t h e files of t h e
s e n i o r author). T h e s e 2 4 s t r a i n s w e r e restudied and included
among the 120 strains of _B. a n i t r a t u m d e s c r i b e d i n t h i s r e port.
The a t t r i b u t e s studied 10 y e a r s ago included G r a m
reaction, c e l l shapk, s p o r e formation, motility, production
of acid in O F medium containing a r a b i n o s e , glucose (open
and sealed), fructose, galactose, l a c t o s e (open and sealed),
maltose, mannitol, mannose, rhamnose, s u c r o s e and xylose.
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SYSTEMATIC BACTERIOLOGY
T a b l e 2.
21
T h e c h a r a c t e r i s t i c s of t h e t y p e s t r a i n of B a c t e r i u m a n i t r a t u m (RH E208).
A c h r o m o b a c t e r m u c o s u s ( R H 221 l ) , A c h r o m o b a c t e r c o n j u n c t i v a e ( k H 2212),
A c h r o m o b a c t e r h a e m o l y t i c u s s u b s p . h a e m o l y t i c u s ( R H 221 3), H e r e t l e a v a g i n i c o l a t ,
a n d 120 s t r a i n s of B a c t e r i u m a n i t r a t u m .
RH S t r a i n N u m b e r s
4H.
2208 2211 2212 2213 vag.
G r a m negative, rod-shaped
Spores, motility
G r o w t h a t 22, 30 a n d 37'C
G r o w t h a t 42'C
F'yocyanine p r o d u c t i o n
t
t
t
t
t
t
t
t
Strict aerobe
Adonitol
1-Arabinose
C e 110b i o s e
G l u c o s e open
t
Glucose sealed, glucose g a s
Dulcitol (dulcite)
d-Fructose
d-Galactose
G l y c e r o l , 1-Inositol, h u l i n
Lactose
Maltose
d-Mannitol (mannite)
d-Mannose
Melezitose
Melibiose
Raffinose
Rhamnose
d - Ribose
Salicin, Sucrose
d-Sorbitol, T r e h a l o s e
d-Xylose
Indole
Methyl r e d
Acetylmethyl carbinol
Gitrate. Simmons
Citrate, Christensen
Catalase
Indophenol o x i d a s e
Gelatin, charcoal
H y d r o g e n s u l f i d e , TSI
2-Ketogluconate
t
t
t
t
t
+
t
t
t
120 S t r a i n s of B. a n i t r a t u r n
t
120
0
120
103
0
t
120
t
t2
t
118
0
t
0
t5.
t,o
0
100
0
100
0
100
0
100
100
100
0
0
120
0
0
0
0
0
0
0
0
0
120
0
0
0
0
0
0
0
1 a0
12b
120
0
120
100
2
117
2
0
118
16
0
120
103
0
0
17
120
t
+I1
t
t
t
t
t
t
tz
t
+L
t
+I4
t
0
1
118
t
t
t
0
119
0
120
0
120
0
t
t
102
104
t
t
t
+28
+I
120
0
I
0
0
86
0
0
0
0
100
0
100
86
0
0
0
120
0
0
10
0
100
0
0
109
0
119
0
0
0
0
120
1
0
120
120
0
0
99
100
0
0
100
I20
0
0
45
120
0
0
120
0
t
t
120
0
0
0
0
0
t
t
%t
0
120
0
17
120
119
+7
t5
-
0
I20
119
99
120
t
+L
t1
I20
120
15
11
0
105
109
4
12
0
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16
0
120
104
120
1 LO
0
88
91
100
0
13
0
0
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T a b l e 2 continued
RH S t r a i n N u m b e r a
*H.
2208 2 2 1 1 2 2 1 2 2 2 1 3 vag.
-
-
-
-
-
t
-
-
_
-
Lvsinc decarboxylase, ninhydrin
Malonate
Nitrate to gas
N i t r a t e to n i t r i t e
Zinc dust test on negative
nitrite tests
+
+
Phenylalanine deaminase
-
-
G r o w t h in p o t a s s i u m c y a n i d e b r o t h
l:rea, Christensen
Extracellular deoxyribonuclease
H a c m o l y s i s . r a b b i t blood
i
-
-
t
-
-
l
_
-
i
_
-
-
t
t
-
_
+
120 S t r a i n s of
ia
ts
0
61
_
0
-
-
-
*
t
t
t,,
76
0
0
I LO
t
_
B. a n i t r a t u m
0
25**
74
1
15
-
%-
I20
49
I20
120
0
63
0
100
I20
0
28
0
0
34
89
86
31
74
1
16
119
104
0.8
13
Positive
Negative
N o m e n c l a t u r a l t y p e ( d e s c r i p t i o n ) r e c o r d e d by Dr B o r d 1942. T a b l e 2, g r o u p g. p. 474.
**
G r o w t h u s u a l l y m e a g e r when c o r n p a r c d t o t h a t of K l e b s i e l l a p n e u m v n i a e .
S u b s c r i p t Day o n which the r e a c t i o n b e c a m e p o s i t i v e .
t
-
t.
~-
The following t e s t s w e r e a l s o performed: indole, methyl r e d ,
acetylmethyl carbinol, c i t r a t e (Simmons), c a t a l a s e , indophenol oxidase, gelatin liquefaction, hydrogen sulfide (Kligl e r ) , n i t r a t e reduction t o nitrogen gas, n i t r a t e reduction to
nitrite, zinc dust t e s t t o d e t e r m i n e t h e p r e s e n c e of u n r e duced nitrate, u r e a (Christensen), haemolysis. around dis Crete s u r f a c e colonies on rabbit blood a g a r . The a t t r i b u t e s
of the 24 s t r a i n s , r e c o r d e d during studies p e r f o r m e d 10
y e a r s ago, w e r e t h e s a m e a s the c h a r a c t e r i s t i c s of descendants of t h e s e s t r a i n s r e c o r d e d during studies c a r r i e d out
for this r e p o r t .
This comparative study r e f l e c t s t h e stable
n a t u r e of the s p e c i e s and t h e validity of t h e u s e of t h e s e att r i b u t e s t o recognize the s p e c i e s .
A Gram-negative, atrichous, nonmotile, asporogenous,
coccoid o r rod- shaped schizomycete capable of growing a t
30 to 3 7 ° C on nutrient a g a r , and which does not produce
indophenol oxidase, produces oxidative acidity f r o m glucose
but fails to f e r m e n t glucose anaerobically should be p r e sumptively identified as ,B. anitratum.
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BAG T E R I O L O G Y
215
The need for dividing the taxon described in Table 2 into
2 o r m o r e species o r v a r i e t i e s based on carbohydrate r e actions, gelatinolytic activity, haemo ly s i s, animal patho genicity, o r s o u r c e of isolation does not a p p e a r w a r r a n t e d
at t h i s time; t h i s kind of subdivision should be considered
infrasubspecific.
The taxon probably should not be divided
into subgroups on the b a s i s of t h e u r e a s e reaction, growth
r e s p o n s e in broth containing potassium cyanide o r malonate
reaction since numerous s t r a i n s produced t e s t reactions
which in our hands and according to the technique employed
w e r e often equivocal and difficult to i n t e r p r e t even when
compared with suitable controls.
Thirteen of the 32 s t r a i n s of motile H e r e l l e a vaginicola
described by Nelson and Shelton (1965) w e r e made available
f o r reexamination (vixe Table 1). We w e r e unable to confirm
motility o r detect flagella on c e l l s of t h e s e s t r a i n s .
None of the 120 s t r a i n s d e s c r i b e d in Table 2 reduced
n i t r a t e t o n i t r i t e when grown i n pancreatic digest of c a s e i n
containing y e a s t extract. Apparently ,B. a n i t r a t u m can exp r e s s n i t r a t e reduction only under c e r t a i n defined conditions.
The nitrogen requirement of B,. a n i t r a t u m i s generally s a t i s fied by ammonium sulfate, S e v e r a l s t r a i n s of ,B. anitratum
have been adapted to grow i n a minimal medium in which
n i t r a t e o r n i t r i t e i s the only nitrogen s o u r c e added. One
s t r a i n of ,B. a n i t r a t u m failed to grow in this minimal medium
without leucine ( J y s s u m and J o n e r 1965). Nitrate is reduced
The biosynthesis
to n i t r i t e by n i t r a t e grown 2. anitrahun.
of the n i t r a t e reducing s y s t e m is r e p r e s s e d by a s p a r t i c
acid and alanine; the activity of t h i s s y s t e m is inhibited by
ammonium ion, a s p a r t i c acid, and asparagine ( J y s s u m and
J o n e r 1966).
The guanine and cytosine in deoxyribonucleic acid of
s e v e r a l s t r a i n s of ,B. a n i t r a t u m h a s been estimated to be
about 40 m o l e s percent (Catlin and Cunningham 1964; S e e s e
e t 51. 1965; Hugh and R e e s e 1967; Bbvre 1967) with a range
of 41.5 to 43 (De Ley et al. 1967).
Schaub Et
(1948) observed that a n a n t i s e r u m p r e p a r e d
by inoculating a rabbit with a n encapsulated s t r a i n of B.
a n i t r a t u m caused the capsuleof all M phase strains to swell.
Mucoid B. a n i t r a t u m s t r a i n s w e r e d e s c r i b e d to be serologically homogeneous.
Unencapsulated, smooth, and rough
v a r i a n t s failed t o show a positive Quellung reaction. Stuart
e t al.
- (19 49) found t h e i r nonmotile s t r a i n s w e r e antigenically
heterogeneous.
F e r g u s o n and R o b e r t s (1950) described 10
capsular antigens among 109 nonmotile B. anitratum s t r a i n s .
al.
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P u r e c a p s u l a r a n t i s e r a w e r e p r e p a r e d by absorption of whole
a n t i s e r a with heterologous o r homologous o r g a n i s m s . Sixtyeight percent of 75 s t r a i n s in the M phase w e r e agglutinated
by one of the 10 c a p s u l a r a n t i s e r a . They c o r r o b o r a t e the
Ewing (1949) and Stuart observation that suspensions of M
phase B. a n i t r a t u m agglutinated poorly o r not a t all a f t e r
heating-at 100°C for 1 hour when exposed to homologous
a n t i s e r a p r e p a r e d with living encapsulated s t r a i n s . The 0
antigens of 2 s t r a i n s w e r e determined.
Deacon's (1945)
s t r a i n s of H e r e l l e a a n d s t r a i n s of B. a n i t r a t u m f r o m F e r guson w e r e agglutinated by B. a n i t r a t u m a n t i s e r u m (Ewing
1949).
-
Gary et al. (1956) d e s c r i b e d 9 s e r o t y p e s (11 through 19)
of Mima polymorpha (not B. anitratum) by using a proteolytic enzyme f o r the preparation of the precipitating antigens. T h e s e 9 M. polymorpha s e r o t y p e s w e r e added to t h e
10 ,B. anitratm-serotypes
described by F e r g u s o n and Robe r t s ; collectively they w e r e d e s c r i b e d a s "19 s e r b t y p e s of
Mima polymorpha."
C a r y e-t al.
- (1958) reported that living
suspensions of s o m e H e r e l l e a sp. a r e inagglutinable in
homologous a n t i s e r a .
They reported 3 group antigens in
Herellea sp. and found it expedient to u s e a n absorbed polyvalent Herellea a n t i s e r u m which was p r e p a r e d by mixing
equal volumes of a n t i s e r a p r e p a r e d f r o m 3 H e r e l l e a s t r a i n s .
The slide agglutination t e s t was p e r f o r m e d with c e l l s which
had been boiled in sodium hydroxide.
T h i s polyvalent antis e r u m agglutinated 38 s t r a i n s of H e r e l l e a sp. but did not
agglutinate 10 _B. a n i t r a t u m s t r a i n s ( 10 s e r o t y p e s obtained
f r o m Ferguson) and 42 Mima polymorpha s t r a i n s . F e r g u s o n ' s 10 s e r o t y p e s of ,B. a n i t r a t u m w e r e agglutinated by a
polyvalent Mima polymorpha a n t i s e r u m .
Seventy-three per cent of 100 H e r e l l e a sp. s t r a i n s isolated f r o m various s o u r c e s other than the blood w e r e typed
with seven capsular a n t i s e r a (Daly z t a1
-.
1962). S t r a i n s
isolated f r o m the blood generally r e a c t e d with one c a p s u l a r
antiserum.
C e l l - f r e e e x t r a c t s of sonically disrupted s t r a i n s received
a s H. vaginicola, Mima polymorpha, B. anitratum, e t c . ,
w e r e studied by Mitchell and Burrell-( 1 3 4 ) in a n attempt to
understand s p e c i e s relationships through t h e isolation and
identification of antigenic constituents of each s t r a i n by
m e a n s of diffusion and precipitation of antigens and antibodies in gels. They concluded that the o r g a n i s m s which
they studied w e r e serologically i n t e r r e l a t e d and could be
separated into serological subgroups, a s follows:
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2 17
1. H. vaginicola and B. anitratum a r e serologically
identical.
2. Moraxella liquefaciens and M. non-liquefaciens a r e
related.
3. Mima polymorpha and Moraxella lwoffi a r e related.
4. Mima polymorpha var. oxydans i s serologically distinct.
An attempt t o define the antigenic interrelationships of
t h e s e o r g a n i s m s by m e a n s of agglutination, gel diffusion,
and immunofluorescence w e r e described to be "discouraging" by Nelson and Shelton (1965).
Clemesha (19 12, 112) described a motile Gram-positive
o r g a n i s m which produced acid a n d - g a s f r o m glucose and
s u c r o s e . A faint t r a c e of acid was sometimes produced
f r o m lactose; however, this sugar usually was notfermented.
The o r g a n i s m produced acetylmethyl carbinol.
Clemesha
at first "thought it was a l m o s t identical with the ordinary
sewage Proteus"; hence he called it Bacillus P.
Numerous s t r a i n s of Bacillus P (B5W) w e r e isolated and
studied by P a r r (1937a, 1937b), Parr and Galbraith'(19-37),
and Stuart e t al. (1945, 1949). The designation "P bacillus"
was changed to B5W by Stuart ",t al. (1949). According to
S t u a r t ' s recorded description of t h e s t r a i n s he studied and
the attributes of t h e 6 S t u a r t s t r a i n s we have studied (vide
Table l ) , it now a p p e a r s m o r e reasonable to conclude that
C l e m e s h a ' s Bacillus P i s not v e r y closely related to B.anitratum.
The r e s p i r a t o r y f l o r a of patients with bronchitis and
a s t h m a was studied by Henriksen under the direction of
Thjjdtta.
Eleven of the 45 alcaligenes s t r a i n s isolated w e r e
distinctly haemolytic.
The 11 haemolytic s t r a i n s w e r e
homogeneous and sharply distinguishable f r o m the other
alcaligenes s t r a i n s encountered and f o r this r e a s o n w e r e
named Alcaligenes hemolysans ( s i c ) Henriksen (1936, 167).
The s p e c i e s is described a s reducing n i t r a t e to nitrite.
Henriksen p r e p a r e d a r e p r e s e n t a t i v e culture of A. haemolys a n s which was deposited by Thj#tta in 1937 a t the L i s t e r
Institute in London (National Collection of Type Cultures)
but the s t r a i n was discarded in 1958.
The original s t r a i n s
of A. haemolysans a r e no longer available f r o m Henriksen
(correspondence dated 1949 and 1966) and probably a r e no
longer extant.
On two occasions in 1949, before the death of a 6 - y e a r old child with meningitis, Oeding isolated f r o m the c e r e b r o spinal fluid a haemolytic (Oeding s t r a i n 1058, RH 10) and a
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nonhaemolytic (RH 11) o r g a n i s m . Oeding identified s t r a i n
RH 10 as Alcaligenes haemolysans and it was subsequently
reidentified by t h e senior author in 1949 a s a haemolytic
s t r a i n of B. anitratum.
Ferguson et al. (1950) reported
B. a n i t r a t u m s t r a i n s w e r e
that approximately 20% of 109 haemolytic on h o r s e blood a g a r . S t r a i n 11 was v e r y similar
to s t r a i n 10 except for differences i n haemolytic activity;
both w e r e recognized by the s e n i o r author as typical s t r a i n s
of B. anitratum. The ability of t h e s e 2 s t r a i n s to produce
oxidative acidity f r o m glucose was not detected when they
w e r e isolated and d e s c r i b e d by Oeding. It s e e m s reasonable
to suspect that s o m e of t h e ,A. haemolysans s t r a i n s isolated
and d e s c r i b e d by Oeding (1946) m a y have been similar to
s t r a i n 10.
Henriksen in 1950 isolated A . haemolysans s t r a i n 434
(RH 168) f r o m a t h r o a t c u l t u r e of an a s t h m a t i c child. S t r a i n
168 was identified in 1951 by the senior author a s a typical
s t r a i n of B. anitratum.
S t r a i n s l o , 11 and 168 a r e compared with the type s t r a i n
of B. anitraturn, vide Table 3. All 3 s t r a i n s a r e B. -a n i t r a turn. One c a n r e a x y conclude f r o m t h e s e e x p e r i e n c e s that
t h e name Alcaligenes haemolysans s i n c e 1949 m a y have unwittingly applied at l e a s t occasionally to s t r a i n s (RH 10 and
168) of B. anitratum.
The k s t o r i e s of t h e following 4 names, which have been
used by s o m e w o r k e r s a s synonyms for B. anitratum, can
now be discussed.
Diplococcus mucosus von Lingelsheim 1906, 392.
N e i s s e r i a m u c o s a (von Lingelsheim) M u r r a y 19 39, 283.
Achromobacter m u c o s u s (von Lingelsheim) Mannheim
and Stenzel 1962, 75.
Herellea vaginicola De Bord 1942, 476.
Von Lingelsheim (1906, 1908) d e s c r i b e d D. mucosus to be
s m a l l Gram-negative a e r o b i c diplococci o r t e t r a d s which
w e r e distinctly encapsulated.
Gelatin was not liquefied.
Cowan (1938) isolated, described, and named two s t r a i n s of
D. mucosus, which reduced n i t r a t e , produced hydrogen
sulfide, and liquefied gelatin. The s t r a i n s isolated by von
Lingelsheim and Cowan apparently a r e no longer available.
M u r r a y (1939) t r a n s f e r r e d D. m u c o s u s to t h e genus Neisseria.
Seeliger (19 53) received f r o m contemporary European
w o r k e r s , s t r a i n s which had been identified a s D, mucosus.
H e c o m p a r e d them with 11 s t r a i n s of B. a n i t r a t u m which
w e r e received f r o m Ferguson.
These strains formed a
-
-
-
-
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Table 3.
BACTERIOLOGY
219
The c h a r a c t e r i s t i c s of t h r e e misidentified
Alcaligenes s t r a i n s compared with t h e type
s t r a i n of Bacterium anitratum (RH 2208).
RH S t r a i n Number
10
11
Strict aerobe
Gram-negative, rod- shaped
Asporogenous
Atrichous
1-Arabino s e
t
t
t
t
t
t
t
t
t
t
Glucose
Glucose
Glucose
Lactose
Lactose
open
sealed
gas
open
sealed
t
-
t
d-Mannose
d-Mannitol
Sucrose
d-Xylo s e
Indole, methyl red,
acetylmethyl carbinol
t
-
t
-
-
t
t
-
-
Citrate, Simmons
Catalase
Indophenol oxida s e
Gelatin
Hydrogen sulfide, Kligler
t
t
t
t
-
-
t
wkt
t
t
-
Nitrate to g a s
Nitrate to n i t r i t e
Zinc dust t e s t on negative
nitrite tests
Urea, Christensen
Haemoly s is, rabbit blood*-
-
168
t
-
-
t
t
t
-
-
2208
t
-
t
t
+2
-
Alcaligenes haemolysans, Oeding s t r a i n 1058
Nonhaemolytic Alcaligenes, Oeding s t r a i n
Alcaligenes haemolysans, Henriksen s t r a i n 434
Positive t e s t .
Negative test.
Haemolysis around d i s c r e t e s u r f a c e colonies on
infusion agar.
Subscripts = The day on which t h e t e s t became positive.
10 11
168
t
*
-
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homogeneous group and Seeliger suggested t h a t the older
name, D. mucosus, should be used f o r B_. anitratum. V6ron
-e t -al. 6 9 5 9 , 1961) and Ashley and Kwantes (1961) vigorously
object to S e e l i g e r ' s conclusion and recommendation. The
F r e n c h w o r k e r s d e s c r i b e d five s t r a i n s of D. mucosus i s o lated f r o m t h e human r e s p i r a t o r y t r a c t . T h e s e s t r a i n s w e r e
facultative, oxidase positive, t r u e d e n i t r i f i e r s which ' f e r mented glucose and hence w e r e v e r y much unlike the type
s t r a i n of B. anitratum.
Vdron e t al. (1959), Ashley and
Kwantes ( 6 6 1 ) , and Thornley (1967) considered D. mucosus
to be quite different f r o m B. a n i t r a t u m and concluded t h e s e
two n a m e s a r e not synonyms.
Stenzel and Mannheim (1963) proposed NCTC 10303 (RH
2211) a s the neotype s t r a i n for Achromobacter mucosus. If
adopted, a logical extension of t h i s proposal is t h a t RH 2211
m u s t s e r v e a s the neotype for D. mucosus. The type s t r a i n
and 119 other s t r a i n s of B. a n i t r a t u m did not f o r m t e t r a d s ,
did not r e d u c e nitrate, aGd failed to produce hydrogen sulf i d e . S t r a i n 2211 is a rod-shaped o r g a n i s m which slowly
liquefies gelatin, hence does not a p p e a r t o be the s a m e a s
the nomenclatural type (description by von LingeIsheim) of
D. mucosus.
It i s a l s o unlike the s t r a i n s d e s c r i b e d by
Cowan (1938) and V6ron e t al. (1959). It does not s e e m t h a t
NCTC 10303 can be recognized a s t h e neotype s t r a i n for D.
mucosus. Seeliger (1964) r e v e r s e d h i s e a r l i e r recomx-nendation and abandoned t h e u s e of D. mucosus, and a l l other
" n a m e s s o far put forward," for 5.anitratum. On t h e b a s i s
of the above observations we conclude t h a t Diplococcus
mucosus and its two synonyms, N e i s s e r i a m u c o s a and
Achromobacter mucosus, should not be applied t o B,. a n i t r a tum.
S t r a i n 2211 i s compared, in Table 2, with the type s t r a i n ,
RH 2208, of?. anitratum.
The two s t r a i n s produced acid
f r o m xylose in O F m e d i u m as did the 11 B. a n i t r a t u m s t r a i n s
isolated by Schaub and which w e r e among t h e 1 2 0 s t r a i n s
included in t h i s study.
Stuart e t al.
- (1949) r e p o r t e d t h a t
100 B5W s t r a i n s produced acid f r o m xylose.
The tabulated
c h a r a c t e r i s t i c s of t h e s e 2 s t r a i n s do not r e v e a l differences
which can reasonably s e r v e t o s e p a r a t e t h e m at t h e s p e c i e s
level; we conclude that RH 221 1 is a s t r a i n of B. a n i t r a t u m .
T h i s conclusion i s supported by t h e observatiojls of Sm6kal
(1967).
It is evident f r o m t h e above h i s t o r i c a l reviewof the name
D. mucosus that t h e r e is a s yet l i t t l e a g r e e m e n t among bact e r i o l o g i s t s on the nature of t h i s species. T h e r e a p p e a r s to
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221
be sufficient disagreement among bacteriologists to conclude
that the name D. mucosus i s not an e a r l i e r synonym for B.
anit ratum.
De Bord's (1942) description of Herellea vaginicola i s
recorded in Table 2 to facilitate a comparison with the characteristics of the type strain of B_. anitratum. De Bord's
strains of _H. vaginicola apparently a r e no longer available
(personal correspondence from De Bord and others). The
generic name her ellea-the only legitimate species in this
genus i s H. vaginicola-was applied by Deacon (1945) to 10
strains wxich did not produce acid f r o m mannitol o r dulcitol
although he recognized that these organisms failed to agree
with the description of H. vaginicola. Numerous workers
have reported that the tagon described in Table 2 does not
produce acid from mannitol o r dulcitol (Schaub and Hauber
1948; Stuart et al. 1949; Ferguson and Roberts 1950; Brooke
1951; Mitchell and Burrell 1964; Henderson 1965; Elston
and Salisbury 1965; and others); hence it fails to compare
favorably with De Bord's recorded description of H. vaginicola. Since the description of H. vaginicola i s siznificantly
different from the type strain of B. anitratum, the former
name should not be considered a senior synonym for the latter.
This i s not a new observation o r conclusion; however,
the evidence presented in Table 2 offers additional support
for the validity of this conclusion (Henriksen 1960b, 1963;
Hugh and Reese 1967).
The name B. anitratum was a validlypublished legitimate
binary combination. The combination became illegitimate
in 1954 when the generic name Bacterium Ehrenberg was
added to the l i s t of nomina generum bacteriorum rejicienda,
Opinion 4 (Revised). This Opinion of the Judicial Commission did not render the specific epithet anitratum illegitimate. Since neither 2. mucosus nor _H. vaginicola a r e senior
synonyms for B,. anitratum, the later name may be the oldest
known binary combination applied to the taxon concerned in
this study.
Nine synonyms for B. anitratum a r e listed chronologitally below:
-
*Bacterium anitratum Schaub and Hauber 1948, 384.
Achromobacter anitratum (sic) (Schaub and Hauber)
Bourlat 195 3, 43.
Acinetobacter anitratum (sic) (Schaub and Hauber)
Brisou and PrCvot 1954, 727.
Cytophaga anitrata (Schaub and Hauber) Lautrop 1961, 107.
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::Achromobacter conjunctivae Mannheim and Stenzel 19 62,
77.
*Achromobacter haemolyticus subsp. glucidolytica ( s i c )
Mannheim and Stenzel 1962,78.
Achromobacter haemolyticus subsp. haemolyticus
Stenzel and Mannheim 1963, 198.
Moraxella a n i t r a t a (Schaub and Hauber) Lwoff 1964, 483.
Mima a n i t r a t u m (si;) (Schaub and Hauber) Mitchell-and
B u r r e l l 1964,908.
Micrococcus c e r i f i c a n s Finnerty, Hawtrey and Kallio
1962, 169.
T h r e e of the above names a r e r e p r e s e n t e d by type s t r a i n s ,
hence a r e designated with a s t e r i s k s ; they a r e listed in Table
1 and compared i n Table 2.
Achromobacter a n i t r a t u s was a s c r i b e d to Brisou (19 53,
814) by Bourlat (1953, 26) and Buchanan e-t al.
- (1966, l l ) , but
this binary combination does not appear in the 1953 r e p o r t
by Brisou. This combination was u s e d by Bourlat (1953, 43)
and it a p p e a r s t o be the oldest legitimate available name for
B. anitratum.
The justification for assigning B. a n i t r a t u m
to t h e genus Achromobacter i s somewhat difficuit t o a s s e s s
objectively a t this t i m e since t h e r e is reasonable doubt of
the existence of s t r a i n s which can be identified a s A. -liquefaciens, t h e type s p e c i e s of the genus. Recent investigators
have been unable t o find a peritrichous o r g a n i s m that conf o r m s i n a l l r e s p e c t s to the description of t h e basionym
Bacillus liquefaciens Eisenberg and which can s e r v e a s a
neotype (Tulecke
51. 1965, C i t a r e l l a and Colwell 1966,
Thornley 1967). The d e g r e e of relationship between B. anit r a t u m and A. liquefaciens h a s not been made m a n i f e s b y
simultaneous l a b o r a t o r y comparison of s t r a i n s of t h e s e s p e cies.
B. a n i t r a t u m was t r a n s f e r r e d t o t h e genus Acinetobacter
Brisou and Pr6vot (1954,727) and Brisou (1957, 401) designated Acinetobacter a n i t r a t u m ( s i c ) a s t h e type s p e c i e s of
the genus. T h i s nomenclatural type i s based on a s p e c i e s
name that is not validly published (International Code of
Nomenclature of Bacteria, Rule 12). T h e r e i s no r e f e r e n c e
t o a description in t h e r e p o r t by Brisou and PrCvot (1954,
7 27) which differentiates Acinetobacter a n i t r a t u m f r o m the
remaining s p e c i e s and 4 v a r i e t i e s . The name of a s p e c i e s
i s validly published only when i t s publication is accompanied
by a description of t h e s p e c i e s o r by citation of a previously
and effectively published description (International Code of
Nomenclature of Bacteria, Rule 14a). PrCvot ( 196 1, 158) m a y
have recognized this enigma and hence designated Acineto-
zt
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223
bacter stenohalis (Zobell and Upham) Brisou and Pr6vot
1954, 727, a s t h e type s p e c i e s of t h e genus Acinetobacter.
It i s apparent that a genus cannot p o s s e s s 2 type species.
The b a s i s f o r including ,B. a n i t r a t u m in a genus with a type
specieswhich f a i l s to grow in f r e s h w a t e r media and is found
in association with m a r i n e mud and m a r i n e phytoplankton i s
not apparent.
B. a n i t r a t u m and s e v e r a l other s p e c i e s p o s s e s s a kind
of gliding motility (Piechaud 1963; Halvorsen 1963), and
Lautrop (1961) on this b a s i s t r a n s f e r r e d ,B. a n i t r a t u m to t h e
genus Cytophaga.
S t r a i n s of nonflagellated Pseudomonas
aeruginosa, nonflagellated P. fluorescens, Moraxellae, and
Flavobacteria a l s o possess-the kind of twitching s u r f a c e
translocation observed in B. anitratum, and Lautrop (1965)
concluded that B. anitraturn w m n o t y t o p h a g a .
The type .&in(NCTC
10304, RH 2212) of Achromobacter
conjunctivae was designated by Stenzel and Mannheim (19 63).
This s t r a i n i s compared with the type s t r a i n (RH 2208) of
-B. a n i t r a t u m i n Table 2. These 2 type s t r a i n s a r e alike in
a t l e a s t 20 positive and 30 negative c h a r a c t e r s ; they a r e r e markably s i m i l a r . A. conjunctivae should be included in
the species _B. a n i t r a - b .
F o u r c h a r a c t e r s which may d i s tinguish t h e s e 2 s t r a i n s a r e r e c o r d e d in Table 2. These
c h a r a c t e r differences a r e i n t e r p r e t e d t o be infrasubspecific.
-B. a n i t r a t u m is h e r e i n c i r c u m s c r i b e d to accommodate t h i s
biotype.
The name Achromobacter haemolyticus subsp. glucidolytica i s recorded a s a synonym f o r Achromobacter haemolyticus subsp. haemolyticus in a n attempt to bring the name
to conformity with the International Code of Nomenclature.
The type s t r a i n for this subspecies (NCTC 10305, RH 2213)
i s compared in Table 2 with t h e type s t r a i n of B. anitratum.
S t r a i n 2213 is r e m a r k a b l y s i m i l a r to the type-strain of B.
a n i t r a t u m and t h e r e f o r e A. haemolyticus haemolyticus should
be included i n the s p e c i e s B. anitratum. The l a t t e r should
be c i r c u m s c r i b e d to accom-modate o r g a n i s m s with t h i s deg r e e of difference.
The type s p e c i e s of Moraxella i s M. lacunata. It fails to
grow o r responds poorly 'on blood agar, r e q u i r e s a s c i t i c
fluid o r s e r u m f o r growth, produces indophenol oxidase and
f a i l s to f e r m e n t carbohydrate. Piechaud (1963) observed a
gliding type of motility i n z . lacunata and cautioned it was
p r e m a t u r e to u s e this newly observed morphological prop e r t y a s a taxonomic tool. Henriksen (1952, 1960a, 1960b)
and Henriksen and Bbvre (1967) reviewed the l i t e r a t u r e and
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concluded t h e r e is little evidence to include B. a n i t r a t u m in
the genus Moraxella but Lwoff (19 63) proposed the combination M. anitrata.
The type s p e c i e s of the genus Mima De Bord 1942 i s M.
polymorpha, the type s p e c i e s by monotypy.
It is a validly
published legitimate name.
T h e r e is only one legitimate
s p e c i e s in t h i s genus. Mitchell and B u r r e l l (1964,908) t r a n s f e r r e d B. a n i t r a t u m t o t h i s genus a s Mima anitrata. They
considered the older name, H e r e l l e a vaginicola, a synonym
for t h i s new combination; Mima polymorpha v a r . oxidans
was included in Moraxella non-liquefaciens.
S e v e r a l s t r a i n s of alkane-oxidizing Gram-negative m i c r o cocci w e r e isolated f r o m soil, described, and named Micrococcus c e r i f i c a n s Finnerty et al. (1962). T h r e e of t h e s e
original s t r a i n s , vide Table 1, w e r e found t o be v e r y s i m i l a r
to the type s t r a i n of B. anitratum.
T h e s e s t r a i n s could not
be differentiated f r o m t h e type s t r a i n of B. a n i t r a t u m by the
c r i t e r i a used in t h i s study. They a r e t h e r e f o r e considered
tvPical s t r a i n s of B. a n i t r a t u m .
Ach romobacterconjunctivae , Achromobac t e r haemolyt icu s
subsp. glucidolyticus, Achromobacter haemolyticus subsp.
haemolyticus, and Micrococcus c e r i f i c a n s a r e junior synonyms for B. anitratum.
The Su?;committee on Pseudomonas and Related Organi s m s of the International Committee On Nomenclature of
Bacteria suggested the genus Pseudomonas be limited to
s p e c i e s with deoxyribonucleic acid containing 57 to 7 0 m o l e s
percent guanine and cytosine (Shewan and Haynes 1967).
This guanine and cytosine r a n g e would s e r v e to exclude B.
a n i t r a t u m f r o m t h e genus Pseudomonas a s defined by th;
Subcommittee.
-
-
.I
ACKNOWLEDGMENTS
The authors a r e grateful to those who generously provided
c u l t u r e s to make t h i s study possible.
This work was supported in p a r t by r e s e a r c h g r a n t AI07172 f r o m the National Institutes of Health, Department of
Health, Education, and Welfare, U. S. Public Health Service,
U. S. A.
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225
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--
.
-
ADDENDUM
After this m a n u s c r i p t was submitted for publication t h e
name Lingelsheimia a n i t r a t a was proposed by Seeliger ,etal.
1968 (Internatl. J. Syst. Bacteriol. 18:21-32) who stated
t h a t ,B. a n i t r a t u m was placed in the genus Achromobacter
by Brisou and Morichau 1952 (Ann. Inst. P a s t . 82:640-643).
The n a m e Achromobacter a n i t r a t u s does not a p E a r in this
report.
Baumann e-t al.
- 1968 ( J. Bacteriol. 9 5 : 1520- 1541) contend
that B. a n i t r a t u m ATCC 19606 should be accepted a s a synon y m a n d biochemical type of Micrococcus calco-aceticus
ATCC 23055 and that Acinetobacter calco-aceticus i s the
type s p e c i e s of the genus Acinetobacter. Prgvot (1961) designated Acinetobacter stenohalis a s the type species of t h i s
genus.
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