INTERNATIONAL BULLETIN O F BACTERIOLOGICAL
NOMENCLATURE AND TAXONOMY
A p r i l 15, 1964
pp. 69-84
Vol. 14, N o . 2
THE PROPOSED NEOTYPE STRAINS O F PSEUDOMONAS
AERUGINOSA (SCHROETER 1872) MIGULA 1900
Request f o r a n Opinion
Rudolph Hugh' and E i n a r Leifson
The G e o r g e Washington University
School of Medicine, Department of Microbiology
Washington, D. C.
and
Loyola University, School of Medicine a n d G r a d u a t e School
Department of Microbiology, Chicago, Ittinois
R e s e a r c h C o l l a b o r a t o r s , A m e r i c a n T y p e C u l t u r e Collection
SUMMARY. A n h i s t o r i c a l r e v i e w o f t h e n a m e
Pseudomonas aeruginosa (Schroeter) Migula
is presented.
The morphological and physiologic a 1 c h a r a c t e ri st i c s , i n c luding e le ct r on
of
a
micrographs
and a photomicrograph
stained preparation, a r e presented in Lysenko's proposed neotype strain (ATCC 10145,
IBCAS 2 7 7 , N R R L
NCTC 1 0 3 3 2 , NCIB 8295,
B - 7 7 1 , C C E B 4 8 1 , R H 8 1 5 ) of P s e u d o m o n a s
aeruginosa.
This strain is compared with
several other proposed neotype strains. The
authors support the proposal by Lysenko and
request an Opinion on this proposal by the
Judicial
C o m m i s s i o n of
the
International
Committee on Bacteriological Nomenclature.
The work of the s e n i o r author was supported i n p a r t by
r e s e a r c h g r a n t E-3186 f r o m the National I n s t i t u t e s of
Health, Department of Health, Education a n d We lfare, U .
S. Public HeaLth S e r v i c e , to the A m e r i c a n Type C u l t u r e
Collection.
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
P a g e 70
INTERNATIONAL B U LLE TIN
INTRODUCTION
The b l u e - g r e e n p u s o r g a n i s m w a s f i r s t n a m e d B a c t e r i u m
a e r u g i n o s u m ( S c h r o e t e r 1872,126).
The d e s c r i p t i o n given
by S c h r o e t e r i s a s follows:
P. 109: "Die k l e i n e n O r g a n i s m e n , welche am haufigsten i n
i h r e n bewegten F o r m e n a l s B a k t e r i e n , i n i h r e n unbewegten
a l s B a k t e r i d i e n b e z e i c h n e t w e r d e n , s t i m m e n d a r i n tiberein,
d a s s s i e stickstoffhaltige, o r g a n i s c h e Stoffe ( P r o t o p l a s m a m a s s e n ) mit g r o s s e r E n e r g i e z u r eigenen E r n g h r u n g v e r b r a u c h e n , und d a b e i s p e c i f i s c h e Stoffe m a n n i g f a c h e r A r t
bilden.
"Von d i e s e n P r o d u k t e n f a l l e n eine Reihe von P i g m e n t e n
m i t a m m e i s t e n i n die Augen.
E s sind Lebhafte F z r b u n g e n
der verschiedensten Art, die wir aus farblosen ProtoplasmaGebilden e n t s t e h e n s e h e n , a l l e i n b e g l e i t e t von mas s e n h a f t e r
Entwicklung s o l c h e r k l e i n e r O r g a n i s m e n , d i e w i r d e m n a c h
a l s atleinige U r s a c h e d e r Farbenbildung ansehen miissen.
"Eine R e i h e von K u l t u r e n , d i e g r b s s t e n t e i l s i n d e n Wint e r m o n a t e n 1868 b i s 1870 im pflanzenphysiologischen I n s t i tute zu B r e s l a u v o r g e n o m m e n w u r d e n , h a t t e n den Zweck,
Beobachtungen a b e r d e r a r t i g e Pigmentbildungen zu s a m m e l n .
Im folgenden s o l l e n k u r z die e r h a l t e n e n R e s u l t a t e n a c h d e r
Reihe d e r F a r b e n z u s a m m e n g e s t e l l t w e r d e n .
''
P. 1 2 2 : "Grtin. Auf a u s g e l e g t e n Stiicken g e k o c h t e r K a r t o f f e l n sah i c h e i n i g e m a l e i n e f l e c k w e i s e GrUnfZrbung e i n t r e t e n .
Die F a r b e w a r e i n d u n k e l e s Saftgriin, s i e begann am Rande
d e r Scheiben, b r e i t e t e s i c h e x c e n t r i s c h a u s und d r a n g e t w a s
i n d i e N a h r s u b s t a n z ein. E i n e S c h l e i m a u f l a g e r u n g w a r nicht
vorhanden, B a k t e r i e n v e r m o c h t e i c h nicht aufzufinden. So
s e h r i c h a u c h geneigt bin, d i e s e Pigment-Bildung b e i A b w e s e n h e i t j e d e r a n d e r e n U r s a c h e , auf B a k t e r i e n z u r u c k z u f u h r e n , s o kann i c h s i e doch n u r a l s zweifelhaft i n d e n K r e i s
die s e r B e t r a c h t u n g e n z i e h e n .
"Eine g e n a u e r bekannte P r o d u k t i o n von griinem F a r b stoff
d u r c h B a k t e r i e n findet i n d e m sogenannten griinen E i t e r
s t a t t . Nach d e n von A n d e r e n d a r u b e r v o r g e n o m m e n e n Unt e r suchungen, w i r d e r d u r c h bewegte B a k t e r i e n gebildet.
I c h habe m e h r f a c h griinen E i t e r a u f t r e t e n s e h e n , a b e r nicht
i n g r o s s e r A u s b r e i t u n g und u n t e r U m s t z n d e n , d i e mir e i n e
m i k r o s k o p i s c h e und c h e m i s c h e P r u f u n g nicht m 6 g l i c h m a c h t e n . Die F a r b e w a r i n d i e s e n F a l l e n i m m e r g l e i c h m z s s i g
...
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
P a g e 71
BACTERIOLOGICAL NOMENCLATURE
AND TAXONOMY
s p a n g r u n , e t w a s n a c h blau u b e r g e h e n d . Auffallig w a r d i e ,
s c h o n a n d e r e r s e i t s h e r v o r g e h o b e n e T a t s a c h e , dass das
P i g m e n t n i c h t sowohl a n d e m E i t e r haftet, d e r d i e B a k t e r i e n
e n t h a l t , s o n d e r n i n die F a d e n d e r aufgelegten C h a r p i e und
K o m p r e s s e n e i n z i e h t und d i e s e i n w e i t e r Ausdehnung f a r b t
E s geht d a r a u s h e r v o r , d a s s a u c h h i e r nicht d i e B a k t e r i e n
T r g g e r d e s F a r b s t o f f e s sind, s o n d e r n dass e r von d i e s e n i n
d i e umgebende F l u s s i g k e i t a u s g e s c h i e d e n w i r d . . . ' I
,
" E s i s t v i e l l e i c h t nicht u n b e r e c h t i g t b e i j e d e r
P. 126:
b e s t i m m t e n P i g m e n t b i ldung einen s p e cifi s c h v e r s c hiedenen
O r g a n i s m u s anzunehmen, und d e m g e m a s s n e b e n e i n e m
B a c t e r i d i u m prodigiosum (Ehrbg. ) auch ein B a c t e r i d i u m
aurantiacum,
Luteum, c y a n e u m , v i o l a c e u m , b r u n n e u m ,
neben B a c t e r i u m synxanthus und B,. syncyanus ( E h r b g . ) e i n
B,. a e u r u g i n o s u m a u f z u s t e l l e n . I '
.
. .
.
S c h r o e t e r (1 872) r e p o r t e d t h e p r o d u c t i o n of a w a t e r s o l u b l e , b l u e - g r e e n p i g m e n t on s l i c e d boiled potato a n d i n
p u s . S c h r o e t e r acknowledged the w o r k of o t h e r s (without
documentation) who ob s e r v e d t h a t m o t i l e b a c t e r i a p r o d u c e
g r e e n pigment a n d g r e e n pus. It is c l e a r that S c h r o e t e r
(1 872) n e i t h e r m i c r o s c o p i c a l l y o b s e r v e d n o r i s o l a t e d the
o r g a n i s m which he n a m e d B a c t e r i u m a e r u g i n o s u m . I t m i g h t
be s a i d t h a t the a p p l i c a t i o n of the n a m e B_. a e r u p i n o s u m
S c h r o e t e r i s u n c e r t a i n . However i n n u m e r a b Le s u b s e q u e n t
i n v e s t i g a t o r s have i s o l a t e d , c u l t u r e d , a n d o b s e r v e d t h e
o r g a n i s m r e s p o n s i b l e f o r t h e p r o d u c t i o n of b l u e - g r e e n p u s
a n d have c h a r a c t e r i z e d t h e c a u s a t i v e o r g a n i s m a n d the
w a t e r - s o l u b l e p i g m e n t (pyocyanine) it p r o d u c e s . C o n t e m p o r a r y b a c t e r i o l o g i s t s r e c o g n i z e only one s p e c i e s ( P s e u d o m o n a s a e r u g i n o s a ) which p r o d u c e s pyocyanine. Doubt conc e r n i n g the a p p l i c a t i o n of t h e n a m e B a c t e r i u m a e r u g i n o s u m
S c h r o e t e r i s t h e r e f o r e quickly d i s p e l l e d b e c a u s e no o t h e r
o r g a n i s m h a s been i m p l i c a t e d i n t h e p r o d u c t i o n of pyocyanine o r b l u e - g r e e n pus. I t is not c l e a r why C o h n ( 1 8 7 2 , 1 5 7 )
r e f e r r e d to B a c t e r i u m a e r u g i n o s u m S c h r o e t e r a s B a c t e r i d L m aerugineum (sic).
The n a m e P s e u d o m o n a s w a s p r o p o s e d by Migula (1894,
237) f o r a genus which i n c l u d e d a single s p e c i e s , _P. v i o l a c e a , which o r d i n a r i l y would b e c o m e t h e type s p e c i e s of the
genus (by monotypy). T h e r e i s no citation t o a p r e v i o u s l y
d e s c r i b e d s p e c i e s with t h i s n a m e n o r i s t h e r e a n y clue a s
t o which of t h e nine o r m o r e p r e v i o u s l y d e s c r i b e d s p e c i e s
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
P a g e 72
INTERNATIONAL
BULLETIN
bearing the specific epithet violacea the n a m e _P. violacea
Migula i s to apply. The E d i t o r i a l B o a r d (Buchanan, B r e e d ,
and Cowan) of the Judicial Commission of the International
Committee on B a c t e r i o l o g i c a l Nomenclature (1951) concluded that under Rule 14 of the International Bacteriologic a l Code of Nomenclature t h e s p e c i e s name Pseudomonas
violacea Migula 1894 was not validly published, and under
Rule 24 the name i s illegitimate.
Migula (1895 p. 29) d e s c r i b e d the genus P s e u d o m o n a s i n
g r e a t e r detail and listed 11 previously n a m e d s p e c i e s , the
f i r s t of which was Pseudomonas pyocyanea ( G e s s a r d ) Migula
1895 with t h e synonym B a c i l l u s pyocyaneus G e s s a r d .
Migula (1900 p . 884) published the new combination
Pseudomonas aeruginosa (Schroeter 1872) Migula 1900 with
the f ollowiag synonyms : B a c t e r i u m a erugino s u m Schr o e t e r
1872, B a c i l l u s a e r u g i n o s u s S c h r o e t e r 1886, B a c i l l u s pyocyaneus G e s s a r d 1882b. Migula i n c o r r e c t l y a s c r i b e d the
binary combination B a c i l l u s a e r u g i n o s u s t o S c h r o e t e r 1886
p. 157. B a c i l l u s aeruginosus (Schroeter 1872) should be
attributed to T r e v i s a n 1885 p. 101.
The J u d i c i a l Commission of the International Committee
on Bacteriological Nomenclature (1 952 Opinion 5) c o n s e r v e d
the g e n e r i c n a m e Pseudomonas Migula 1894 and designated
Pseudomonas aeruginosa(Schroeter 1872) Migula 1900 a s the
type s p e c i e s with t h e following synonyms: B a c t e r i u m a e r u ginosum S c h r o e t e r 1872, B a c i l l u s pyocyaneus Ge s s a r d 1882a,
Pseudomonas pyocyanea ( G e s s a r d ) Migula 1895 p. 29. The
citations f o r the l a s t two synonyms a r e i n c o r r e c t and should
be Pseudomonas pyocyanea (Zopf) Migula 1895.
Zopf (1884 p. 83) a s c r i b e s Micrococcus pyocyaneus to
G e s s a r d 1882b. Migula (1895 p. 29) and Matzuschita (1902
p . 1 3 2 ) a s c r i b e B a c i l l u s pyocyaneus to G e s s a r d . Mace' (1889
p.495), Migula (1900 p . 884), and Mace' (1901 p. 852) a s c r i b e
B a c i l l u s pyocyaneus t o G e s s a r d 1882b. B e r g e y g t 51. (1923
p. 123) and B r e e d e_t 4, (1957 p. 99) a s c r i b e B a c i l l u s ~
o
cyaneus to G e s s a r d 1882a. The Judicial Commission of the
Internationa 1 Committee on B a c t e riologica 1 Nomenclature
(1952, Opinion 5 p. 121), a s c r i b e Bacillus pyocyaneus to
G e s s a r d 1882a. Examination of the G e s s a r d 1882a and 1882b
r e f e r e n c e s c l e a r l y show that the b i n a r y combination Micrococcus pyocyaneus a n d B a c i l l u s pyocyaneus w e r e not u s e d
by G e s s a r d . The t e r m Bac-pyocyaneus a, was u s e d by G e s s a r d (1890), and E r n s t (1887) u s e d the n a m e B a c i l l u s pyocyaneus (Zopf) FlUgge 1886.
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
-
P a g e 73
BACTERIOLOGICAL NOMENCLATURE
AND TAXONOMY
Seven s y n o n y m s of P s e u d o m o n a s a e r u g i n o s a ( S c h r o e t e r
1892) Migula 1900, 884 a r e :
B a c t e r i u m a e r u g i n o s u m S c h r o e t e r 1872, 126
B a c t e r i d i u m a e r u g i n e u m ( s i c ) Cohn 1872, 157
M i c r o c o c c u s pyocyaneus Zopf 1884, 83
B a c i l l u s a e r u g i n o s u s ( S c h r o e t e r ) T r e v i s a n 1885, 101
B a c i l l u s pyocyaneus (Zopf) FlUgge 1886, 286
P s e u d o m o n a s pyocyanea (Zopf) Migula 1895, 29
B a c t e r i u m pyocyaneum (Zopf) L e h m a n n a n d Neumann
1896, 267
I n a s m u c h a s S c h r o e t e r 1872 did not i s o l a t e B a c t e r i u m
a e r u g i n o s u m , the e x i s t e n c e of a type s t r a i n i s i m p o s s i b l e .
T h e a l t e r n a t i v e i s to e s t a b l i s h , a c c o r d i n g to p r e s c r i b e d
p r o c e d u r e , a neotype s t r a i n . I n t h i s i n s t a n c e the neotype
s t r a i n m u s t : 1 ) p r o d u c e pyocyanin, a n d 2 ) be a m o t i l e r o d
( B a c t e r i u m ) . S c h r o e t e r (1872) o b s e r v e d t h e w a t e r - s o l u b l e
a n d blue-green pigment of B a c t e r i u m a e r u g i n o s u m on boiled
,P. a e r u g i n o s a i s t h e only s p e c i e s of
potato a n d i n p u s .
b a c t e r i a known to p r o d u c e w a t e r - s o l u b l e b l u e - g r e e n pigm e n t (pyocyanine). Subsequent w o r k h a s d e m o n s t r a t e d t h a t
P. a e r u g i n o s a i s G r a m - n e g a t i v e , r o d - s h a p e d , a n d m o t i l e
by m e a n s of a p o l a r f l a g e l l u m , T h e r e i s g e n e r a l a g r e e m e n t
t h a t ,P. a e r u g i n o s a h a s t h e s e c h a r a c t e r i s t i c s .
During t h e p a s t 5 y e a r s , s e v e r a l s t r a i n s have b e e n s u g g e s t e d i n t h e l i t e r a t u r e a s p r o p o s e d neotype s t r a i n s o r t e m p o r a r y "working type s t r a i n s " of _P. a e r u g i n o s a .
Sneath
a n d Cowan(1958) d e s i g n a t e d NCTC 6750 a t e m p o r a r y w o r k ing type s t r a i n f o r _P. a e r u g i n o s a pending t h e e s t a b l i s h m e n t
of a neotype s t r a i n by the I n t e r n a t i o n a l C o m m i t t e e on B a c teriological Nomenclature.
T h i s s t r a i n p r o b a b l y n e e d not
b e c o n s i d e r e d s e r i o u s l y a s a s u i t a b l e neotype s t r a i n s i n c e
Sneath a n d Cowan (1958) a l s o s u g g e s t e d
i t is atrichous.
NCTC 2000 a s a r e f e r e n c e s t r a i n f o r ,P. a e r u g i n o s a . L y s e n ko (1961) p r o p o s e d s t r a i n CCEB 481 a s t h e neotype s t r a i n .
On the b a s i s of m e d i a n o r g a n i s m h y p o t h e s i s s t u d i e s , L i s t o n
e t ~ 1 (1963)
.
r e c o m m e n d e d e i t h e r A T C C 14209 o r A T C C
14216 as a s u i t a b l e neotype s t r a i n . O t h e r s ( p e r s o n a l c o m m u n i c a t i o n s ) a r e of the opinion that old s t o c k s t r a i n s m i g h t
be m o r e suitable than r e c e n t i s o l a t e s a s candidates f o r a
neotype s t r a i n a n d s u g g e s t t h a t the following ,P. a e r u g i n o s a
s t r a i n s , a l l of which w e r e i s o l a t e d b e f o r e 1925, be con-
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
P a g e 74
INTERNATIONAL
BULLETIN
sidered: ATCC 97, 256, 260 and 262. T h e r e a r e no r u l e s
o r recommendations in the International Bacteriological
Code of Nomenclature to indicate that a n old s t r a i n i s to be
p r e f e r r e d over a r e c e n t isolate a s a neotype s t r a i n . The
singular advantage of a n older s t r a i n is t h a t i t might be
m o r e extensively documented than a r e c e n t i s o l a t e . In the
c a s e a t hand t h e r e a p p e a r s to be no r e a l advantage to the
selection of an older s t r a i n a s the neotype; i t i s our opinion
that ability to produce pyocyanine i s a m o r e important consideration.
This point is d i s c u s s e d below.
However the
only r e q u i r e m e n t of a neotype i s that i t a g r e e with the diagnosis given by the original d e s c r i b e r .
The purpose of this p a p e r 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 the above eight Pseudomonas aeruginosa s t r a i n s
(ATCC 97, 256, 260, 262, 10145, 14149, 14209 and 14216)
and to suggest which one of the eight candidate s t r a i n s would
m o s t adequately s e r v e a s the neotype s t r a i n .
MATERIALS AND METHODS
Tabte 1 gives some of the corresponding s t r a i n n u m b e r s
of the 8 ,P. aeruginosa s t r a i n s studied.
The h i s t o r i e s of
these s t r a i n s a r e given i n TabLe 2 . Kavanagh and Woodruff
Table 1. Corresponding n u m b e r s of the 8 Pseudomonas a e r u g i n o s a
strains.
ATCC
97
256
260
262 10145 14149 14209
RH
145
146
147
148
815
1091
1448
NCTC
10332
2000
NRRL
B-247 B-249 B-249 B-250 B-771
van den Ende
NCIB
8295
CCEB
481
LB CAS
277
Drake
44
Key:
ATCC
NCTC
NRRL
NCLB
CCEB
IB CAS
14216
1455
A m e r i c a n Type Culture Collection, Washington, D. C .
National Collection of Type C u l t u r e s , London
N o r t h e r n Regional R e s e a r c h L a b o r a t o r y , P e o r i a
National Co LLe ction of Industria 1 B a c t e r i a , Ab e r d e e n
Culture Collection of Entomogenous B a c t e r i a , P r a g u e
Institute of Biology, Czechoslovak Academy of Sciences,
Prague
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
D
Table 2.
T h e h i s t o r i e s of 8 P s e u d o m o n a s a e r u g i n o s a s t r a i n s .
ATCC
Received
strain
at A T C C
number as strain
Date
deposited
a t ATCC
Deposited
Remarks
by
1925
Walter Reed
A r m y Medical
Center
Jordan strain.
1925
E.O. Jordan
I s o l a t e d (1898) by LeCount f r o m g a n g r e n o u s
t i s s u e of a p a t i e n t with l e u k e m i a .
260 LeCount IV
1925
E . 0. J o r d a n
I s o l a t e d by LeCount f r o m a n a m p u t a t i o n s t u m p
of a p a t i e n t with l e u k e m i a .
262
1925
E.O. Jordan
Gessard strain
10145
1946
F. Kavanagh
Used i n antibiotic sensitivity studies.
1947 J . B a c t e r i o l . %:761-766.
1961
S . T . Cowan
NCTC 2000, 1958 J. Gen. M i c r o b i o l . g : 5 5 1 - 6 5
D r a k e s t r a i n 4 4 . 1960 J. B a c t e r i o l . @:420-21
97
256
14149
151
Eyre I
14209
44
1961
C.H. Drake
14216
D
1961
R . M . Wood
v a n d e n Ende s t r a i n D of h u m a n o r i g i n .
1952 J. Hyg. 5 0 ~ 4 0 5 - 4 1 4
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
Page 76
INTERNATIONAL
BULLETIN
w e r e unable to provide information concerning the date and
s o u r c e of isolation of _P. aerupinosa ATCC s t r a i n 10145.
The p r o c e d u r e s u s e d to c h a r a c t e r i z e the 8 s t r a i n s of _P.
aeruginosa have been d e s c r i b e d by Hugh and Ryschenkow
(1961). Blood a g a r b a s e (Difco 0045) which contains beef
h e a r t infusion was used to detect pyocyanine production.
RESULTS
Each of the 8 Pseudomonas aeruginosa s t r a i n s studied
have the same 26 positive c h a r a c t e r i s t i c s and the s a m e 23
negative c h a r a c t e r i s t i c s enumerated i n Table 3 . Some of
t h e s e 8 s t r a i n s a r e positive, while o t h e r s a r e negative i n
r e s p e c t t o the 8 c h a r a c t e r i s t i c s e n u m e r a t e d i n Table 4.
ATCC s t r a i n 10145 i s d e s c r i b e d below:
Morphology: Gram-negative, asporogenous, rod-shaped;
c e l l s s t r a i g h t with s o m a s i z e 0 . 5 x 1.5-2.5 p.
Motile by
m e a n s of predominantly one polar flagellum having a m e a n
wavelength of 1 . 6 p, ( F i g s . 1, 2, 3 i n t h e p l a t e ) , Two flagella
may occasionally be found a t one pole of some c e l l s . F o r malin t r e a t m e n t does not significantly change the f lage Llar
shape. Capsules a r e not evident on the media employed in
the study.
Physiology: This s t r i c t a e r o b e produces a dense turbidity i n n e u t r a l peptone broth i n 18-24 hours a t t e m p e r a t u r e s
of 22, 37 a n d 41 "C, but f a i l s to grow at 25 to 37°C in b r a i n
h e a r t infusion b r o t h a t pH 4 . 5 . I t i s neither halophilic n o r
halophobic. T h e r e is good growth i n broth containing potass i u m cyanide.
An anaerogenic a c i d reaction i s produced i n unsealed
tubes of Oxidative F e r m e n t a t i v e (OF) B a s a l Medium (Difco
0668, Hugh and Leifson 1953) containing the following c a r bon compounds: a r a b i n o s e , dextrose, ethanol 370,f r u c t o s e
g a lacto s e , g lyce r o 1, mannitol, mannos e, m e libio s e , d - r i b
o s e , t r e h a l o s e , and xylose. The acid reaction i s apparent
in 1-3 days except with t r e h a l o s e which may r e q u i r e 1 0 days
of incubation before a c i d a p p e a r s . An acid reaction i s p r o duced i n unsealed tubes of O F dextrose medium containing
0 . 0 0 1 m o l a r iodoacetic acid sodium salt. An a c i d reaction
i s not produced i n OF d e x t r o s e medium s e a l e d with p e t r o latum. C i t r a t e (Simmons and Christensen), c a t a l a s e , indopheno 1 oxida s e , ge Latin, 2 - ke tog Luconat e , m a Lonate, nit r a t e
-
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
Page 77
BACTERIOLOGICAL NOMENCLATURE
A N D TAXONOMY
Similar C h a r a c t e r i s t i c s of 8 Pseudomonas aeruginosa s t r a i n s .
Table 3 .
97
260
t
t
t
t
t
t
f
+
+
+r
-
+2
+2
-
t
-
+
-
_
-
-
+
+
+
+
+
+2
t
t
+b
+
Gram-negative rod-shaped
t
Motility
t
F l a g e l l a r wavelength 1.5-2 p t
P o l a r monotrichous
t
Growth a t 22.37 and41'C
Adonitol
-
+
Arabino s e
Ce llobiose
Dextrose open
Dextrose sealed
Dextrose g a s
Dextrose with iodoacetic
acid
Du lcito I
Ethanol, 3%
Galactose
G ly c e r o 1
Inositol
Inulin
Maltose
Manno s e
Me le zito s e
Melibiose (Difco)
Raffinose
d-Ribose
Salicin
Sorbitol
Sucrose
Xylose
Indole
Methyl r e d
Acetylmethyt carbinol
Citrate -Simmons
Citrate -Christensen
Catala s e
Indophenol oxidase
Gelatin-charcoal
Hydrogen sulfide-Kligler
2-ketogluconate
Lysine decarboxylaseC a r lquis t
Ma lona t e
Nitrate to gas
N i t r a t e to nitrite
Phenylalanine deamina s e
Urea-Christensen
Lysine-Mbller
Arginine Mbller
Ornithine -Mbller
Growth a t pH 4 . 5
Spores
-
ATCC Strain Number
262
10145
14149 14209
256
-
+
-
-
-
t
t
t
-
+
-
+
-
t
+
+
_
-
-
-
t
t
+
f,
f
i
t 2
+
-
-
-
-
-
-
-
t
t
+
$2
-
+2
-
+2
+2
+2
t
-
-
-
-
-
2
t
t
t
t
+
+
+ , +
-
f
t
t
+3
-
+3
+t
+
t
i
+
t
t
-
-
-
f2
-
t
t2
+
-
-
,
+
t
+
+
+
t
+
+
+
t
+
+
+
t
t
+
+
+
t
t
+
+
-
t
+
t
t
+
+
+
-
t
+
+
t
t
t
+
-
+2
-
+
t
f
+
t
14216
t
+
+
i
t
+2
+2
+
+
+
+
+
+
t
+
+
t
t
+
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
P a g e 78
INTERNATIONAL BULLETIN
EXPLANATION O F T H E P I A T E
F i g u r e 1.
P s e u d o m o n a s a e r u g i n o s a A T C C 10145 showing
the p o l a r m o n o t r i c h o u s flagellation of t h e o r g a n i s m f r o m b r o t h c u l t u r e without f o r m a l i n fixation.
T h e s o m a of t h i s s t r a i n g e n e r a l l y s t a i n s v e r y
faintly. L e i f s o n f l a g e l l a s t a i n , X2, 000.
F i g u r e s 2 a n d 3 . P s e u d o m o n a s a e r u g i n o s a A T C C 10145
showing a r o d f r o m b r o t h c u l t u r e with a p o l a r
m o n o t r i c h o u s f Lagellum. E l e c t r o n m i c r o g r a p h ,
p a l l a d i u m shadowed, X22, 000.
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
Page 79
BACTERIOLOGICAL NOMENCLATURE
AND TAXONOMY
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
Page 80
INTERNATIONAL
Table 4.
BULLETIN
Dissimilar c h a r a c t e r i s t i c s of 8 Pseudomonas aeruginosa s t r a i n s .
ATCC Strain Number
97
Fructose
Lactose
Mannito 1
Rhamnose
T r ehalo s e
Growth in KCN broth
Water soluble pigment
Chloroform so tub Le
wk+,
t
t7
256
wktz
-
wkt
-
-
260
262
-
-
t
-
t4
-
-
-
-
10145
14149
14209
t
t
t
t
tll
+zo
+in
+?
-
+7
t
+I0
t
tttt
tttt
t
t
t
t
+a
t
14216
t
t
wk t
Key:
positive t e s t on the f i r s t day; subscripts indicate the day t e s t became positive
t
negative test
wk weak
tttt strongly positive
-
reduction to n i t r i t e and nitrogen gas, u r e a (Christensen),
and arginine (Mbller) t e s t s a r e positive.
Pyocyanine is
visibly produced in infusion agar (Difco 0045) and i s readily
e x t r a c t e d with c h l o r o f o r m .
An a c i d reaction i s not produced i n tubes of unsealed O F
b a s a l medium with t h e following carbon compounds: adonitot, c e llobiose, dulcitol, inositol, inulin, lactose, maltose,
melezitose, raffinose, Rhamnose, salicin, sorbitot, and
s u c r o s e . The indole, methyl red, acetylmethyt carbinol,
hydrogen sulfide (Kligler, a f t e r 2 days’ incubation), Lysine
decarboxylase (ninhydrin), phenylalanine deaminase, lysine
and ornithine (MblLer) t e s t s a r e negative.
DISCUSSION
It is a p p a r e n t that the 8 candidate s t r a i n s of Pseudomonas aeruginosa a r e v e r y homogeneous i n r e s p e c t to the
49 c h a r a c t e r i s t i c s enumerated a n d compared i n Table 3 .
They a r e alike i n 2 6 positive and 2 3 negative c h a r a c t e r i s t i c s . The s t r a i n differences r e c o r d e d i n Table 4 a r e not
i n t e r p r e t e d to be of profound significance in taxonomy except f o r pigment production, which m a y r e q u i r e some d i s cussion as it r e l a t e s to the selection of the neotype s t r a i n .
Some s t r a i n s of ,P. aerupinosa readily produce large
quantities of pyocyanine i n a l m o s t e v e r y medium capable of
supporting growth.
Other s t r a i n s a r e apyocyanogenic even
when grown i n many s p e c i a l media devised to induce pigment
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
P a g e 81
BACTERIOLOGICAL NOMENCLATURE
A N D TAXONOMY
production ( G e s s a r d , Burton, Paton, S e l l e r s , King, e t c . ) .
Between t h e s e two e x t r e m e s a r e s t r a i n s which a p p e a r to be
apyocyanogenic except when grown on one of the s p e c i a l
m e d i a devised t o induce pigment production. E a c h pseudomonadologist a p p e a r s to p r e f e r h i s own f a v o r i t e pyocyanineinducing medium since t h e r e i s little evidence that one m e dium i s u n i v e r s a l l y s u p e r i o r i n inducing pyocyanine production in a l l s t r a i n s . It should be o b s e r v e d that _P. a e r u g i n o s a
s t r a i n s c a n be identified even though pyocyanine i s not p r o duced.
P. a e r u g i n o s a s t r a i n s 97, 256, 260, 262, 14149, 14209,
and-1421 6 did not produce visible water-soluble pigment on
the infusion a g a r medium employed i n this study ( s e e Table
4). However, a v e r y s m a l l quantity of pyocyanine w a s ext r a c t a b l e with chloroform f r o m t h e infusion a g a r m e d i u m
on which s t r a i n 14216 had grown f o r 7 d a y s , Pyocyanine
could not be e x t r a c t e d f r o m t h e infusion a g a r medium on
which s t r a i n s 97, 256, 260, 262, 14149, and 14209 had grown
f o r 7 days. Haynes (1951) r e p o r t e d that s t r a i n s 97, 257, 260
and 262 w e r e apyocyanogenic in G e s s a r d ' s a n d B u r t o n ' s
media, however these s t r a i n s produce pyocyanine i n S e l l e r s
medium. E a c h of the 8 ,P. a e r u g i n o s a . s t r a i n s produce pyocyanine in a t least one of t h e s p e c i a l pigment-inducing m e d i a .
None of t h e 8 s t r a i n s a r e d r a m a t i c a l l y p r o l i f i c pigment p r o d u c e r s . However, of t h e s e 8 s t r a i n s ATCC 10145 i s unusual
i n that i t r e a d i l y p r o d u c e s visible pigment i n infusion a g a r
medium, the u s u a l routine medium employed i n the clinical
diagnostic laboratory. This blue pigment i s r e a d i l y e x t r a c t e d with chloroform.
The quantitative equivalence of a l l c h a r a c t e r i s t i c s to
e s t a b l i s h phenons i n n u m e r i c a l taxonomy has c r e a t e d the
p r o b l e m of evaluating the significance of the v a r i o u s phenon
levels in t e r m s of the c l a s s i c a l l y established s p e c i e s . L i s ton g t a_L. (1963) published observations on the o v e r - a l l
s i m i l a r i t y , e x p r e s s e d i n S values, of 3 3 s t r a i n s of ,P. a e r u ginosa a s c o m p a r e d to a calculated hypothetical m e d i a n o r ganism, and Lockhart g t a l . (1963) emphasized the concept
of monothetic c o r e p r o p e r t i e s a s the c h a r a c t e r i s t i c f e a t u r e
of a s p e c i e s , t h e s e p r o p e r t i e s being n o r m a l l y distributed
among s t r a i n s . The 17 m i n i m a l - c o r e p r o p e r t i e s d e s c r i b e d
by Liston e t ~ 1 (1963)
.
r e p r e s e n t t h e m i n i m u m calculated
hypothetical o r g a n i s m f o r ,P. a e r u g i n o s a .
,P. aeruginosa
s t r a i n s 14209 and 14216 w e r e r e p o r t e d by L i s t o n g t 51. (1963)
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
Page 82
INTERNATIONAL
BULLETIN
to show 9770 s i m i l a r i t y t o t h e i r concept of the calculated
median o r g a n i s m w h e r e a s s t r a i n 10145 h a s 82% s i m i l a r i t y .
The specific differences in the c h a r a c t e r i s t i c s of s t r a i n
14216 and Lysenko's proposed neotype s t r a i n 10145 a r e not
readily a p p a r e n t f r o m t h e data p r e s e n t e d in t h e r e p o r t by
Liston c t at. (1963).
Undoubtedly m a n y s t r a i n s of _P. aeruginosa c a n m e e t the
r e q u i r e m e n t s of a neotype s t r a i n . E a c h of the 8 s t r a i n s
studied p o s s e s s e s the monothetic-core p r o p e r t i e s of ,P.
a e r u g i n o s a ; however, e a c h would not be equally useful a s a
neotype s t r a i n of ,P. a e r u g i n o s a .
A s a r e s u l t of t h i s c o m p a r a t i v e study of the 8 candidate
s t r a i n s and p a r t i c u l a r l y because of pigment production,
ATCC s t r a i n 10145 is judged to be the m o s t suitable to r e commend f o r the neotype s t r a i n of Pseudomonas aeruginosa.
Lysenko (1961) proposed Pseudomonas a e r u g i n o s a ATCC
10145 a s the neotype. The a u t h o r s support t h i s p r o p o s a l a n d
h e r e b y request a n Opinion on this p r o p o s a l by the Judicial
Commission of the International Committee on B a c t e r i o logical Nomenclature.
ACKNOWLEDGMENTS
The a u t h o r s a r e indebted to E r w i n F. L e s s e l , C u r a t o r of
B a c t e r i a , A m e r i c a n Type Culture Collection, f o r g e n e r o u s
advice, encouragement, and a s s i s t a n c e with t h e m a n u s c r i p t .
The e l e c t r o n m i c r o g r a p h s w e r e p r e p a r e d by the kind effort
of R i c h a r d C. F o w l e r , P r o f e s s o r of Medicine, The George
Washington University Cancer Clinic.
REFERENCES
B e r g e y , D.H., F . C . Harris,on, R.S. B r e e d , B.W. H a m m e r
and F. Huntoon. 1923. B e r g e y ' s Manual of Determinative Bacteriology. The Williams and Wilkins Co., B a l t i m o r e . 1 s t ed. 1-442.
B r e e d , R . S . , E.G.D. M u r r a y , N . R . Smith e t at. 1957.
B e r g e y ' s Manual of Determinative Bacteriology. The
Williams a n d WiLkins Co., B a l t i m o r e . 7th ed. 1-1094.
Cohn, F. 1872. Untersuchungen Uber B a c t e r i e n . B e i t r z g e
zur Biologie d e r Pflanzen, 1 . Band, 2. Heft, 127-224.
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
P a g e 83
BACTERIOLOGICAL NOMENCLATU R E
AND TAXONOMY
E d i t o r i a l B o a r d . 1951. S t a t u s of P s e u d o m o n a s MiguLa 1894
a s a g e n e r i c n a m e , a n d of P s e u d o m o n a s a e r u g i n o s a
( S c h r o e t e r ) Migula 1900 a s type s p e c i e s . I n t e r n a t l . B u l l .
B a c t . Nomencl. and Taxon. 1:41-42.
E r n s t , P. 1887. U e b e r e i n e n neuen B a c i t l u s d e s b l a u e n
E i t e r s ( B a c . p y o c y a n e u s (3), eine S p i e l a r t d e s B a c .
p y o c a n . d e r A u t o r e n . Z e i t s c h r . Hyg. 2:369-381.
FLiigge, C . 1886. Die M i k r o o r g a n i s m e n . F. C. W. Vogel,
L e i p z i g . 2.Aufl. 1-692.
G e s s a r d , M. C. 1882a. S u r l e s c o l o r a t i o n s b l e u e e t v e r t e
d e s tinges 2 p a n s e m e n t s . Compt. Rend. A c a d . S c i . ,
Paris %:536-538.
1882b. De la Pyocyanine e t de Son M i c r o b e , T h k s e
inaugual d e la F a c u k e ' d e me'de'cine d e Paris. N u m b e r
248, 1-68.
. 1890. S u r Le M i c r o b e pyocyanique. A n n a l e s de
L'Ins titut P a s t e u r 4: 8 8- 10 2.
H a y n e s , W. C. 1951. P s e u d o m o n a s a e r u g i n o s a - i t s
chara c t e r i z a t i o n a n d identification. J o u r . Gen. Microbiol.
5~939-950.
Hugh, R . a n d E . L e i f s o n . 1953. The taxonomic s i g n i f i c a n c e
of f e r m e n t a t i v e v e r s u s oxidative m e t a b o l i s m of c a r b o h y d r a t e s by v a r i o u s G r a m - n e g a t i v e b a c t e r i a . J o u r .
B a c t . 66:24-26.
a n d E. Ryschenkow. 1961. P s e u d o m o n a s m a l t o p h i l i a ,
a n A l c a l i g e n e s -Like s p e c i e s . J o u r . G e n . Microbiol.
26~123-132.
Hoff, J . C . a n d C.H. D r a k e . 1960. Susceptibility of
P s e udo m o na s po lyc o l o r to P s eudomon a s a e r ugi no s a
bacteriophages. Jour. Bact. g:420-421.
J u d i c i a l C o m m i s s i o n , Opinion No, 5 . 1952. C o n s e r v a t i o n
of t h e g e n e r i c n a m e P s e u d o m o n a s MiguLa 1894 a n d d e s i g nation of P s e u d o m o n a s a e r u g i n o s a ( S c h r o e t e r ) Migula
1900 a s type s p e c i e s . I n t e r n a t t . B u l l . B a c t . N o m e n c l .
and Taxon. 2 ~ 1 2 1 - 1 2 2 .
Kavanagh, F. 1947. A c t i v i t i e s of twenty-two a n t i b a c t e r i a l
s u b s t a n c e s a g a i n s t nine s p e c i e s of b a c t e r i a , J o u r . B a c t .
54~761-766.
L e h m a n n , K . B . a n d R . Neumann. 1896. A t l a s und G r u n d r i s s d e r B a k t e r i o t o g i e und L e h r b u c h d e r s p e c i e t l e n
b a k t e r i o l o g i s c h e n Diagnostik. J. F. L e h m a n n , Milnchen.
1. A u f l . T e i l 11: Text, 1-448.
.
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51
P a g e 84
INTERNATIONAL BULLETIN
Liston, J . , W . Wiebe and R . R . Colwell. 1963. Quantitative
a p p r o a c h to the study of b a c t e r i a l s p e c i e s . J o u r . B a c t .
85~1061-1070.
Lockhart, W.R. a n d P . A . H a r t m a n . 1963. F o r m a t i o n of
monothetic g r o u p s i n quantitative b a c t e r i a l taxonomy.
J o u r . B a c t . 85:68-77.
Lysenko, 0. 1961. Pseudomonas-An a t t e m p t a t a g e n e r a l
classification. J o u r . Gen. Microbiol. 25 :3 7 9-408,
Mace', E . 1889. T r a i t 6 pratique de bactkriologie. J . - B .
B a i l l i g r e e t f i l s , Paris. 1 ed. 1-711.
1901. T r a i t 6 p r a t i q u e de bact6riologie. J.-B.
Baillie're and f i t s , P a r i s . 4th ed. 1-1196.
Matzuschita, T 190 2 B a c t e r i o logi sche Diagno stik.
G. F i s c h e r , J e n a . 1-690.
Migula, W . 1894. Ueber e i n n e u e s S y s t e m d e r B a k t e r i e n .
A r b e i t e n a u s d e m Bakteriologischen Institut d e r Technichen Hochschule zu K a r l s r u h e . l.Band, 235-238.
1895. Schizomycetes ( B a c t e r i a , B a c t e r i e n ) , in:
A . E n g l e r a n d K. P r a n t l , Die Naturlichen Pflanzenfamilien, 1, Teil, Abt. l a , 1-44.
1900. S y s t e m d e r B a k t e r i e n . G. F i s c h e r , J e n a .
2.Band, 1-1068.
S c h r o e t e r , J. 1872. U e b e r einige d u r c h B a c t e r i e n gebildete
P i g m e n t e . in: F . Cohn, B e i t r a g e z u r Biologie d e r
Pflanzen, L B a n d , 2.Heft, 109-126.
1886. Schizomycetes (Naegli 1857). i n : F . Cohn,
Kryptogamen-Flora von Schlesien, 3,Band, 2,Abt. 136174.
Sneath, P . H . A . and S. T. Cowan. 1958. An e l e c t r o taxonomic s u r v e y of b a c t e r i a . J o u r . Gen. Microbiol.
19~551-565.
T r e v i s a n , V . 1885. C a r a t t e r i di alcuni nuovi g e n e r i d i
B a t t e r i a c e e . Atti della A c c a d e m i a Fisio-MedicoStatistica i n Milano, S e r i e 4, 2:92-106.
van den Ende, M. 1952. O b s e r v a t i o n s on the antigenic
s t r u c t u r e of P s e u d o m o n a s a e r u g i n o s a . J o u r . Hyg. 50:
40 5 -41 4.
Zopf, W . 1884, Die Spaltpilze. E . Trewendt, B r e s l a u .
2.Aufl., 1-98.
.
.
.
.
.
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Fri, 16 Jun 2017 17:07:51