INTERNATIONAL BULLETIN O F BACTERIOLOGICAL
NOMENCLATURE AND TAXONOMY
A p r i l 15, 1964
pp. 87-101
Vol. 14, No. 2
THE PROPOSED CONSERVATION O F THE GENERIC
NAME VIBRIO PACINI 1854 AND DESIGNATION O F THE
NEOTYPE STRAIN O F VLBRIO CHOLERAE PACINI 1854
R e q u e s t f o r a n Opinion
Rudolph Hugh
The G e o r g e Washington U n i v e r s i t y School of Medicine
Department of Microbiology, Washington, D. C.
R e s e a r c h C o l l a b o r a t o r , A m e r i c a n Type C u l t u r e Collection
SUMMARY. V i b r i o M u t t e r h a s n o t b e e n u s e d i n
t h e L i t e r a t u r e of p r o t o z o o l o g y f o r o v e r 1 0 0
y e a r s . A r t i c l e 2 3 of t h e I n t e r n a t i o n a l Z o o l o g i c a l C o d e of N o m e n c l a t u r e s a y s " a n a m e t h a t
has remained unused as a senior synonym in
the primary zoological literature for more
than 50 y e a r s is t o be c o n s i d e r e d a f o r g o t t e n
name (nomen oblitum)."
For a little more
than 100 years Vibrio has become established
as a commonly accepted generic name in bacteriology. It is recommended f r o m the standp o i n t of b a c t e r i o l o g y t h a t V i b r i o M u t t e r 1 7 7 3
b e r e g a r d e d a s t h e n a m e of a g e n u s i n z o o l ogy and in conformity with the International
C o d e of Z o o l o g i c a l N o m e n c l a t u r e b e r e c o g nized as a nomen oblitum a n d without standing in bacteriological nomenclature.
It is
further recommended that Vibrio Pacini 1854
b e p l a c e d i n t h e l i s t of c o n s e r v e d g e n e r i c
n a m e s (nomina generum conservanda) with the
type species Vibrio cholerae Pacini 1854 and
t h a t t h e s p e c i f i c e p i t h e t c h o l e r a e in t h e b i n a r y
combination Vibrio cholerae Pacini 1854 also
be conserved.
It is also proposed that the
N a t i o n a l C o l l e c t i o n of T y p e C u l t u r e s ( N C T C )
strain 8021 (American Type Culture Collection (ATCC) strain 14035) be recognized as
t h e n e o t y p e s t r a i n of V i b r i o c h o l e r a e P a c i n i
1854.
The morphological and physiological
c h a r a c t e r i s t i c s , i n c Luding a p h o t o m i c r o g r a p h
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of a s t a i n e d p r e p a r a t i o n s h o w i n g m o n o t r i c h o u s
cells each with a polar flagellum, a r e pres e n t e d of t h e p r o p o s e d n e o t y p e s t r a i n .
- - - - - - - - -
MUller (1773, 1786) u s e d binomina i n the n o m e n c l a t u r e
and c l a s s i f i c a t i o n of m i c r o s c o p i c o r g a n i s m s fotlowing the
publication by Linnaeus of the 1758 edition of S y s t e m a Natur a e . Many of M u t t e r ' s g e n e r a a n d s p e c i e s a r e recognized
a n d i n u s e today. Some of h i s o r g a n i s m s a r e known t o b e
protozoa, s o m e a r e now placed with nematodes, s o m e of h i s
g e n e r a a n d s p e c i e s a r e recognized a s a l g a e . The d e s c r i p tions and i l l u s t r a t i o n s of h i s s p e c i e s m a k e i t highlyprobabLe
that s o m e w e r e b a c t e r i a . MUller's (1773, 39) genus V i b r i o
i s of p a r t i c u l a r i m p o r t a n c e f r o m the standpoint of b a c t e r i ology. In t h i s genus, p l a c e d i n h i s Infusoria c r a s s i u s c u l a '
(Mutter 1773, 23), he included 15 s p e c i e s with n a m e s and
d e s c r i p t i o n s . Some of the s p e c i e s n a m e d by MUller (1773,
1786) w e r e l a t e r p l a c e d i n other g e n e r a , first of protozoa,
serpens,
then of b a c t e r i a . F o r example V i b r i o rugula,
V . undula, and 41. s p i r i l l u m w e r e placed by E h r e n b e r g (1830,
3 8 ) a n d W i n t e r (1884, 63) i n the genus S p i r i l l u m .
Leidy (1849) examined the i n t e s t i n a l contents of the m i l l i pede ( J u l u s m a r g i n a t u s ) , t h e toad (Bufo a m e r i c a n u s ) , a n d
the contents of the c a e c u m of the r a t , a n d a p p e a r s to be the
f i r s t to conclude that V i b r i o lineola MCIlLer 1773, 39 a n d
S p i r i l l u m undula (MUller) E h r e n b e r g 1830, 38, found t h e r e i n ,
a r e p l a n t s r a t h e r than a n i m a l s . Buchanan (1925, 522) p r o p o s e d that the s p e c i e s whose n a m e a n d d e s c r i p t i o n c a m e
f i r s t in Mutter's (1773, 39) listing, Vibrio Lineola, b e d e s i g nated a s the type s p e c i e s . Although d e s c r i b e d a s a n infuso r i a n , Mtlller's d e s c r i p t i o n m a k e s probable that t h i s o r g a n i s m w a s a schizomycete.
The g e n e r i c n a m e Vibrio Milller 1773 h a s been a r b i t r a r i l y u s e d i n s e v e r a l s e n s e s by different w r i t e r s ; Vuillemin
(1913) reviewed the p r o b l e m a n d suggested that the g e n e r i c
n a m e should be s u p p r e s s e d to avoid f u r t h e r confusion. If
Vibrio i s r e g a r d e d a s a genus of protozoa, the f a c t that the
v.
C r a s s i u s c u l a i s the diminutive of the c o m p a r a t i v e of the
Latin adjective c r a s s u s (thick, d e n s e , not flowing, o r a m o e boid) meaning "infusoria that a r e somewhat m o r e thick o r
solid. "
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type s p e c i e s cannot be identified, and the f u r t h e r f a c t that
p r e s e n t - d a y p r o t o z o o l o g i s t s do not a c c e p t o r u s e the n a m e
V i b r i o Mutter 1773 would m e a n that the a c c e p t a n c e of the
n a m e i n zoology i s subject to the r u l e s of the I n t e r n a t i o n a l
Code of Zoological N o m e n c l a t u r e . A r t i c l e 23 of the zoolo g i c a l code s t a t e s that "a n a m e that h a s r e m a i n e d u n u s e d
a s a s e n i o r synonym i n t h e p r i m a r y zoological l i t e r a t u r e
f o r m o r e than 50 y e a r s i s t o b e c o n s i d e r e d a f o r g o t t e n n a m e
(nomen oblitum).
A s u r v e y of " p r i m a r y zoological l i t e r a t u r e " b a s e d upon
u s a g e d u r i n g the p a s t 50 y e a r s (1914-1964) has r e v e a l e d n o
u s e of V i b r i o a s a g e n e r i c n a m e i n zoology. While i t w a s
u s e d a s a g e n e r i c n a m e somewhat m o r e than 100 y e a r s ago
i n naming c e r t a i n o r g a n i s m s a s p r o t o z o a now p l a c e d with
t h e b a c t e r i a (Naegali 1857, Cohn 1872), i t s e e m s c l e a r t h a t
a s a g e n e r i c n a m e i t h a s not been r e c o g n i z e d i n zoology f o r
m o r e than 100 y e a r s . In a c c o r d a n c e with A r t i c l e 23 of the
zoological code t h e r e i s no r e a s o n t o r e g a r d t h e g e n e r i c
n a m e V i b r i o Mutter 1773 a s a s e n i o r synonym i n p r i m a r y
zoological l i t e r a t u r e and t h e r e should b e no o b s t a c l e u n d e r
the b a c t e r i o l o g i c a l code t o i t s u s e a s a g e n e r i c n a m e i n
b a c t e r i o l o g y . A r e c o m m e n d a t i o n m a d e i n c o n f o r m i t y with
A r t i c l e 23 of the I n t e r n a t i o n a l Code of Zoological Nomenc l a t u r e w i l l be f o r m u l a t e d a n d published.
The question t o b e c o n s i d e r e d next i s t h e validity a n d
legitimacy of the recognition of Vibrio a s a g e n e r i c n a m e i n
b a c t e r i o l o g y with the goal of fixing i t s s t a t u s .
E h r e n b e r g ( 1 8 3 8 ) u s e d the i n f u s o r i a n g e n e r i c n a m e V i b r i o
f o r o r g a n i s m s which w e r e l a t e r r e c o g n i z e d a s Schizomyc e t e s . V i b r i o synxanthus E h r e n b e r g (1 840, 202) a n d V i b r i o
syncyanus E h r e n b e r g (1840, 202) have been t r a n s f e r r e d f r o m
t h i s protozoon genus t o the b a c t e r i a l genus P s e u d o m o n a s
Migula 1894 ( c o n s e r v e d 1952, Opinion 5). F u c h s (1841,190)
xanthogenus;
n a m e d a n d d e s c r i b e d v i b r i o cyanogenus a n d
a c c o r d i n g t o T r e v i s a n (1885,104) a n d S c h r o e t e r (1886,157)
t h e s e n a m e s a r e j u n i o r synonyms of the o r g a n i s m s c u r r e n t l y
c l a s s i f i e d a s P s e u d o m o n a s syncyanea ( E h r e n b e r g ) Migula
1895,29 and P. synxantha ( E h r e n b e r g ) B r e e d 1948, 700 r e spectively a n d hence i l l e g i t i m a t e . Most of the s p e c i e s of
b a c t e r i a included i n t h e genus V i b r i o by w r i t e r s b e f o r e 1854
a r e now definitely p l a c e d i n o t h e r g e n e r a o r cannot be identified.
P a c i n i (1 8 5 4 ) a p p e a r s t o be the f i r s t t o d e s c r i b e a n o r g a n -
v.
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i s m which i s r e c o g n i z e d today a s a s p e c i e s of b a c t e r i a c u r r e n t l y c l a s s i f i e d i n the genus V i b r i o . The etiologic agent
of A s i a t i c c h o l e r a w a s f i r s t d e s c r i b e d a n d n a m e d v i b r i o
c h o l e r a ( s i c ) by P a c i n i 1 8 5 4 , 4 1 1 . He w a s the f i r s t to u s e
t h e g e n e r i c n a m e V i b r i o with a l e g i t i m a t e s p e c i f i c epithet,
c h o l e r a e , f o r t h e b a c t e r i u m which i s t h e c a u s a l a g e n t of
Asiatic c h o l e r a . Filippo Pacini, f o r many y e a r s P r o f e s s o r
of Pathology a t t h e Royal I n s t i t u t e of S u p e r i o r Studies i n
F l o r e n c e , s t u d i e d A s i a t i c c h o l e r a a f t e r i t s 1854 a p p e a r a n c e
i n T u s c a n y a n d r e m a i n e d i n t e r e s t e d i n t h i s d i s e a s e f o r the
r e m a i n d e r of h i s life. B i a n c h i (1885) published, a f t e r the
d e a t h of P a c i n i , the unedited l a b o r a t o r y n o t e s a n d i l l u s t r a tions which P a c i n i m a d e while e x a m i n i n g c a d a v e r s of c h o l e r a
v i c t i m s i n F l o r e n c e . P a c i n i s a w the c u r v e d etiologic a g e n t
of A s i a t i c c h o l e r a i n t r e m e n d o u s n u m b e r s i n denuded i n t e s t i n a l e p i t h e l i u m a n d i n t e s t i n a l c o n t e n t s of v i c t i m s . T h e
pathological c h a n g e s i n t h e i n t e s t i n e a n d the o r g a n i s m i t s e l f
w e r e d e s c r i b e d i n s u c h a m a n n e r t h a t t h e y m a y be r e c o g nized.
P a c i n i ' s c o n t e m p o r a r y Count V i t t o r e T r e v i s a n , a
s y s t e m a t i c b a c t e r i o l o g i s t , (1 884, 373; 1884, 427) had no
doubts that P a c i n i had d e s c r i b e d a n d n a m e d t h e etiologic
a g e n t of A s i a t i c c h o l e r a . The g e n e r i c n a m e V i b r i o should
t h e r e f o r e be a s c r i b e d to P a c i n i 1854 a n d should b e c o n s e r v e d
a g a i n s t V i b r i o Milller 1773 a n d a l l o t h e r zoological n a m e s
f o r the taxon to avoid f u r t h e r c h a n g e s i n the n o m e n c l a t u r e
of the g e n u s . V i b r i o c h o l e r a e P a c i n i 1854, the type s p e c i e s
by monotypy, should a l s o be c o n s e r v e d .
Koch (1884) w a s the f i r s t to i s o l a t e t h e etiologic agent of
Asiatic cholera.
He gave a m o r e d e t a i l e d d e s c r i p t i o n of
t h e o r g a n i s m a n d u s e d t h e v e r n a c u l a r n a m e KommabaciLlus
i n s t e a d of the e a r t i e r s c i e n t i f i c n a m e V i b r i o c h o l e r a e P a c i n i
1854. Koch went to Egypt without p r e v i o u s knowledge of
c h o l e r a o r of i t s i n f e c t i o u s n a t u r e . He d i d not know that
P a c i n i had d e s c r i b e d a n d n a m e d V i b r i o c h o l e r a e 30 y e a r s
e a r l i e r . T r e v i s a n w a s convinced of the i d e n t i t y of V i b r i o
c h o l e r a e P a c i n i a n d K o c h ' s K o m m a b a c i l l u s which was r e c o v e r e d f r o m E g y p t i a n s with A s i a t i c c h o l e r a . T r e v i s a n was
a w a r e that P a c i n i (1854) h a d p l a c e d a b a c t e r i u m i n t h e zoolo g i c a l g e n u s V i b r i o MUller 1 7 7 3 a n d he t r a n s f e r r e d V i b r i o
c h o l e r a e P a c i n i t o t h e b a c t e r i a l g e n u s B a c i l l u s a s B_. chole r a e ( P a c i n i ) T r e v i s a n 1884, 374. The g e n e r i c n a m e P a c i n i a
w a s p r o p o s e d by T r e v i s a n (1885, 8 3 ) w i t h t h e A s i a t i c c h o l e r a
o r g a n i s m a s the f i r s t s p e c i e s n a m e d .
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Zopf t r a n s f e r r e d B a c i l l u s cho l e r a e -as ia t i c a e T r e v i s a n
1884 to t h e genus S p i r i l l u m a s S. c h o l e r a e - a s i a t i c a e ( T r e v i s a n ) Zopf 1 8 8 5 , 6 9 .
Schroeter proposed the g e n e r i c n a m e
M i c r o s p i r a a n d r e n a m e d the etiologic agent of A s i a t i c c h o l e r a , M i c r o s p i r a c o m m a S c h r o e t e r 1886,168.
Blanchard
(1906, 1 ) a p p e a r s to b e the f i r s t t o u s e t h e s p e c i f i c e p i t h e t
c o m m a with the genus V i b r i o .
S e v e r a l a u t h o r s ( B l a n c h a r d 1 9 0 6 , l ; C a s t e l l a n i a n d Chalm e r s 1919, 962; K r a s i l ' n i k o v 1949, 530) d e s c r i b e d t h e e t i o logic a g e n t of A s i a t i c c h o l e r a a s the first s p e c i e s i n the
genus V i b r i o ; o t h e r s a c t u a l l y d e s i g n a t e d t h i s s c h i z o m y c e t e
a s the type s p e c i e s of the genus V i b r i o (Buchanan 1918, 1 7 8 ;
.
77; E n d e r l e i n 1925,255; P r i b r a m 1933,
B e r g e y Et ~ 1 1923,
4 9 ; Wilson a n d M i l e s 1946, 514; Hauduroy 1953, 648; Wilson
a n d M i l e s 1955,604; B r e e d e t 51. 1957,229; Sneath a n d c o w a n
1958, 554; Haupt 1 9 6 4 , 4 8 ) .
T h e following n a m e s ( s y n o n y m s ) h a v e b e e n u s e d f o r :
Vibrio cholerae P a c i n i 1854,411.
B a c i l l u s c h o l e r a e ( P a c i n i ) T r e v i s a n 1884, 374.
B a c i l l u s c h o l e r a e - a s i a t i c a e T r e v i s a n 1884; a c c o r d i n g to
T r e v i s a n 1885, 84.
B a c i l l u s v i r g u l a ( s i c ) T r e v i s a n 1884, 376.
P a c i n i a c h o l e r a e - a s i a t i c a e ( T r e v i s a n ) T r e v i s a n 1885, 84.
S p i r i l l u m c h o l e r a e - a s i a t i c a e ( T r e v i s a n ) Zopf 1885, 69.
Microspora c o m m a Schroeter 1886,168.
S p i r i l l u m c h o l e r a e ( P a c i n i ) Mace' 1889, 606.
V i b r i o c h o l e r a e - a s i a t i c a e ( T r e v i s a n ) P f e i f f e r 1896, 527.
Liquidovibrio c h o l e r a e ( P a c i n i ) O r l a - J e n s e n 1909, 333.
V i b r i o c o m m a ( S c h r o e t e r ) B Lanchard 1906, 1 .
-B a c i l l u s c o m m a ( S c h r o e t e r ) Holland 1920, 218.
The National Collection of Type C u l t u r e s s t r a i n 8021
(ATCC 14035, RH 1094) i s h e r e i n p r o p o s e d a s t h e neotype
s t r a i n of V i b r i o c h o l e r a e P a c i n i 1854 (vide Sneath a n d Cowan
1958; Hugh a n d Ryschenkow 1961). T h i s s t r a i n c o n f o r m s to
the d e s c r i p t i o n of t h e etiologic agent of A s i a t i c c h o l e r a a s
given by P a c i n i (1854) a n d Koch (1884).
MATERPALS AND METHODS
The p r o p o s e d neotype s t r a i n w a s d e p o s i t e d i n the NCTC
by t h e S t a n d a r d s L a b o r a t o r y , C e n t r a l P u b l i c H e a l t h L a b o r a -
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t o r y , Colindale, London, N.W.9, i n S e p t e m b e r 1949 a s
Vibrio c h o l e r a e a s i a t i c a e , G a r d n e r a n d v e n k a t r a m a n 0 group
I, Ogawa 0 f o r m (Kauffmann 1950). A descendant of t h i s
s t r a i n w a s deposited i n ATCC i n 1960. The d a t e a n d s o u r c e
of isolation of this s t r a i n w a s not obtainable f r o m t h e NCTC.
Leifson's (1951, 1960) technique w a s employed to s t a i n f l a ge 1La
The effects of the o r g a n i s m on s u g a r s a n d alcohols w a s
d e t e r m i n e d i n p u r p l e b r o t h b a s e (Difco 0227) containing 170
(weight p e r volume, w/v) of e a c h of the 2 2 c a r b o n c o m pounds. Aesculin w a s added t o p u r p l e b r o t h b a s e i n a 0.5vo
(w/v) concentration. Modified Kovacs' r e a g e n t (Gadebusch
a n d G a b r i e l 1956) w a s u s e d to d e t e c t indole i n 48-hour 170
(w/v) tryptone b r o t h c u l t u r e . Buffered peptone glucose b r o t h
i n 3 m l and 1 m l quantities was inoculated f o r the methyl
r e d (MR) a n d V o g e s - P r o s k a u e r (VP) t e s t s r e s p e c t i v e l y . The
MR b r o t h w a s incubated f o r 5 d a y s , and t h e V P b r o t h f o r
2 days before the t e s t s w e r e p e r f o r m e d a s d e s c r i b e d by
Ewing (1960). C i t r a t e utilization w a s d e t e r m i n e d i n S i m m o n s ' (1926) and C h r i s t e n s e n ' s (1949) m e d i u m a n d f i n a l
observations w e r e r e c o r d e d a f t e r 2 days incubation. Hydrogen peroxide (370,w / v ) w a s dropped on 24-hour nutrient a g a r
slant growth to detect c a t a l a s e production. The indophenol
oxidase t e s t , a s d e s c r i b e d by Ewing and Johnson (1960), w a s
p e r f o r m e d on d i s c r e t e 24-hour colonies on infusion a g a r .
Kohn's (1953) c h a r c o a l gelatin was u s e d to d e t e c t gelatinolytic activity. Kligler's i r o n a g a r w a s u s e d to d e t e c t hydrogen sulfide production, and final o b s e r v a t i o n s w e r e m a d e
a f t e r 2 days of incubation, The ninhydrin t e s t w a s p e r f o r m e d
a c c o r d i n g t o the method of C a r l q u i s t (1956). Malonate utilization w a s d e t e r m i n e d by the u s e of Leifson's (1933) m a l o n a t e b r o t h m o d i f i e d by the addition of 0.1% ( w / v ) y e a s t e x t r a c t
a s d e s c r i b e d by Ewing, Davis a n d R e a v i s (1957). The f i n a l
observation was m a d e a f t e r 7 days incubation.
N i t r a t e reduction was d e t e r m i n e d i n b r o t h a t pH 7 . 1 containing 170(w/v) casitone, 0. 370 (w/v) y e a s t e x t r a c t , a n d 0.270
(w/v) p o t a s s i u m n i t r a t e . The b r o t h was dispensed i n 4 m l
quantities i n 13 x LOO mm t e s t tubes containing D u r h a m v i a l s
f o r the collection of g a s a n d autoclaved a t 1 2 1 ° C f o r 15 min.
C u l t u r e s w e r e examined f o r n i t r a t e reduction to n i t r i t e a f t e r
2 d a y s of incubation by adding the r e a g e n t s d e s c r i b e d by
Wallace and Neave (1927) and T i t t s l e r (1930). The phenylalanine d e a m i n a s e t e s t w a s p e r f o r m e d a f t e r 24 h o u r s of
.
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incubation a s d e s c r i b e d by Ewing, Davis a n d R e a v i s (1957).
Ability t o g r o w i n the p r e s e n c e of p o t a s s i u m cyanide w a s
d e t e r m i n e d by using M b l l e r ' s (1954) m e d i u m a n d f i n a l obs e r v a t i o n s w e r e m a d e a f t e r 2 d a y s of incubation. C h r i s t e n s e n ' s (1946) a g a r w a s u s e d t o d e t e c t u r e a h y d r o l y s i s a n d
f i n a l o b s e r v a t i o n s w e r e r e c o r d e d a f t e r 2 d a y s incubation.
F a l k o w ' s (1958) d e c a r b o x y l a s e m e d i u m containing lysine,
arginine and ornithine w a s used. A control and the 3 amino
aci.d-fortified m e d i a w e r e i n o c u l a t e d a n d l a y e r e d with s t e r i l e , m e l t e d p e t r o l a t u m to a d e p t h of 10-20 mm. F i n a l obs e r v a t i o n s w e r e m a d e a f t e r 7 d a y s of incubation. H e m o l y s i s
of washed s h e e p e r y t h r o c y t e s w a s e s t i m a t e d by t h e t e s t tube
technique using a 24-hour c u l t u r e grown i n p h y s i o l o g i c a l
p a n c r e a t i c d i g e s t of c a s e i n . The r e s u l t s w e r e r e a d a f t e r
incubation a t 3 7 ° C f o r two h o u r s followed by 1 8 h o u r s of
r e f r i g e r a t i o n a t 6" C.
All media w e r e incubated a t 37°C.
The c r i t e r i a f o r i n t e r p r e t a t i o n of p o s i t i v e a n d n e g a t i v e t e s t s w e r e d e s c r i b e d
i n the r e f e r e n c e c i t e d and a m p l i f i e d by Ewing (1960) a n d
E d w a r d s a n d Ewing (1962).
RESULTS
Morphology: G r a m - n e g a t i v e , a s p o r o g e n o u s , r o d - s h a p e d ;
cells straight o r slightly c u r v e d w i t h soma s i z e 0 . 3 to 0 . 6 x
1 t o 5 p . Motile with p r e d o m i n a n t l y one p o l a r f l a g e l l u m
having a m e a n wavelength of 2.4 p ( s e e F i g s . 1, 2).
Cells
with two f l a g e l l a a t one pole o r one f l a g e l l u m a t both p o l e s
a r e r a r e l y encountered.
F o r m a l i n t r e a t m e n t d o e s not s i g nificantly change the f l a g e l l a r s h a p e . C a p s u l e s a r e not e v i dent on t h e m e d i a e m p l o y e d i n t h i s s t u d y ,
Physiology: This o r g a n i s m g r o w s a e r o b i c a l l y i n peptone
b r o t h f r e e of f e r m e n t a b l e c a r b o h y d r a t e a n d f a c u l t a t i v e l y i n
peptone containing d e x t r o s e . I t f a i l s to g r o w on desoxycholate a g a r .
D e x t r o s e i s f e r m e n t e d without g a s p r o d u c t i o n . A c i d i s
p r o d u c e d f r o m d e x t r o s e u n d e r a n a e r o b i c a n d a e r o b i c conditions. Acid is produced f r o m fructose, galactose, lactose,
m a l t o s e , m a n n i t o l (without g a s ) , m a n n o s e , s u c r o s e , a n d
t r e h a l o s e . A t r a n s i e n t weak acidity i s produced in sorbitol
b r o t h . The l a c t o s e a n d t r e h a l o s e r e a c t i o n s a r e d e l a y e d f r o m
2 to 4 d a y s . Indole, c i t r a t e , c a t a l a s e , indophenol o x i d a s e ,
ge Latin, l y s i n e d e c a r b o x y l a s e (ninhydrin), n i t r a t e r e d u c t i o n
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F i g u r e s 1 and 2 . Vibrio cholerae (NCTC 8021) showing the
polar monotrichous f lage Llum c h a r a c t e r i s t i c of the
o r g a n i s m i n broth culture with f o r m a t i n fixation.
Leifson f lage lla s t r a i n , X 2 , 0 0 0 .
to n i t r i t e , lysine and ornithine (Falkow) t e s t s positive. The
methyl r e d indicator i n the MR t e s t becomes orange r a t h e r
than r e d o r yellow. The V P t e s t i s weakly positive.
Acid i s not produced f r o m adonitol, aesculin, a r a b i n o s e ,
ce LLobio se , dulcitol, inositol, inulin, me le eitose, m e libios e,
raffinose, rhamnose, salicin, and xylose Hydrogen sulfide,
malonate, n i t r a t e reduction to nitrogen gas, phenylalanine
deaminase, u r e a , arginine t e s t s a r e negative. The organi s m does not grow i n potassium cyanide broth. Sheep e r y throcytes a r e not hemolysed and the c e l l s a r e agglutinated
by 0 - g r o u p I antibodies.
a
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DISCUSSION
B a c t e r i o p h a g e s t u d i e s ( M u k e r j e e 1961, 1963) have i n d i c a t e d that the etiologic a g e n t of c l a s s i c a l A s i a t i c c h o l e r a i s
g e n e r a l l y s u s c e p t i b l e to a s p e c i f i c b a c t e r i o p h a g e (Group IV)
w h e r e a s EL T o r v i b r i o s a r e l y s o - r e s i s t a n t to t h i s s a m e
b a c t e r i o p h a g e . Stab le nonhemolytic V i b r i o c h o l e r a e s t r a i n s
which a r e i n s e n s i t i v e to G r o u p IV b a c t e r i o p h a g e w e r e i s o lated d u r i n g the 1962 New Guinea e p i d e m i c . Hugh (1962)
r e p o r t e d that 0 - g r o u p I h e m o l y t i c E l T o r v i b r i o s ( V i b r i o
e l t o r E i s l e r 1909, 549) w e r e a biotype (physiological type)
of V i b r i o c h o l e r a e P a c i n i 1854. The r u l e s of n o m e n c l a t u r e
c l e a r l y r e c o m m e n d t h a t biotypes a n d phagotypes (phage
t y p e s ) be r e c o g n i z e d a s i n f r a s u b s p e c i f i c subdivions of a
species
C o m p a r a t i v e t a x o n o m i c s t u d i e s a p p e a r to a g r e e t h a t o r g a n i s m s which f a i l t o p r o d u c e a n a e r o b i c a c i d i t y f r o m d e x t r o s e should be excluded f r o m t h e genus V i b r i o and t h a t the
following p r o v i s i o n a l definition of the genus V i b r i o i s w a r ranted:
G r a m - n e g a t i v e ; a s p o r o g e n o u s s t r a i g h t r o d s o r r o d s with
a single r i g i d c u r v e ; p o l a r f l a g e lla; p r o d u c e a n a e r o b i c a c i d i t y f r o m d e x t r o s e without g a s p r o d u c t i o n . I t i s r e c o g n i z e d
t h a t t h i s i n c o m p l e t e definition m a y include a f e w s p e c i e s of
o t h e r g e n e r a but f u r t h e r s t u d i e s a p p e a r to be n e c e s s a r y bef o r e a g r e e m e n t m a y be r e a c h e d on how t h i s definition m a y
b e e m e n d e d t o include o r exclude o r g a n i s m s ( e . g . A e r o m o n a s f o r m i c a n s ( C r a w f o r d ) P i v n i c k a n d Sabina 1957, A e r o monas c a v i a e ( S c h e r a g o ) Liu 1962) m o r p h o l o g i c a l l y a n d
physiologically r e lated to V i b r i o c h o l e r a e P a c i n i . It should
b e a p p a r e n t t h a t s o m e G r a m - n e g a t i v e r o d s with a p o l a r f l a g e l l u m and a single s o m a t i c c u r v e have b e e n c l a s s i f i e d i n
Pseudomonas Schroeter and Comamonas P a r k .
CONCLUSIONS
F i v e r e c o m m e n d a t i o n s , b a s e d on the o b s e r v a t i o n s p r e s e n t e d above, have been p r e p a r e d i n t h e i n t e r e s t of n o m e n c la t u r a L s t a b i li za t i o n ,
1 . Careful r e s e a r c h indicates that the infusorian generic
n a m e V i b r i o Mutter 1 7 7 3 , 39 h a s not b e e n u s e d i n the p r i m a r y zoological l i t e r a t u r e f o r m o r e than 100 y e a r s .
It i s
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recommended that Vibrio Mtiller 1773 be placed in the Offic i a l Index of Rejected Zoological Names.
2. P a c i n i (1854) was the f i r s t to d e s c r i b e a m i c r o b e which
i s recognized today a s a s p e c i e s of b a c t e r i a c u r r e n t l y c l a s sified i n the genus Vibrio. It i s recommended that the gene r i c name Vibrio P a c i n i 1854 be conserved against Vibrio
MUller 1773 and a l l other zoological n a m e s f o r the taxon to
avoid f u r t h e r changes i n the nomenclature of the genus, and
that i t be placed i n the List of conserved b a c t e r i a l g e n e r i c
n a m e s (Nomina g e n e r u m b a c t e r i o r u m conservanda).
3. P a c i n i (1854,411) d e s c r i b e d and named vibrio cholera
(sic). He w a s the f i r s t to u s e the g e n e r i c name Vibrio with
a validly published and legitimate specific epithet, c h o l e r a e ,
f o r the b a c t e r i u m which i s recognized a s the c a u s a l agent
of Asiatic c h o l e r a . It is recommended that Vibrio c h o l e r a e
P a c i n i 1854, the type species by monotypy, be placed i n the
list of n a m e s of type species of genera (Nomina generum
bacte r i o r u m cons ervanda)
.
4. It i s recommended that the specific epithet c h o l e r a e
i n the binary combination Vibrio cholerae P a c i n i 1854 be
placed i n the list of conserved specific epithets (Epitheta
spe cifica con s e rvanda)
.
5. It i s recommended that the National Collection of Type
Cultures s t r a i n 8021 (American Type Culture Collection
s t r a i n 14035) be recognized a s the neotype s t r a i n of Vibrio
cholerae Pacini 1854,
ACKNOWLEDGEMENTS
This work was supported i n p a r t by r e s e a r c h c o n t r a c t
DA 49-193-MD-2044 f r o m the United States A r m y Medical
R e s e a r c h a n d Development Command. The author i s deeply
indebted a n d grateful f o r t h e encouragement, counsel, and
generous a s s i s t a n c e of P r o f e s s o r H e r b e r t L. Ley, J r .
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