(SANITIZED)UNCLASSIFIED SOVIET PAPERS ON TAXONOMIC STUDIES OF THE MEMBERS OF THE GENUS SERRATIA(SANITIZED)
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.NTERNATIONAL BULLETIN OF BACTERIO~.Of'ICAL
NOr4FNCLATURE AND TAXONOMY
Volume 11 No. 1 January 15, 1961 pp. 7-12
THE TAXONOMIC STATUS OF
SERRATIA MARCESCENS BIZIO
(With the technical assistance of Z. Vyalou~lilovd)
Department of Microbiology
Faculty of Natural Sciences in Brno
Czechoslovakia
SUMMARY: 1. In our work we have studied mor-
phological, cultural and biochemical char-
acteristics of 68 strains of Serratia marces-
cens. 2. We propose that the genus Serratia
may have only one species. 3. We propose to
acknowledge strain BS 303 (ATCC 13880)as the
neotype culture of Serratia marcescens Bi~zio.
The oldest known species of the genus Serratia is Serratia
marcescens. It was named as early as 1823 and since this
time both its nomenclature and its taxonomy have under-
gone many changes. Recognition of this species as the type
species of the genus Serratia was proposed by Buchanan
(1918). It ie so recognized in all seven editions of the Ber-
gey~ s Manual.
The taxonomy of Serratia marcescens has been investig-
ated by Breed et al. 1924, 1927 , Topley and Wilson (1931)
and KrassilnikovT1949). The two latter authors place this
species in the genus Chromobacterium as Chromobacterium
prodigiosum. But the name Chromobacterium prodigiosum
is very little used. Much new information concerning the
biochemical and antigenic structure of Serratia marcescens
has been published by Davis, Ewing and Reavis 1957).
We have studied 68 strains secured from several collec-
tions underthe label Serratia marcescens. The list of strains
is given in Table 1. The methodauaed were those described
in our previous work (Martinet and Kocur, 1960). The
staining of flagella was made by Zettnow~ s method (Kabelilc
1925).
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_ q,,,. ~ _
INTERNATIONAL BULLETIN
301, 302, 303, 304, 813
731, 545, 544
147, 264
151
Institut of Plant Production, Prague
Institut of Epidemiology and
Microbiology, Prague
Prof. E. Steinhaus, Univ. of
California, Berkeley
400, 412, 414, 420, 454
455
361
375
376, 377
522
535
548
549, 580
619
620
689
696, 697
761
Culture Coll. of Entomogenous
Bacteria, Prague
C. B. van Niel, Stanford Univ.
Pacific Grove, California
Dept. of Agriculture, Ottawa,
Canada
Epid. and Hygiene Station, Brno
Department of Microbiol.
Hradec Kr31ov~
E. Eltinge, Mont Holl. Coll.
Massachusettes, U. S. A.
Dept. of Microbiology, Charles
Univ. Prague
Inst. of Biology, Czech. Acad.
Sci. Prague
Inst. of Microbiology, Univ. of
Tucuman
Dept. of Bact. Indiana Univ.
Bloomington
B. Hampl. Dept. of Biol. Sciences,
Prague
Dept. of Bact. Univ. of Queens-
land, Australia
Inst . of Fermentation, Sao Paulo,
Brazil
Dept. of Microbiol., Tech.
School, Bratislava
W.C. Haynes, NRRL, Peoria,
Illinois
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Page 9
BACTERIOLOGICAL NOMENCLATURE
AND TAXONOMY
Mor bolo The cells of all 68 strains of Serratia mar-
cescens studied are small Gram-negative rods occurring
individually or in groups. Their size varied from 0.8-1.0 x
1.5-1?. All the strains have motile cells.
Cultural characteristics. The colonies on nutrient agar
are circu ar, - mm in diameter, with slight undulate mar-
gins, slightly convex, smooth. Red pigment was produced
by moat strains. In nutrient broth all the strains developed
intense turbidity and formed alight sediment. Some strains
formed red rings.
Cultural and h siolo ical characteristics. The results
of the study of biochemical characteristics of 68 strains of
Serratia marcescens are given in Table 2.
Our study of 68 strains of Serratia marceacens brought
many interesting notions. As it can be seen from the tab-
ular summary No. 2, some strains showed different bio-
chemical features. The moat important variants here were:
Strain 780 alone did not hydrolyze casein; strains 548 and
619 did not reduce nitrates; strains 303, 304, 772 and 78T
showed positive hemolysis. It ie interesting that ihemajor-
ity+of these anomalies was observed in strains isolated from
different species of insects (e. g. strains 780, 772, 787).
Relatively great variability of some strains was observed
during the fermentation of carbohydrates. This problem
was not met only by us but also by Davis et al. (1957). Also
the differences in pigmentation are not surprising. On nor-
mal media and on media for pigmentation only 55 strains
pigmented at normal cultivating temperatures. Out of these
55 strains only 15 pigmented at 37'C; 13 strains did not
produce pigment at all.
The results obtained by us are essentially the same is
those given in Bergey's Manual (1957), in Krassilnikov'a
Key (1 X49) and agree with the data stated by Davis et al. (1957 ).
We do not agree, however, with Krassilnikov~ s ascrip-
tion of S. marcescens to the genus Chromobacterium solely
on the basis of its ability to produce pigment. We believe
that there are insufficient grounds for shifting this species
to a very different genus. As we have ascertained, Serratia
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. , 'Table 2. Results of biochemical tests with 68 strains of
Serratia marceecens
Test
Number
positive
Number
negative
Gelatin liquefaction
68
0
Casein hydrolysis
67
1
Nitrate reduction
66
2
Hydrogen sulfide I.
67
1
Hydrogen sulfide II.
0
68
Indole
0
68
Milk
68
0
Hemolysis
4
64
Glucose
68
0
Gas from glucose
43
25
Lactose
2
66
Sucrose
68
0
Maltose
63
5
Galactose
64
4
Fructose
67
1
Rhamno s e
0
68
Mannose
66
2
Inulin
0
68
)[ylose
6
62
Arabinose
0
68
Glycerol
66
2
Adonitol
64
4
Sorbitol
67
1
Mannitol
68
0
Ihilcitol
0
68
Starch
0
68
Eaculin
68
0
Koser~ s citrate
67
1
Simmons s citrate
66
2
Acetylmethylcarbinol
66
2
Methyl red
0
68
KCN
68
0
Phenylalanin
0
68
Catalase
68
0
Urease
0
68
Lipase
67
1
5'Ie NaCI
68
0
7.59E NaCI
47
l l
Pigment
55
13
Hydrogen sulfide I. = lead acetate papers
Hydrogen sulfide II. = modified Klingeri s agar
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BACTERIOLOGICAL NOMENCLATURE
AND TAXONOMY
marcescena differs from the representatives of the genus
Chromobacterium not only morphologically, but especially
biochemically and in the chemical structure of its pigment.
Since 1823 when the species was first described, authors
have described and named other new species. These new
epeciea were classified unsatisfactorily. In 1957 Davis et
al. (1957) concluded that the genus Serratia is monotypic,
including only one epeciea.
We have devoted much study to this problem and have
substantiated the findings of Davis et al. (1957) (Martinet
and Kocur, 1960).
? Owing to the fact that there exists no culture of the or-
iginal strain of Serratia marcescena, we propose the ap-
proval of our strain 303 as a neotype culture. The descrip-
tion of proposed neotype of Serratia marcescena:
Serratia marcescena Bizio (strain BS 303, ATCC 13880)
Small Gram negative rods, occurring individually or in
groups. The size ?of cells was 0.8 x 1.S?. Motile by
four lateral flagella.
Agar colonies (t days): Circular with slight undulate mar-
gin, 4-6 mm in diameter, slight convex, smooth, pro-
duce red pigment.
Gelatin colonies (5 days): Circular, smooth sinking in 2-3
days in gelatin, white.
Agar slant (1 day): Smooth, white, taking on an orange-red
to carmin red colour in 4-5 days.
Broth (1 day): Intense turbid and gray sediment.
Potato (2 days): Luxuriant growth, smooth, first white, later
red colour.
Peptone water with carbohydrates: Acid but no gas formed
from glucose, saccharoae, maltose, galactose, mannitol,
sorbitol, glycerol, mannoae and adonitol. Not attacked
by lactose, dulcitol, arabinose, rhamnose, xylose, in-
ulin.
Milk (2 days): Alcaline, coagulation sad peptonization.
Indole: -; HZS: -; VP: +; MR: -; nitrate +; catalase +; urease
-; lipase +; KCN +; phenylalanine -; Siinmons~ citrate +;
Koaer~ a citrate +; hydrolysis of gelatin +; casein +; hy-
drolysis of starch -; esculin +; hymolysis +.
Salt tolerance: Nutrient agar with 5'1e NaCI +; 7.5'16 NaCl +;
10'le NaCI ._.
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P+sQe 12
Aerobic .
Optimum temperature 30?C, minimum 10?C, maximum 37?
C.
Optimum pH 6.8, minimum 4.4, maximum 9. t.
Habitat: Isolated from pond water.
Breed, R. S. and M.E. Breed. 1914. The type species of
the genus Serratia commonly known as Bacillus prodigio-
sus. Jour. Batt. 9:545-557.
, 1927. The genus Serratia Bizio. Zentralbl.
f. Bakt. II Abt. 71:435-440.
. E. G. D. Murray and N. R. Smith. 1957. Bergey s
Manual of Determinative Bacteriology. 7th ed. 1094 pp.
Williams and Wilkins Co., Baltimore. Maryland.
Davis, Betty R.. W.H. Ewing and R.W. Reavis. 1957.
The biochemical reactions given by members of the Ser-
ratia group. Internl. Bull. Batt. Nomen. and Taxon.
7:151-160.
Kabelfk.J. 1915. Bakteriologick? technika drobnohledn?.
Praha.
Krassilnikov, N. A. 1949. OpredMit~l bakt~rij i aktino-
mycetov. Moskva.
Martinet, T. and M. Kocur. 1960. The taxonomic status
of Serratia plymuthica (Lehmann and Neumann) Bergey
et al. and of Serratia indica (Eisenberg) Bergey et al.
Internl. Bull. Batt. Nomen. and Taxon. 10(4):147-154.
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- INTERNATIONAL BULLETIN OF BACTERIOLOGICAL
` ~ NOMENCLATURE AND TAXONOMY
Volume 11 No. 3 July 15, 1961 pp. 87-90
CONTRIBUTION TO THE TAXONOMIC.
STUDIES OF SERRATIA KILIENSIS (LEHMANN
ET NEUMANN) BERGEY
(With the technical assistance of Z. VyslouEilovl)
Department of Microbiology
Faculty of Natural Sciences in Brno
Czechoslovakia
SUMMARY: A study of the morphological, cul-
tural and biochemical filatures of five strain?
of Serratia kil_i_e_n_sis (Lehmann et Neumann)
Bergey from different culture collections
leads to the conclusion that Serratia kilien-
sis ie a variety of Serratia marcescens
--- ------- ----------
In 1888 a red pigmentedmicroorganism, which was called
only bacterium h, was isolated from drinking water by Bzen-
ni~ in Kiel. The name Bacterium kiliense was given to it
later by Lehmann and Neumann 1896. Thi? species was
placed in the genus Serratia by Bergey (1923) with the name
Serratiakeilensis(sic Breed (1957)correctedthetpelling
of the specific epithet to kiliensis. Some authors (Lehmann
and Neumann 1927, Breed 1957, Davis et al. , 1957) have
suggested that Serratia kiliensis is a variety of or identical
with 5. marcescens. However, S. kiliensis is etillgivenspe-
cific statue by authors (Krasilnikov 1949. Breed 1957).
We have therefore undertaken to ascertain whethez S.
kiliensis and S. marcescens are synonyms.
We have studied 5 strains supplied as Serratia kiliensis
by various culture collections. Strain 274 was from the
Department of Agriculture, Ottawa, Canada; atrain300 from
Department of Biological Sciences, Purdue University,
Indiana; strain 526 from?J. Simpson, Prairie Regional Lab-
oratory, Saskatoon, Canada, strains 7462 and 922 from the
American Type Culture Collection.
The methods usedwere the same as in our previous work
(Martinet and Kocur 1960).
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Page 88
? INTERNATIONAL BULLETIN
Morphology. All five strains formed small Gram nega-
tive rods, singly or in groups. 0.8 x 1. 5-2.O?. All strains
were motile.
Cultural characteristics. Colonies were round with un-
dulate margin, slightly convex, smooth. On nutrient agar
two strains only were pigmented. In nutrient broth all
strains produced turbidity and sediment, two strains formed
a fragile pellicle. On potato, gelatin and on medium Dewey
and Poe (1943) only 2 strains were pigmented.
Biochemical characteristics. The ascertained biochemi-
cal characteristics are shown in Table 1.
No essential differences in morphological and cultural
characters were foundamong the five strains studied. Some
minor differences appeared in the biochemical characteris-
tics. Strain SZ6 hemolyaed blood and did not form gas from
glucose, strains 582, 274 and 581 fermented lactose. The
variability in fermenting lactose and producing gas. agrees
with the description given by Davis et al. (1957).
In other reepecteour results agree with the original des-
cription (1888) as well as with the data given by Krasilnikov
(1949) and Breed (1957). Breed (as well as some other
authors) suggests that S. kiliensie is probably identical with
Serratia marcescensbut does not prove thieidentity experi-
mentally. He stated that it is uncertain whether the micro-
organism isolated by Breunig(1888) was s strain of Serratia
marcescens not heavily pigmented. Our results confirm
this supposition. Breed also states that one of the features
differentiating S. kiliensie from S. marcescens is the fact
that S. kiliensie does not produce acethylmethylcarbinol.
Out of five strains studied only the strain 526 produced
acethylmethylcarbinol. Besides this difference, however,
we have ascertained no other difference between S. kiliensie
and S. marcescens. Comparison of our results with those
of Breed 1957), Davie et al. (1957) and with our earlier
work (Martinet and Kocur 1959] leads us to the conclusion
that S. kiliensie is only a variety of S. marcescens. This
conclusion is reached because no production of acethylmethyl-
carbinol is the only single one of the substantial biochemical
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Table 1. Reault? of biochemical teats with S atsaias
of S.rr_atia kilienais.
Numbes
Positive
Gelatin liquefaction
5
Casein hydrolyai?
5
Nitrate reduction
5
Hydrogen sulfide I.
5
Hydrogen sulfide II.
0
Indole
0
Milk
5
Hemolysis
1
Glucose
5
Gas from glucose
4
Lactose
3d
Sucrose
5
Maltose
4
Galactose ,
5
Fructose
5
Rhamnoee
0
l~fannose
4
Inulin
4
Xylose
4
Arabinose
0
Glycerol
4
Adonitol
0
Sorbitol
5
Mannitol
5
Dulcitol
0
Starch
0
Esculin
5
Koaert s citrate
5
Simmonat citrate
5
Ac ethylmethylca rbinol
1
Methyl red
0
KCN
5
Phenylalanin
0
Catalase
5
Urease
0
Lipase
5
596 NaCl
5
7.5'~i NsCI
4
Pigment
2
Hydrogen sulfide I.
lead acetate paper
Hydrogen sulfide II.
modified Klinger~s
d =delayed
agar
Nnmbez
Ne~ati~e
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_. w~ ~.M....g
features which is -suitable in taxonomic differentiation of the
Serratia genus. According to our opinion the mentioned bio-
chemical test ie amore substantial feature than the strain
differences in fermenting of carbohydrates or in pigmenta-
tion, etc. '
From our previous work (Martinec and Koeur, 1961) and
irompresent study it follows that the genus Serratia include~
only one species - S. marcescene with the variety Serratia
marcescens var. kiliensis.
Breed, R.S. and M.E. Breed. 1927. The genus Serratia
Bizio. 2entralbl. f. Bakt. II. Abt. T1:435-440.
E. G. Murray and R. N. Smith. 1957. Bergey s
Manual of Determinative Bacteriology. Williams and
Wilkins Co., Baltimore, Maryland.
Breunig. J. 1888. Bacteriologiache untersuchung des
Trinkwasserss des Stadt Kiel. Dlse. Kiel.
Davis, B.R. , W.H. Ewing and R.W. Reavis. 1957. The
biochemical reactions given by members of Serratia
group. Internatl. Bull. Bact. Nomen. and Taxon. 7;
151-~60.
Krasilnikov, N. A. 1949. Opr~delit~l bakt8?rij i aktinomy-
cetov. Moskva.
Lehmann, K. 8. and R.O Neumann 1886. Bakteriologi4che
diagnoetik. Munchen.
Martinec, T. and M. Kocur. 19b0. The taxonomic status
of S. plymuthica (Lehmann and Neumann) Bergey et al.
and of S. indica Eisenberg) Bergey et al. Internatl.
Bull. Bact. Nomen. and Taxon. ~4~ 247-t54.
1961. The taxonomic status of Serratia
marceecens Bizio. Internatl. Bull. Bact. Nomen. and
Taxon. 11 1): 7 -12.
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+ INTERNATIONAL BULLETIN OF BACTERIOLOGICAL
? NOMENCLATURE AND TAXONOMY
Volume 11 No. 3 July 15. 1961 pp. 73-7d
A TAXONOMIC STUDY OF THE
MEMBERS OF THE GENUS SERRATIA
(With the technical assistance of Z. VyslouiCilov[)
Department of Microbiology
Faculty of Natural Science in Brno
Czechoslovakia
SUMMARY: A study of the morphological, cul-
tural, and biochemical characteristics of it
strains of different species of the genusSer-
rati_a_indicates that most itzains of species
thus far recognized are identical with Ser-
ratia marcescens.
----- ----------
The names S. Piscatorum (Lehmann and
Neumann) Breed, S. YoseEtica (Fortinean)
Bergey, S. fu_sch_s_i_n_a ~Boeckhout et de Vries)
Bergey, S. anoliu_m_ Duran-Reynals et Clau-
sen) are to be regarded as synonyms of S.
marcescens Bizio and Serratia msriaorubra
----------- -------- _~~_M___
as a synonym of Serratia marcescens var.
-------- ----------
kiliensis.
The strains S. sa~onaria I and II Markov
should be removed from the genus Serratia
and placed provisionally in the genus Aleali-
genes_
For our studies of the species of the genus Serratiawe
have obtained from various collections cultures of strains
without designation of species or with obsolete names.
Among these cultures were strains names, e. g. Serratia
pyoseptica (Fortineau~ Bergey. Serratia anotium Duran-
Reynais and Clausen, Serratia fuchsina Boeckhout and de
Vries) Bergey and others. Breed 1948) regards most of
these names as synonyms of Serratia marcescens. while
Krasilnikov (1949) includes them in his Manual as names of
accepted species.
In the 1957 edition of Bergey s Manual neither of the two
new strains of Serratia, isolated and described by ldaskov
(1956) as Serratia sa onaria I and II, is included.
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Page 74
In ouz present work we have studied whether the several
aforesaid "species" have taxonomic significance and whether
some of them belong to the genus Serratia at all.
In our work we have thoroughly studied 1Z strains of
Serratia acquired from various collections. The list of
strains is given in Table 1.
The methods used were the same as in our previous work
(Martinec and Kocur, 1960).
Serratia anolium
S. anolium B 1700
S. pyoseptica SZ3
S. saponaria I. II.
S. fuchsina 150
S. marinorubra 318
S. piscatorum 415
Serratia sP. S 33, S Z6
ATCC 6065
W.C. Haynes, NRRL, Peoria,
Illinois
Dept. of Appl. Biol. NRC,
Ottawa, Canada
J. Simpson, Prairie Reg. Lab. ,
Saskatoon, Canada
W. Markov, Sofia, Bulgaria
B. Hsmpl, Dept. of Biol. Sci.
Prague
C. E. Zobell, La Jolla. Calif.
Inst. of Plant Product., Prague
J. Simpson, Prairie Reg. Lab.
Saskatoon, Canada
C.B. van Niel, Stanford Univ. ,
California
Inst. of Epidemiology and
Microbiol., Prague
Morphology. All studied strains were short. gram-nega-
tive rods occurring singly and in clumps. Size of individual
cells 0.8-1 x 1.5-Z.5?. All strains were motile except
Serratia saponaria II.
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BACTERIOLOGICAL NOMENCLATURE
AND TAXONOMY
Cultural characteristics. Colonies of most strains were
round, with undulate margin, slightly convex, smooth. Eight
strains were at first creamy or slightly rose colored, later
red, very slightly pigmented was S. saponaria I. and there
were three non-pigmented strains S. saponaria II. , no. 6065
and B 1700. In nutrient broth all strains produced turbidity
and sediment except S. saponaria I. and II. which grew very
poorly. S. ~eaponaria I. and II., however, grew well is
nutrient broth at pH 9. 6. The mentioned strains did not
grow on potato. On the medium recommended by Dewey and
Poe (1943) 6 strains only were pigmented.
Biochemical characteristics. The ascertained biochemi-
cal characteristics are shown in Table 2.
The results indicated that all the strains with the excep-
tion of Serratia saponaria I and II agreed with theeharacter-
istics of Serratia marcescene Bizio. The differenee? as
shown in Table 2 (lactose fermented by strain 318, gas not
formed from glucose by strains 523, 318, S 26, maltose not
fermented by strain S 4 and adonitolnot fermented by strain
527) are characteristic for Serratia marcescene. Serratia
marinorubra BS 318 (acethylmethylearbinol not formed is
identical with Serratia marcescene var. kiliensis.
We conclude that the aforesaid species S. anolium ~ran-
Reynals and Clausen, _S. pyoseptica (Fortineau Bergey,
Serratia fuchsina Boeckhout and de Vries) Bergey are to be
regarded as synonyms of Serratia marcescen? Bisio and
Serratia marinorubra as a synonym of Serratia maresscens
var. kiliensis.
Serratia marcescens differs significantly from Serratia
saponaria I and II. The differences in the biochemical
activity of these two'strains in comparison with the strain
Serratia marcescen? are so substantial, that we conclude
that these two strains should not be placed in the gsmts
Serratia. We have compared biochemical characteristics
of Serratia saponaria I and II with one strain of Alealigene?
taecalis 09 and with the characteristics of this species in
Bergey's Manual (1957). We have ascertained that bio-
chemical features of Serratia sa onaria I and II and Alcali-
~enea faecalie are very similar liquefaction of gelatin. no
fermentation of carbohydrates, growth at pH 9.6). On the
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- .. ~ -_ ~~. ~ ~ _T .
wl c o oI ~I ~I ~~ ~I ~I I
~I vJt yl ~I yil hl hl yil ~I ~i~ t~~
Gelatin liquefaction + + + + + - + + + + + +
Casein hydrolysi^ + + + + - - + - + + + +
Nitrate reduction + + + + - + + + + + + +
Hydrogen sulfide I. + + + + - _ + + ? + +
Hydrogen sulfide II. +w +w - - - - - - - - - -
Indole - - - - - - - - - - -
Milk + + + + - - + + f + + +
Hemolysis - - - - - - - - - - - -
Gtueose + + + + - _ + - + - - +
Gas from glucose + + + - - - + - + - - +
Lactose - - - - - - + - - - -
Suezose + + + ' + - - + + + + + +
Maltose + + - + - - + + + + + +
Galactose + + + + - _ + + + + + +
Fructose + + + + - - + + + + + +
Ithamnose - - - - _ - - _ _ -
IYtanilOee + + + + - - + + + + + +
lnulin - - - - - - - + - - - -
Xylose - - - - - - - - - - - -
Azabinose - - - - - - - - - - -
Glycerol + + + - - _ + + + + + +
Adonitol + + + - - - - + + - -
Sorbitol + + + + - - + - + + + +
Mannitol + + + + - - + + + + + +
Thtlcitol - - - - - - - - - - - -
Starch - - - - - - - - - - - -
Eseulin + + + + - - + + + + + +
Koser~ a citrate + + + + - - + + + + + +
Simmons' citrate + + + + - - + + + + + +
Aeethylmethylcarbinol + + + + - - + - + + + +
Methyl red - - - - - - - - - - - -
KCN + + + + + - + + + + + +
Phenylalanine - - - - - - - - - - - -
Catalase + + + + + + + + + + + +
Urease - - - - - + - - - - - -
Lipase + + + + - - + + + + + +
5'-s NaCl + + + + + + + + + + + +
7. S'R NaCI + + - + + + + + + + + +
Pigment - + + + - + + + - - -
w =weak
Hydrogen sulfide I. =lead acetate paper
Hydrogen sulfide II. =modified Kliger medium
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BACTERIOLOGICAL NOMENCLATURE
AND TAXONOMY
basis of this observation we recommend placing Serratia
saponaria I and II provisionally in the genus Alcali;eves.
The definitive placing of this species will be possible only
after having compared it with other species of the genus
Alcaligenea.
Special attention should be given to the species S. pisca_
torum (Lehmann and Neumann) Breed. This species has
been recognized both by Krasilnikov(1949) and Breed (1957).
But this species is not deposited in any collection and there-
fore it cannot be compared with Serratia marcescens. The
only strain assigned to S. piscatorum, which we could com-
pare, was isolated several years ago from soil (see Table
1). We have ascertained that this strain is identical with
Serratia marcescens. On comparison of the original ehar-
acteriatics 1884 as well as the description in Bergey's
Manual (1957) with the features of Serratia marcescens, we
find only small differences. The faetthatS. piscatorum Caa
produce pigment at 37? and that this pigment is soluble in
water, cannot be taken for such an essential feature, on the
basis of which it would be possible to acknowledge S_. isca-
torum a valid apeciea, because this feature can be fonod
also in some strains of Serratia marcescens. We have
ascertained that out of 68 strains studied at 37 ?C only 15
strains produced pigment (unpublished data).. Also the fact
pigment of some heavily pigmenting strains of Serratia mar-
cescens is soluble in water, was stated already in 19'18 by
Breed and recently it has been experimentally proven by
Williams et al. (1958). For these reasons we regard Ser-
ratia piscatorum (Lehmann and Neumann) Breed as a syn-
onym of S. marcescens.
4: 487-501.
Breed, R. S. and M, E. Breed. 1917. The genus Serratia
Bizio. Zentralbl. f. Bakt. II. Abt. 71: 435-440.
E. G. D. Murray and A. P. Hitchens. 1948. Ber-
gey' s Manual of Determinative Bacteriology. Williams
and Wilkins Co., Baltimore, Maryland.
and R. N. Smith. 1957. Bergey's Manual of
Determinative Bacteriology. Williams and Wilkins Co.,
Baltimore, Maryland.
Boeckhout, F. W.J. and J.J. Ott de Vries. 1898. tJber
einem neuen chromogenen Bacillus. Zentralbl. f. Bakt.
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? p:g. Ta
Davis. B.R., W.H. Ewing and R.W. Reavis. 1957. The
biochemical reaction given by members of the Serratia
group. Interaatl. Bull. Bact. Nomen. and Taxon. T:
151-160.
Du Bois, Saint Sivrin. 1894. Panaris des pbcheurs et
microbe rouge dale sardine. Ann. Inst. Pasteur.
8: 152-160.
Duran-Reynals and H.J. Clausen. 1934. A contagious
tumor-like condition in the lisard (Anolis equestris) as
induced by new bacterial species, Serratia anolium (sp.
n. ). Jour. Bact. 33: 369-3T6.
Fortinesu, L. 1905. L~ Erythrobacillus pyosepticus et lea
bactbries rouges. Bull. Inst. Pasteur. 3: 13-14.
Lehmann, K. B. and R.O. Neumann. 1886 Bacteriolog-
ische Diagnostik. Munchen.
Markov, W. 1956. Serratia saponaria I. and II. Schid-
liche Bakterien in der Seife. Zentralbl. f. Bakt. II.
Abt. 110. 26-32.
Martinee, T. and M. Kocur. 1960. The taxonomic status
of Serratia plymuthiea (Lehmann and Neumann) Bergey
et sl. and of Serratia indica (Eisenberg) Bergey et al.
Internatl. Bull. Bact. Nomen. and Taxon. 10(4): 247-254.
1961. The taxonomic status of Serratia
marce cans Bizio. Interriatl. Bull. Bact. Nomen. and
Taxon. 11 1): ? -12.
Williams, R.P. , W.W. Taylor, D. Hawkins and I. L. Roth.
1958. A water soluble diffusible pigment produced by a
strain of Serratia marceacena (Chromobacterium prodig-
iosum) Nature. 182: 1028-1019.
ZoBell, C.E. and H.C. Upham. 1944. A list of Marine
Bacteria including descriptions of sixty new species.
Bull. Scripps. Inst. Oceanography, La Jolla. 5: t55.
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INTERNATIONAL BULLETIN OF BACTERIOLOGICAL
' NOMENCLATURE AND TAXONOMY
Volume 10 No. 4 October 15, 1960 pp. 247-Z54
THE TAXONOMIC STATUS OF
SERRATIA PLYMUTHICA (LEHMANN AND
NEUMANN) BERGEY ET AL. AND OF SERRATIA
INDICA (EISENBERG) BERGEY ET AL.
(With the technical assistance of Z. Vyslou~ilov~)
Department of Microbiology
Faculty of Natural Sciences
Brno, Czechoslovakia
Serratia plymuthica is one of the commonly recognized
species of the genus Serratia. This microorganism, first
isolated by Fischer (1887), was named Bacterium plymuthi-
cum by Lehmann and Neumann (1896) who concernedthem-
selvee with its classification. These authors showed that the
characteristics of this species allied it closely with Serratia
marcescens, perhaps it is quite identical. It has been given
species status rather uncritically even in such recent man-
uals on taxonomy as that of Krassilnikov 1949 and in Ber-
gey' s Manual, ed. 7, 1957.
Inasmuch as neither the original description of Serratia
1 muthica or the characterizations given by Krassilnikov
1949) and Breed(1957) are complete on the basi^ of present-
day needs, we have studied critically the justification for
the recognition of this as a distinct species.
Serratia indica (Eisenberg) Bergey et al. was first iso-
lated in 1884 by Koch from the alimentary tract of a Java
ape. The organism was not named by Koch, thebinomial-
Bacillus indicus Eisenberg-was given by Eisenberg(1886).
The history of the discovery of this species, the nomen-
clature and the justification of its validity was worked out by
Breed (1926).
Recently there have appeared opinions that this species
is identical with Serratia marcescens, e. g., Breed (1957)
thinks that it represents the R-form of S. marcescens.
Davis et al. (1957), as the result of a study of 50 strains of
the genus Serratia, concluded that this genus has only one
species, i. e. S. marcescens.
In this contribution we are able experimentally to verify
the hypothesis of Davie et al.
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Q., ~
We have studied 6 strains under the name of Serratia
plymuthica, secured from several collections: strain 192
from Prof. J. De Ley, Brussels, strains 139 and 162 from
Department of Bacteriology, Indiana University, Blooming-
ton, Indiana, strain 183 from the American Type Culture
Collection, strain 847 from Prof. van Niel, Pacific Grove,
California and strain 497 from Dept. of Agric. Ottawa.
We have also studied in detail 10 strains acquired from
various collections under the name Serratia indica. The
origin of these strains is given in Table I.
Serratia indica
Strain No. Source of Strain
305 Own isolates
T72 Research Institute of Plant Produc-
tion, Prague
33 and 135 Dept. of Bacteriology, Purdue
University, Indiana
Bu 209 Biol. Inst. Czech. Acad. Sci. , Prague
341 W.C. Haynes, Northern Regional
Res. Lab. , Illinois
IZ 358 Institute of Fermentation, Sao Paulo,
Brazil
447 Dept. of Agriculture, Ottawa, Canada
4002 and 4003 American Type Culture Collection
We have studied the morphological, cultural, and bio-
chemical characteristics of strains of both S. plymuthica
and S. indica, using the following methods: Grams s stain as
modified by Hucker, motility was detected by the Hajna
method, gelatin liquefaction was testedby Frazier s method
and by ^?ab inoculation on 15^/o gelatin, nitrate reduction by
Grier-Illosway~ s agent, hydrogen sulfide by means of lead
acetate strips and in modified Klinger agar Davis et al.
1957), indole by Kov~cs~ reagent.
The fermentation of sugars was studied in peptone water
with 1'lo sugar, starch hydrolysis was tested by Lugol solu-
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e R > rPage 249
BACTER~0ItiO81C11f`I;'I~OMENCLATURE
AND TAXONOMY
tion, and the utilisation of citrate determined in Koser' ? and
Simmons' medium. Production of acetylmethylcazbinol we
ascertained by Voges-Proskauer's medium by Leifson's
reagent (Methods for Pure Culture Study of Bacteria (1946)) .
Production of phenylpyruvic acid from phenylalanine and
growth in presence of KCN were studied by the methods
recommended by the Enterobacteriaceae Subcommittee
(1958). Urease we ascertained by Christensen'? method
(1946) and by Cowan's microtest (1952), lipase by Bulder's
method (1955). Pigment production was noted on potato.
nutrient agar, gelatin and on the medium recommended by
Dewey and Poe (1943).
Morphology: All six strains of S. plymuthies formed
small Gram-negative rods occurring individually or in
.groups. The sire varied from 0.8-1.0 x 1.5-2.O?. All
the strains were motile.
Cultural characteristics: The colonies on nutrient agar.
circular with undulate margins, slightly convex, smooth.
Red pigment was produced only by the strains 183 and 847,
the other were cream-colored. In nutrient broth all the
strains developed intense turbidity and formed a slight sedi-
ment.
Biochemical characteristics: The results of the study of
biochemical characteristics of both S. plymuthiea and S.
indica are given in Table 2.
Discussion. We wished to determine the taxonomic status
of the species S. plymuthica (Lehmann et Neumann) Bergey
et al. , by testing Lehmann arid Neumann' s hypothesis that
it might well be identical with the species S. marcescens
Bisio. In a study of the morphological and cultural charac-
teriatics of S. plymuthica we found no significant differences
among the six strains. Differences occurred only in pig-
ment production (3 strains formed no pigment). The differ-
ences in biochemical characteristics were a? follows: One
strain did not liquefy gelatin or hydrolyse casein. 5 strains
showed delayed fermentation of lactose. t strains fermented
inulin, I strainfermented adonitol. We attach no importance
to these differences. Other biochemical characteristics
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_ Table 2. Results of biochemical tests with 6 strains of
? ~ Serratia plymuthica and 10 strains of Serratia
indica.
S. plymuthica S. indica
No. + No. - No. + No. -
Gelatin liquefaction
5
1
10
0
Casein hydrolysis
5
1
10
0
Nitrate reduction
6
0
10
0
Hydrogen sulfide I
6
0
10
0
Hydrogen sulfide II
0
6
2w
8
Indole
0
6
0
10
Milk
6
0
-10
0
Hemolysis
0
6
0
10
Glucose
6
0
10
0
Gas from glucose
6
0
5w
5
Lactose
5d
1
0
10
Sucrose
6
0
10
0
Maltose
6
0
10
0
Galactose
6
0
10
0
Fructose
6
0
10
0
Rhamnose
0
6
0
10
Mannoae
6
0
10
0
Inulin
2
4
0
3 0
Xylose
0
6
0
10
Arabinose
0
6
0
10
Glycerol
6
0
30
0
Adonitol
1
5
10
0
Sorbitol
6
0
10
0
Mannitol
6
0
10
0
Dulcitol
0
6
0
l0
Starch
0
6
0
10
Esculin
6
0
10
0
Koser~ s citrate
6
0
10
0
Simmons citrate
6
0
10
0
Acetylmethylcarbinol
6
0
10
0
Methyl red
0
6
0
10
KCN
6
0
10
0
Phenylalanine
0
6
0
10
Catalaee
6
0
10
0
Urease
0
6
0
10
Lipase
6
0
10
0
5'Jo NaCI
6
0
10
0
7. 5'/o NaCl
6
0
10
0
Pigment
3
3
10
0
Hydrogen sulfide I =lead acetate papers. w =weak.
Hydrogen ^ulfide II: - modified Klin~er~ s agar. d =delayed.
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Page Z51
BACTERIOLOGICAL NOMENCLATURE
AND TAXONOMY
agree with the work of Davie et al. (1957).
Our results agree to some extent with Fischer a des-
cription (1887)in which the difference between S. plymuthica
and S. marcescene is said to lie in the fact that the cells of
S. plymuthica are thicker than tl.cse of S. marcescene, and
that a crimson pigment is produced only by S. plymuthica.
Neither these difference^ or those recognized by Breed
seem adequate today. .Breed regarded as the main differ-
ence between these species the production of gas from some
eugare by S. plymuthica, especially from glucose. This is
also true of S. marcescene (Bergey s Manual 1957), as was
found also by Davis 1957 and by us. In our opinion it is
not advisable to use the ability of the genus Serratia to pro-
duce gas from eugare as a taxonomic criterion for distin-
guishing species; we regard it as a variable characteristic.
Having compared the characteristics of the species Ser-
ratia plymuthica with the results of Davis et al. (1957) and
with the characteristics of Serratia marcescene (in Bergey e
Manual(1957) as well as with the characteristics of 68 strains
studied by us, we came to the opinion that Serratia Plv_
muthica is a junior synonym of Serratia marcescene.
Morphology: All strains of Serratia indica were mor-
phologically identical, small rode, occurring singly and in
chains; size of individual cells 0.8-1.0 x l.8?. Cells were
motile and Gram-negative.
Cultural characteristics: Colonies were roundwith slight
undulate margins, smooth, some strains were rough. Most
strains at first were creamy or slightly rose-colored, later
red. In nutrient broth all strains produced turbidity and
white sediment. On potato and in medium recommended by
Dewey and Poe (1943) all strains were pigmented.
Biochemical characteristics: The ascertained biochem-
ical characteristics are shown in Table 2.
Discussion. The task of our present work was to secer-
tain experimentally whether these strains of Serratia indica
were identical with Serratia marcescene. We conclude that
there are no substantia erencee between the individual
strains of Serratia indica studied. Some differences in re-
sults appeared in the production of hydrogen sulfide as de-
termined by tw~znet]iods, . ;Sy,use of the first method (lead
R ^'? 4" ~x
.. e F ~ ,~ a
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1 1$s [h )l t`:D Y ~ ~... _ a Sa .C W.F' ~~ `~.J; L ,";
acetate paper strips) we found sulfide produced by all ten
strains, with the second method (modified Klinger s agar)
we ascertained weak production in 2 strains only. These
results agree with those of other authors (Clarke 1953.
Davis et al. 1957).
Certain differences between individual strains were noted
in the production of gas from glucose. Five strains out of
the total of 10 produced a small amount of gat. We do not
attach great importance to this difference. Similar results
were obtained by Davis et al. (1957).
Only a small part of our results can be compared with
those recorded in the original description by Eisenberg
(1886) which included morphological and cultural character-
istics only. Our results agree with the characteristics re-
corded by Eisenberg..
Breed (1957) described Serratia indica more fully buY
emphasized the morphological and cultural characteristics
without adequate consideration of the biochemical character-
istics. He does not list the kinds of sugar that this species
utilizes. On the basis of Reedy swork(1937) Breed suggests
that Serratia indica is probably an R-form of Serratia mar-
cescens. Our results in part verify this hypothesis.
Kraseilnikov (1949) in hie Guide gives no new data a-
bout this species, he places it in the genus Chromobac-
terium.
We attach the greatest importance to the comparison of
our work with the results of Davie et al. (1957). On the
basis of the study of 50 strains of genus Serratia, these
authors formulated the hypothesis that this genus has only
one species. We have proved experimentally that this hy-
pothesis is correct. We haveaecertainedthatthecharacter-
istics of 10 strains of Serratia indica agree not only with
the results of the authors mentioned above but alto with the
characteristics of 68 strains of Serratia marcescens, which
we have also studied in detail.
From our study of the .morphological. cultural and bio-
chemical character of 68 strains received as Serratia
marcescens Bizio. 6 strains as S. plymuthica (lshmann
and Neumann) Bergey et al. and 10 strains as S. iadica
(Eisenberg) Bergey et,al., we conclude that the species
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Page 253
BACTERIOLOGICAL NOMENCLATURE
AND TAXONOMY
names S. plymuthica and S. indica should be regarded as
junior synonyms of S. marceacene.
Breed, R.S. and M. E. Breed. 1924. The type species of
the genus Serratia commonly known as Bacillus rp odig_
ioaus. Jour. Bact. 9: 545-557.
1926. Serratia indica Bergey, the red chromo-
genic bacterium originally isolated by Koch in India.
Jour. Bact. 11: 76-77.
1927. The genus Serratia Bizio. Zentral.
f. Bakt. II. abt. 71: 435-440.
E. G. D. Murray and N. R. Smith. 1957. Bergey s
Manual of Determinative Bacteriology, 7th ed. 1094 pp.
Williams and Wilkins Co., Baltimore, Maryland.
Bulder, C. J. E.A. 1955. Some observations on the lipo-
lytic activity of microorganisms and a new method for
its detection. Antonie van Leeuwenhoek 21: 433-445.
Christensen, W.B. 1946. Urea decomposition as a means
of differentiating proteus and paracolon cultures from
each other and from Salmonella and Shigella types.
Jour. Bact. 52: 461-466.
Clarke, P. K. 1953. Hydrogen sulphide production by
bacteria. Jour. Gen. Microbiol. S: 397-497.
and S.T. Cowan. 1952. Fermentations: Biochemical
micromethods for bacteriology. Jour. Gen. Microbiol.
6:784-794.
Davis, Betty R., W.H. Ewing and R.W. Reavis. 1957.
The biochemical reactions given by members of the
Serratia group. Intern. Bull. Bact. Nomen. and Taxon.
7: 151-160
Dewey, B.T. and Ch. F. Poe. 1943. A simple artificial
medium for pigment production by members of the genus
Serratia. Jour. Bact. 45: 485-498.
Eisenberg, J. 1886. Bakteriologieche Diagnoatik. Ham-
burg and Leipzig.
Enterobacteriaceae Subcommittee. 1958. Report of the
Enterobacteriaceae Subcommittee of the Nomenclature
Committee of the International Association of Micro-
biological Societies. Intern. Bull. Bact. Nomen. and
Taxon. 8: 25-70.
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Kocur, M. and T. Martinet. 1957. O v~skytu pigment-
uj~c~ch bakterif ve vod~ch KnfniLrakc~pi~ehrady. Publ.
Fac. Sci. Univ. Masaryk. Bruno, No. 388, 1-12.
Krassilnikov, N. A. 1949. OpredLlit2~1 baktl~rij i aktino-
mycetov. Moskva.
Lehmann, K. B. and R.O. Neumann. 1896. Bakteriologiache
Diagnoatik. Munchen.
Manual of Methods for Pure Culture Study of Bacteria. 1946.
Geneva, New York.
Reed, G.B. 1937. Independent variation of several char-
acteristics in Serratia marceacens. Jour. Batt. 34:
255-266.
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