A STUDY OF THE SPECIES LACTOBACILLUS

PLANTARUM (ORLA-JENSEN) BERGEY

ET AL.1
CARL S. PEDERSON2
New York State Agricultural Experiment Station, Geneva, New York
Received for publication, November 5, 1935
In systematic work it is essential that the characters used to
define species shall be of such a nature that at least the majority
of the strains can be properly and readily identified. In only
relatively few cases have descriptions of species of bacteria been
sufficiently comprehensive to include even known variations and,
as a result, new strains or variants of species are frequently
described as new species. For such reasons it was found difficult
to arrange a key for the species of the genus Lactobacillus for
Bergey's Manual of Determinative Bacteriology (1934). Many
so-called species could not be separated from one another. This
situation was particularly evident in those species of non-gas-
producing organisms normally found in plant materials or in soil.
In a study of over four hundred strains of non-gas-producing
organisms isolated from fermenting substances no characters
were observed which could satisfactorily be used to separate the
cultures into groups that corresponded to described species.
It seemed, therefore, that nearly all of the descriptions that have
been given are not only too incomplete, but are also too restricted
in nature and that more inclusive descriptions should be used.
The description of the species Streptobacterium plantarum Orla-
Jensen (1919) syn. Lactobacillus plantarum Bergey et al., is the
1 Approved by the Director, New York State Agricultural Experiment Station
for publication as Journal Paper No. 109.
2 The author wishes to acknowledge the many helpful criticisms and sugges-
tions made by Drs. E. B. Fred and W. H. Peterson of the University of Wiscon-
sin, Dr. J. M. Sherman, New York State College of Agriculture, and Dr. R. S,
Breed, New York Agricultural Experiment Station.
217
JOBUNAL OF BACTZRIOLOGY, VOL. 31, NO. 3
218 CARL S. PEDERSON

first one given in this group that is sufficiently general to be

satisfactory.
The name was applied by Orla-Jensen to a series of strains of
bacteria isolated from butter, milk, cheese, fermenting potatoes,
beets, cabbage and dough. Complete fermentation characters
on forty-four strains were presented. Fermentation usually
varied somewhat with the source of nitrogen. Inactive lactic
acid was produced by the majority of strains, although a few
produced an excess of dextro-lactic acid. Studies on changes
produced in nitrogenous foods show that the strains do not split
the proteins to any degree. More or less complete descriptions
of other characters were given. The species differs but slightly
from Streptobacterium case Orla-Jensen which is described, in
contrast to Streptobacterium plantarum, as producing dextro-
lactic acid and as usually preferring lactose to maltose and
sucrose.
Variations in the sugar fermentations are the only characters
of a sufficiently constant nature to raise the question of establish-
ing more than one species in the group of cultures included under
the name Streptobacterium plantarum. When the results secured
by Orla-Jensen are summarized, it is found that if the group is
split, it should be on the basis of the fermentation of the sugars
xylose, arabinose, or raflinose. If these sugars are selected for
use in separating the strains into species, there would be as many
species as there are possible combinations of characters. Strains
that produce a vigorous xylose fermentation can be separated
into 67 per cent weak and 33 per cent strong arabinose fermenters,
or into 17 per cent weak and 83 per cent strong raffinose ferment-
ers. Strains of vigorous arabinose fermenters can be separated
into 74 per cent weak and 26 per cent strong xylose fermenters,
or into 24 per cent weak and 76 per cent strong raflinose fer-
menters. Variations are noted in the fermentation of these
sugars.
Over four hundred strains of non-gas-producing, short-rod
forms from various fermenting substances have been used in our
study. These studies have included morphology under different
growth conditions, growth characters, resistance to heat and to
 LACTOBACILLUS PLANTARUM (ORLA-JENSEN) BERGEY ET AL. 219

salt, hydrogen ion relationships, temperature relationships,

products of fermentation, and ability to ferment carbon com-
pounds. Morphology varied with conditions of growth and
under like conditions the majority of strains were the same.
Although other differences were noted, the greatest variation in
characters was found to occur in ability to ferment sugars.
Every gradation from a definite failure to ferment to a definite

60

s0

40

30

~20

FIG. 1A. FREQUENCY OF ACID PRODUCTION FROM VARIOUS CARBON COMPOUNDS BY

STRAINS OF LACTOBACILLUS PLANTARUM EXPRESSED AS PER CENT OF STRAINS
PRODUCING FROM 0 TO 9 CC. OF ACID IN 10 CC. OF MEDIUM
Ten spaces used for each sugar signify in the first block per cent of strains
producing no acid, the second, per cent of strains producing 0.1 to 1 cc. of acid,
the third space 1.1 cc. to 2.0, etc. The tenth space is used for percentage of strains
producing 8.1 to 12.5 cc.

positive fermentation was noted for nearly all of the carbon com-
pounds (fig. 1, a and b).
Almost all strains fermented glucose, levulose, mannose,
galactose, sucrose, maltose, lactose, raffinose and salicin, and to a
lesser extent, sorbitol, mannitol, dextrin and glycerol. The ma-
jority of strains did not ferment rhamnose, starch and inulin.
A few strains had little or no action on arabinose, while a greater
220 CARL S. PEDERSON

number had little or no action- on xylose. Melezitose was about

the only carbohydrate in which an approach to a clear-cut posi-
tive or negative fermentation was obtained. The same range of
variation occurred even where cultures were obtained from
identical materials.
If this group of cultures is divided into more than one species
on the basis of fermentation characters, the carbon compounds

so

FIG. 1B. FREQUENCY OF ACID PRODUCTION FROM VARIOUS CARBON COMPOUNDS

BY STRAINS OF LACTOBACILLUS PLANTARUM EXPRESSED AS PER CENT OF
STRAINS PRODUrCINGE FROM 0 TO 9 CC. OF ACID IN 10 CC. OF MEDIUM1
Ten spaces used for each sugar signify in the first block per cent of strains
producing no acid, the second, per cent of strains producing 0.1 to 1 cc. of acid,
the third space 1.1 cc. to 2.0, etc. The tenth space is used forpercentageof
strains producing 8.1 to 12.5 cc.
which could be used would be melezitose and arabinose and
possibly xylose and galactose; but when characters are studied
carefully, little justification can be found for dividing the group.
Even the fermentation of melezitose merely separates the less
active strains from the more active strains. Since this sugar
has not been used by previous workers and the separation has
little importance, it seems more reasonable to consider this as a
strain variation.
 LACTOBACILLUS PLANTARUM (ORLA-JENSEN) BERGEY ET AL. 221

Among the strains studied, cultures have been found which

agree in their characteristics with many of the closely related
so-called species described. The name Lactobacillus plantarum
(Orla-Jensen) Bergey et al. appears to be the most logical selection
of a name for this species, even though several strains had pre-
viously been described and named. However, the name men-
tioned is the first one given to a well described and inclusive
species. Described species which appear to be wholly or in part
identical are as follows:
Bacillus pabuli-acidi II Weiss3 is poorly described, and the
name given is in the form of a trinomial. The species was con-
sidered to be related to Bacillus a of von Freudenreich now called
Lactobacillus casei (Orla-Jensen) Holland. Possibly, strains
described by Weiss were related to Lactobacillus casei rather than
to Lactobacillus plantarum.
Six of the so-called species described by Henneberg in 1903
appear to belong to Lactobacillus plantarum. The descriptions
by Henneberg are very complete in certain respects, but incom-
plete in others. Descriptions differ but slightly, and variations
may possibly be dependent upon growth conditions. Henne-
berg's species differ from one another primarily in respect to their
fermentation of carbon substances. The first of these, to which
he has applied a trinomial Bacillus cucumerisfermentati was later
renamed Lactobacillus cucumeris by Bergey et al. in 1923. It is
the only well known name and the only one of the group of which
representative cultures are available. It is described as fer-
menting arabinose and dextrin vigorously. Two cultures re-
ceived from Henneberg show no significant differences from
cultures of Lactobacillus plantarum. Since the binomial Lacto-
bacillus cucumeris was proposed later than the binomial Strepto-
bacterium plantarum OrlaJensen 1919 (now Lactobacillus plan-
tarum), the latter name is the valid one. Descriptions of the
other species are similar. Bacillus Wortmanni is reported as
fermenting alpha methyl glucoside. Bacillus Listeri shows a
weak fermentation of arabinose. Bacillus Maerckeri shows a
3 Bibliographic references to all of these names will be found in Bergey (1934),
pages 307-312.
222 CARL S. PEDERSON
weak action on arabinose and alpha methyl glucoside but a strong
fermentation of dextrin. Bacillus Leichmanni II is an inulin
fermenter, a weak arabinose fermenter but a high acid producer.
Since it is described as granular, there may be some doubt whether
or not this should be regarded as identical with the species Lacto-
bacillus plantarum. Bacillus Leichmanni I and III either fail
or slightly ferment arabinose, lactose and raffinose, and do not
curdle milk. They seem to be more closely related to the species
Lactobacillus Delbrueckii described by Leichmann as Bacillus
Delbricki than to Lactobacillus plantarum. Possibly they are
intermediate between Lactobacillus plantarum and L. Delbrueckii.
Unfortunately the type of lactic acid formed was not determined
for any of these types. Bacillus Beijerincki is a weak fermenter
of arabinose and trehalose and has no action on lactose, mannitol
and milk. It is a weak strain more like B. Leichmanni I and III
and B. Delbricki in its sugar fermentations.
From this brief summary, it may be noted that species differen-
tations have been based primarily upon differences in sugar
fermentations, although in many cases these are not definitely
positive or negative.
Two other species, Lactobacillus conglomeratus Beijerinck (1901)
(1908) and Lactobacillus densus Beijerinck appear to be identical
with Lactobacillus plantarum, but the descriptions given are very
incomplete.
Three other names have been given to organisms that appar-
ently belong to this species subsequent to Orla-Jensen's descrip-
tion of Lactobacillus plantarum, namely Lactobacillus pentosus
and Lactobacillus arabinosus as described by Fred, Peterson and
Anderson and Bacterium busae-asiaticae as described by Tschekan.
The latter is apparently identical with Lactobacillus plantarum.
Lactobacillus pentosus and Lactobacillus arabinosus were described
shortly after the publication of Orla-Jensen's work and are more
properly regarded as varieties of this species, the differentiation
being based on the fermentation of xylose. In an unpublished
paper by Fred and Peterson (1935) complete biochemical char-
acterization is given of non-gas producing rods from fermenting
vegetables. In this manuscript, they have referred to the cul-
 LACTOBACILLUS PLANTARUM (ORLA-JENSEN) BERGEY ET AL. 223

tures by the names Lactobacillus plantarum and Lactobacillus

cucumeris.
CONCLUSION
It appears logical to retain the name Lactobacillus plantarum
as the species name for bacteria having the following char-
acteristics:
Lactobacillus plantarum (Orla-Jensen) Bergey et al.
Synonyms in whole or in part: Bacillus pabuli-acidi II Weiss,
Bacillus cucumeris fermentati Henneberg, Bacillus Wortmanni
Henneberg, Bacillus Listern Henneberg, Bacillus Maerckeri Henne-
berg, Bacillus Leichmanni II Henneberg, Bacillus Beijerincki
Henneberg, Lactobacillus pentosus Fred, Peterson and Anderson,
Lactobacillus arabinosus, Fred, Peterson and Anderson and
Bacterium busae-asiaticae Tschekan and probably Lactobacillus
densus Beijerinck and Lactobacillus conglomeratus Beijerinck.
Morphology: Non-motile, Gram-positive rods, ordinarily 0.7
to 1.0 by 3.0 to 8.0 microns, occurring singly or in short chains
with rounded ends. Under favorable growth conditions these
organisms tend to be short rods. Under adverse conditions
they tend to be longer; for example, in tomato juice agar at
450C. (Pederson, 1929). In fermenting vegetables, the organisms
tend to become longer as the acidity becomes greater. The
organisms are usually longer in milk than in broths. Differences
in morphology are well illustrated by Orla-Jensen.4
Gelatin colonies: No growth.
Gelatin-yeast extract-glucose stab.: Filiform growth. No lique-
faction.
Agar slant: Growth, if any, is very faint.
Broth: Turbid, clearing after a few days. A few strains
flocculate.
Litmus milk: Acid, usually coagulated.
Potato: No growth.
Indol not formed.
Nitrates not reduced.
The majority of strains form acid in glucose, levulose, mannose,
4 See plates 41 and 45, Orla Jensen, 1919.
224 CARL S. PEDERSON

galactose, arabinose, sucrose, maltose, lactose, raffinose, and

salicin, and to a lesser extent, in sorbitol, mannitol, dextrin,
glycerol, and xylose. Rhamnose, starch and inulin are usually
not fermented.
Lactic acid, usually inactive, with only small quantities of
acetic acid and carbon dioxide, are formed in the fermentation of
hexose sugars. Acetic and lactic acid are produced from the
pentoses. Forms up to 1.2 per cent acid in broth.
Salt tolerance: Usually grow in salt up to 5.5 per cent.
Microaerophilic.
Optimum temperature 30-35TC. Maximum 40TC. Mini-
mum 100C.
Thermal death temperature: 650 to 75TC. for 15 minutes.
Sources from which isolated: milk, cheese, butter, kefir, feces,
fermenting potatoes, beets, corn, chard, bread dough, sauerkraut,
cucumber pickles, tomato pickles, cauliflower pickles and spoiled
tomato products.
Habitat: Widely distributed in nature, particularly in ferment-
ing plant and animal products.
REFERENCES
BEIJERINCK, M. W. 1921 Archives N6erlandaises de Sciences Exactes et Natu-
relles Haarlem, S6rie II, Tome VII, 212-243. Also in Verzamelde Ge-
schriften van M. W. Beijerinck, Delft, 4, 54.
BEIJERINCK, M. W. 1908 Centr. fur Bakt., II Abt., 20, 1908, 641-650.
BERGEY, D. H. 1934 Manual of Determinative Bacteriology. 4th Ed. 664 pp.
Baltimore.
FRED, E. B., AND PETERSON, W. H. 1935 Manuscript.
ORLA-JENSEN, S. 1919 MWm. de l'Acad. Roy. d. Sci. et d. Lettres de Danemark,
Copenhague. Sect. d. Sci., 8 s6r., 5, No. 2.
PEDERSON, C. S. 1929 Tech. Bulls. 150 and 151, N. Y. State Agr. Exp. Station.