RS,
Ba
Keates

CLASSIFICATION OF DAIRY BACTERIA

By H. W. CONN, W. M. ESTEN and
W. A. STOCKING.

From the Report of the Storrs (Connecticut) Agricultura

Experiment Station for 1906

CLASSIFICATION OF DAIRY BACTERIA

By H. W. CONN, W. M, ESTEN and
W. A. STOCKING

From the Report of the Storrs (Connecticut) Agricultural

Experiment Station for 1906

A CLASSIFICATION OF DAIRY BACTERIA.

BY H. W. CONN, W. M. ESTEN AND W. A. STOCKING.

In the Annual Report of this Station for 1899 there was pub-
lished a preliminary report upon the Classification of Dairy
Bacteria as they had been studied in our laboratory. This has
been up to the present the only attempt made at a systematic
arrangement of these bacteria and it has been used somewhat
extensively by bacteriologists. The large amount of work that
has been done since that time and the great changes that
have occurred in the methods of description and diagnosis of
bacteria have made that paper no longer a correct representa-
tion of the present knowledge of dairy bacteria. Requests for
that paper are still occasionally received, but inasmuch as it
is so far from representing the present state of the known facts
it is no longer regarded as advisable to distribute the paper
in question. For this reason, as well as because of the accu-
mulation of a large amount of further information, it is thought
wise to publish at the present time a second paper upon the
same subject, bringing the facts up to date. The previous
paper has served a purpose. It was recognized when it was
published, that it was only a temporary expedient, designed
as a stepping stone toward something better. It is hoped that
the present may serve a better purpose in the same direction
and bring us nearer to a clear, satisfactory classification of
dairy bacteria.

Since the publication of the previous paper we have done a
large amount of work upon these bacteria. Most of it has been
done in the laboratory of Wesleyan University by H. W. Conn
and W.M. Esten. A considerable portion has been done in the
laboratory at Storrs by W. A. Stocking and this paper is there-
fore the combined results of the two laboratories. The actual
writing of the paper and the arrangement of the scheme of
classification has been done by H. W. Conn who is therefore

92 STORRS AGRICULTURAL EXPERIMENT STATION.

responsible for the classification and arrangement of groups
hereby adopted. It will be seen in the following pages that a -
large number of new forms have been added to those described
in the previous paper and that not a few described previously
have been excluded from the present list as insufficiently
known. Our work in the last few years has added a large
number of new forms to our list. These types which we have
studied have been derived from a variety of sources. The
larger part of them have been obtained from milk and milk
products, either in the vicinity of Middletown or Storrs, and
are therefore especially Connecticut forms. We have, however,
received many cultures from elsewhere. Dr. Harding of the
Geneva Station has kindly sent a large number; Harrison of
Guelph, Canada, has sent us some; several have been sent by
Marshall from Michigan; a large number were furnished us from
the Board of Health Laboratory of New York City and sepa-
rated from New York City milk which is obtained from a wide
territory in the vicinity of that city; some have been sent from
Dr. Weigmann of Germany; some from Switzerland by Freud-
enreich; some from Italy by Gorini; and a few isolated forms
have come from other sources. The cultures that we have had
for examination have thus come from a very wide territory and
while by no means complete, they represent fairly the dairy
forms in the parts of the civilized world where bacteria are
studied. Of the dairy forms described in the following pages
the large majority we have ourselves had the opportunity of
studying in our laboratory. But in order to make this list of
dairy forms as complete as possible we have in the following
pages inserted with the types that we have ourselves studied,
the descriptions of quite a number that we have not seen but
which have been well described by others. This has been done,
however, only when the published descriptions are fairly com-
plete and are sufficient to make it possible to determine the re-
lations of the types in question. There have been a large num-
ber of bacteria mentioned and partly described in literature in
regard to which this is impossible. Many bacteria have been
mentioned as occurring in dairy products, and many named,
whose description is so meagre that it is absolutely impossible
for anyone to recognize them. In the Manual of Swithenbank
these names have been collected and the descriptions given.

CLASSIFICATION OF DAIRY BACTERIA. 93

But most of them are worse than useless at the present time,
for no one would be able to identify the bacteria from these
meagre descriptions or even to tell whether the bacteria des-
cribed belong to one or another group of bacteria. Ina few
cases this is possible, and where we have been able to do so we
have associated thése forms with some of the groups that we
have described more in detail below. But the value of the
descriptions in earlier literature varies with the completeness
of the description, and the descriptions of a majority of these
earlier described bacteria are so meagre that they are absolutely
worthless. When, therefore, a type of bacterium has been
named from characters which will apply to a score or more
different types there is nothing to do but neglect it entirely.
Not even its name can be retained if the type cannot be recog-
nized. We have therefore, left out of our list large numbers
of these types upon the ground that they must hereafter be
absolutely omitted from descriptions because of the impossi-
bility of determining what they are. We have tried to include
in our list all bacteria that have been sufficiently described to
render their recognition fairly sure, but, of course, there may
be some omissions.

In our recent work we have tried to include in our technical
routine of description all of the characteristics usually adopted
for the general description of bacteria. We have, however, not
adopted the practice of determining the formation of indol or the
reduction of nitrates. The reasons for this omission have been
that the data in question have not seemed to us to have much
significance in relation of dairy problems with which we have
been particularly concerned. Since these data have not been
determined in the laboratory routine they have, of course, not
been included in the following descriptions. It will be noticed
that with some of the forms described by us the description is
not complete. This is especially true of some of the earlier de-
scriptions of types which have not been found in abundance
and have not reappeared in our later work. Many of our
earlier described species, however, though incompletely de-
scribed at the time, associate themselves so closely with the
forms more recently studied that they are merged in groups,
which we now recognize and of which we give a more complete
description. A few forms, however, are still left with nothing

94 STORRS AGRICULTURAL EXPERIMENT STATION.

except the original description made several years ago, a de-
scription which may be too incomplete to be of very much value.
These are in a few cases retained in the following pages, but
only for species that possess very striking characteristics. For
instance, MZ. 1. rubzdus, having the power of producing a bril-
liant red pigment, is so striking a type that we did not think
it wise to omit it even though the description was based upon
a single culture obtained several years ago and is more or less
. incomplete.

It will manifestly require many years before a complete
knowledge of dairy bacteria can be obtained. While we have
in the following pages described about one hundred and sixty
types, we recognize that this is by no means the complete list
of even common forms; but to complete the list will require
many years of laborious study. When we recognize the extreme
variability of bacteria types, especially in physiological charac-
ters upon which their classification may be partly based, it be-
comes evident that to reach an end of the description of dairy
forms is almost an impossibility. Our recognized types, as will
be shown, vary in all directions and run into each other by more
or less complete intermediate links. Every new set of cultures
which we obtain from even similar sources show variations in
various properties connecting them more or less with other
types. This has been forced upon us more and more, as the
data we have collected has increased, until we have almost con-
cluded that the task of arranging these forms into species or
even groups is hopeless; for even at the very best these groups
will show such wide internal variations as to connect them
more or less completely with the closely associated forms.
Before such groupings can be finally made an immense amount
of work and many years’ study must be given. But every
attempt to formulate our ideas clears our conceptions, and
hence it is thought that the present endeavor to arrange the
dairy bacteria in a scheme of classification, even though of
necessity incomplete and doubtless requiring later changes,
will bring us nearer to a proper conception of their relation and
help toward a real understanding of problems of bacteriology.

The question of species —The question of species among bac-
teria is at present an insoluble puzzle. It has become manifest
that it is quite impossible to carry over to the classification of

CLASSIFICATION OF DAIRY BACTERIA. 95

bacteria the conception of word species which zodlogists and
botanists have developed in the last century. It has been
recognized by the modern zoologist that the early conception of
a species, as something sharp and distinct, is bound to be modi-
fied as variations are recognized. If this is true in animals and
higher plants, it is more emphatically true of bacteria. Indeed
we must practically abandon any thought of using among bac-
teria the term species with a meaning which has any similarity
to that which is used for the rest of the living world. The
question as to whether physiological variations are sufficient to
characterize a new species is one which we cannot now answer,
but this would be involved in an attempt to determine species
of bacteria. Into these questions we do not propose to enter at
all and shall not make any endeavor to arrange bacteria descrip-
tions in the form of species and genera. The term species, there-
fore, will not be used at all in the following classification.

As our data has been increasing, it has become more and more
evident that the bacteria which we have been studying natur-
ally fall into series of groups. This has been dimly recognized
by bacteriologists for some years, the &. coli group, for ex-
ample, being one that for many years has had a somewhat
recognized position. As we have collected the dairy bacteria,
we have been able to see such groups becoming more and
more apparent. In such a group there is usually a somewhat
central type, but from this the numerous cultures which may be
obtained and compared with each other show variations in all
directions. Inasmuch as our characters are chiefly physio-
logical, the variations of physiological properties naturally
produce an immense confusion in the attempt to satisfactorily
arrange cultures in groups. By these variations our different
groups are more or less connected with each other, even when
the groups are founded upon clear, sharp, positive distinctions.
For example, the power to liquefy gelatin has already been
recognized as one of the most fundamental characters, and yet
this power of liquefaction is manifestly subject to wide varia-
tions. As has been shown several times, organisms which
have the power of liquefying gelatin may lose this power more
or less completely, and hence it is perfectly clear that some
groups which liquefy may perhaps be only physiological modi-
fications of non-liquefying types. Among the cocci described

96 STORRS AGRICULTURAL EXPERIMENT STATION.

in the following pages it will be seen that there is a white
liquefying anda white non-liquefying group. In other respects
these types resemble each other so closely that we are inclined
to believe that they must be regarded as representing the same
form, but two physiological varieties, one having lost the
liquefying power which the other still retains. Similar vari-
ability is especially true in regard to pigments. Our experience
has convinced us that whereas some pigments, like brilliant
reds and deep oranges and greens, are distinctive characteristics
of groups, the pale yellow colors and the white pigments are
so sure to run into each other by intermediate variations that
they are of very little value in distinguishing types. In the
following pages it will be seen that a white and a yellow type
of coccus has been described and recognized as different groups,
but the intermediate forms which we have found convince us
that they are really practically the same thing with slight
physiological variations. Even in the question of morphology,
which is usually regarded as the most manifest criterion for
distinguishing types, something of the same appears. The
distinction between the short rods forms and the coccus forms
is by no means a sharp one. Cultures which have been de-
scribed in our own laboratory as short rods have been sent to
other laboratories and been described as coccus forms. To
eliminate the confusion arising in all of these ways is manifestly
as yet impossible, but we have endeavored in the arrangement
of groups given in the following pages to recognize the sharper
distinctions and to state, where possible, the connections of the
groups by intermediate forms which have been discovered.

In our studies, as involved in the following pages, we have
endeavored to work upon and describe only such types as. are
usually found zz zature. Cultural varieties are not generally
included in our list. ‘The work in our own laboratory and
elsewhere has shown that by modifications of the culture con-
ditions an endless series of modifications may be produced in
the descendants of the same original stock culture. The power
of producing pigment may be changed, the power of liquefying
gelatin, the shape of the bacteria themselves may be quite
modified by different culture methods. It does not fall within
our purpose to include such cultural varieties. We are inter-
ested, in this paper, not in knowing through what variations

CLASSIFICATION OF DAIRY BACTERIA. 97

different stock cultures may be forced, by modifying conditions,
but rather what are the physiological characters of the types
that actually exist under the conditions of nature and which
may be found in normal dairy products. Most of the types
described in the following pages, therefore, ave actual cultures
obtained from dairy materials, and in no case do they represent
modifications of such forms, except such modifications as natur-
ally occur under the normal cultures in the laboratory. A few
cultures sent us by others form an exception to this rule.

To our mind the aim of classification of such a group of
bacteria at the present time should be as follows:

1. To recognize the groups of bacteria in the terms above
used, and then to describe these groups in such a way that
they may be with tolerable ease recognized by others working
upon the same subject.

2. To recognize the kinds of variation possible within the
groups. By thisis meant the study of as many natural cultures
of the members of the group as possible, noting what variations
appear in the different cultures, as isolated from milk, and thus
determining to what extent the group characters are modified
in the different forms of bacteria as isolated from actual milk
products.

3. To find, if possible, the natural limits of such groups.
By this, of course, is meant to determine the limits within
which the variations may occur, and yet the type in question
may be legitimately regarded as belonging to the group in
which it is placed. If no such limits could be found, natur-
ally the whole question of classification of bacteria would be
pretty nearly hopeless. It may be that as information accumu-
lates, it will be found that all of these groups so run into each
other as to make it impossible to logically separate them. This
is, of course, what would be naturally assumed upon the
general theory of evolution of types, but whether such is the
case can only be determined after an immense amount of data
has been accumulated. At the present time, with the data at
hand, it seems to be possible to arrange these forms into fairly
distinct groups which, though connected more or less with
intermediate types, are nevertheless recognized with as much
certainty as the different types of animals and plants may be
recognized.

98 STORRS AGRICULTURAL EXPERIMENT STATION.

In determining these groups the primary question that arises,
of course, is what characters shall be used to separate the
groups. While all characteristics should, of course, be taken
into account, some are clearly of more importance than others.
There is among bacteriologists as yet no consensus of opinion
as to what characters are of most importance for this purpose.
It is, however, quite generally thought that morphological
data are primary and that these should be the first points of
distinction between these organisms. We are inclined to be-
lieve that this is true, but as intimated above, we have been
forced to think that it is necessary to make certain modifica-
tions of this statement. The cocci and bacteria certainly run
into each other in such a way that it is sometimes simply im-
possible to determine whether a culture should be called an
extremely short bacterium or a coccus type. As already stated,
the same culture will be described. by one person as one and by
another as the other of these two forms. Moreover, the classi-
fication of the spherical forms as usually accepted to-day into
the streptococcus and micrococcus groups appears to be imprac-
ticable in many cases. We have found many forms of bacteria
in which it is practically impossible to determine whether we
are dealing with a coccus that divides in one plane only or in
two planes. Moreover, in one or two cases we have clearly
found a spherical form that multiplies for a long time by divid-
ing in one plane, producing a long chain, which would be
then naturally called a streptococcus, and then the whole series
of spheres divided in the other plane, at once giving chains of
pairs. This observation has been made once or twice in our
own laboratory and has been confirmed by Winslow. Whether
to call such a type a streptococcus or a micrococcus is evidently
a problem. We have furthermore found in our study that
many forms which are described as streptococci are identical in
every character with others described as micrococci except this
one point of the method of division. These facts have con-
vinced us that this distinction of the micrococci and strepto-
cocci is an uncertain one, and we have not found it possible to
follow it out with accuracy. We have therefore used this
character as a secondary rather than a primary one in distin-
guishing types. The question of flagella present upon micro-
organisms has appeared to us to be of more importance than

CLASSIFICATION OF DAIRY BACTERIA. 99

some of the other morphological characters. We have in our
classification, therefore, sharply separated the motile from the
non-motile rods and the peritrichic from the monotrichic bacilli.
In a few of our early described species in which flagella were
not made out we have found it possible to group them from
their other characters with later isolated cultures which are
more carefully described. '

The question of the liquefaction of gelatine has also been an
open one, for the data at our command seem to suggest that
here, too, we have a character that is somewhat variable.
Some types show this property of liquefaction only after two
or three weeks’ growth, and we have considerable data to indi-
cate that the power of liquefying may be completely lost; in-
deed, among the organisms isolated from milk we have occa-
sionally found two that are identical in every respect except in
this power of liquefaction, and organisms that have such pecul-
iar character as, for instance, the power of producing a pink
fluorescence, as to convince us that we are really dealing with
the same organism, but one in which the property of liquefying
gelatine is capable of being totally lost.

In the grouping of bacteria in our key we have used prom-
inently the power of fermentation of sugars as a means of diag-
nosis. This characteristic is one which has become recognized
recently as quite significant, and for the purpose of the study
of the groups of the bacteria of milk, it is evidently one of
exceptional importance. The fermenting power of sugar is
closely related to the action of the organisms upon milk, and
clearly from the standpoint of dairy bacteriology a grouping of
bacteria with this as a basis is one of the most practically
useful methods of grouping these organisms. But here, too, it
appears to us that variations are common. Some of our groups
contain organisms which apparently show considerable differ-
ence in their power of fermenting different sugars. It is com-
mon to separate in different groups any organisms which would
ferment dextrose but not saccharose from one that would fer-
iment the two sugars. As we have compared together the large
numbers of cultures from various sources that we have made,
we have found such wide variations in this power of ferment-
ing sugar that we have been inclined to believe that here,
too, we have a variable factor, and that whereas the general

100 STORRS AGRICULTURAL EXPERIMENT STATION.

power of fermenting sugar may be taken as a valuable and
important criterion in separating groups, there may be vari-
ations among the members of this group in the kind of sugars
they can ferment and the readiness with which they can carry
on this action. At all events we believe that the variations in
the power of fermenting sugar are only of value in separating
closely allied varieties from each other, but quite insufficient
to characterize different groups.

The production of pigment by bacteria is one of the charac-
ters frequently more striking than any other. The question of
the value of this characteristic in separating organisms from
each other has been much discussed. It has been many times
shown that this power of producing pigment is capable of
variation. It is certainly true that some organisms that are
able to produce pigment under some conditions lose this power
after cultivation. 2. rudensis, for example, which, when first
isolated, produced a brilliant red pigment, wholly lost this
power in cultivation, and when the organism had reached my
laboratory many months from its original source, it was a pure
white Bacterium, showing all the other characteristics of the
genus, but having lost its pigment-producing power. If the
power of producing pigment is thus capable of variation, it
indicates, of course, that it is not a criterion to separate types
radically from each other. Nevertheless, in the general study
of micro-organisms this is one of the characteristics which must
be taken into consideration. Even though we recognize that
some pigment-producing organisms may, under conditions of
long cultivation in the laboratory, be converted into non-pig-
ment-producing types, it is none the less important for us to
recognize the pigment-production as a distinct characteristic of
the organisms as found in milk productions.

For practical purposes it is a matter of less importance to
know that a red pigment may cease to be developed after long
cultivation in the laboratory than it is to recognize as a distinct
type of dairy organisms a bacillus producing a red pigment.
We have therefore used the pigment production as one of the
characteristics for separating our groups. From our observa-
tions we have been inclined to think that red, orange, brown,
and green pigments are commonly distinctively characteristic,
that a lemon yellow color is also a character which remains

CLASSIFICATION OF DAIRY BACTERIA. IOI

tolerably constant, but paler yellows are of far less significance.
In our classification the color of yellow or yellowish cannot,
therefore, be .regarded as particularly important, and we are
convinced that many of the varieties described as yellow are
identical with others described as white. Where we have good
reasons for believing this is the case, we have mentioned the
fact in the following pages by cross references.

In our opinion the other characteristics commonly used for
describing bacteria are less significant. ‘The appearance of the
colony in gelatine is subject to wide variations, and while some-
times it is useful, it cannot be relied upon as a rule as of very
great importance. ‘The growth on potato varies more or less
widely with the nature of the potato, and so decided are these
variations that we have ceased to place much confidence upon
the characteristics here described. The growthin bouillon has
appeared to us to be of less importance than some have thought,
even the formation of a scum being somewhat variable in differ-
ent cultures of the same organism.

The question of nomenclature is, of course, a puzzling one in
dealing with a new group of organisms, whose inter-relations
are as problematical as that among bacteria. Nevertheless, it
is desirable that certain names should be given in our classifica-
tion. In the following pages, we have adopted the following
plan. We have named each of the general groups of organisms
which we have recognized as more or less clearly marked types.
‘The names that we have given to these are in all cases where
possible names already applied to the chief members of the group.
In many cases we cannot find among described bacteria any which
agree closely enough to admit of identification, and in regard to
many others, while there are among described types some which
agree with ours as far as they go, the published descriptions are
too incomplete to admit identification. In both of these cases
we have given new names. Inour names we have in practically
all cases used the word /actis in thename. Weare fully aware
of the objections to using three names for a species, but in these
cases the manifest advantage of indicating by the name the fact
that the organisms are associated with and found in milk prod-
ucts is so great that we regard it as sufficient to overcome the
disadvantage. We have used this method in all cases except a
few well known organisms where it was manifestly unwise.

102 STORRS AGRICULTURAL EXPERIMENT STATION.

Underneath these groups we have tried to recognize the
types of varieties which we have found and which are there-
fore liable to be isolated from any samples of milk, and for the
present we have grouped these as varieties A, B, C, D, etc.,
under the general name of the group. In this way we recog-
nize the general classification of bacteria, and at the same time
obtain an impression of the variations that develop within the
types themselves.

Method of Description.—In describing a long list of species of
bacteria such as are included in the following pages, there are
two different purposes to be considered, each of which would
involve a different method of procedure. First, there is the
question of dzagnoszs, which would call for such an arrange-
ment of important characters as will enable other bacteriologists
to identify forms which they may have in hand with those
already described. There is, second, the need of such a com-
plete description of the organisms in question, with a// its other
characters as will finally and definitely describe them for a
permanent record. In accordance with which of these two
purposes we have in mind the method of description would be
varied.

In our own opinion at the present time the first of these two
_.points is the more important. Until we know more about the
general relations of bacteria, it is more important to have such
a grouping of the types as will enable others to identify the
cultures which they may have under observation with those
already described. The more minute detailed description of
species will undoubtedly of necessity come later. But when
we recognize that such considerable variations of the same type
are possible, and certainly do occur under cultural conditions,
it becomes evident that the minute description of the physio-
logical characters of the different cultures becomes of compara-
tively little value. If the types are not constant in all of these
points, it is certainly of no great value, at least in the present
state of bacteriological science, to describe the particular char-
acteristics of a culture at any particular time, especially if it is
true that when cultivated in the laboratory for six months,
these characters may largely change. Hence we have con-
cluded that with the present state of bacteriological science the
more valuable form of classification will be one which will

CLASSIFICATION OF DAIRY BACTERIA. 103

select the salient features, and so far as possible those that
are the least subject to variation, and so tabulate them as to
make it feasible and convenient for other students to recognize
the types in question and to identify them with any other form
they may be studying. We have, therefore, in the following
pages placed emphasis wholly upon the question of drief diag-
nostic characters and in arranging them in such a form that
they can be easily utilized, rather than in a detailed descrip-
tion of species.

We have also learned from experience that unless the char-
acteristics of species can: be clearly and distinctly tabulated, it
is almost a hopeless task for anyone to identify a new culture
with one previously described. The methods of description
that have been in vogue in the first decade of the study of
bacteria have been to describe in somewhat verbose detail all
sorts of minute characteristics, which later discovery has shown
to be of little or no significance. These have been commonly
arranged in no order and have never been tabulated. The
result has been that it is almost impossible to identify any
previously described type of bacteria. Consequently the bac-
teriological literature has seen a constant succession of new
types described, and when these descriptions are compared
with each other it becomes more and more evident that the,
same general type of bacterium has been described over and
over again by different bacteriologists and given name after
name. Undoubtedly some of our most common dairy organ-
isms have been given a dozen or fifteen names, due to this un-
fortunate method of describing species by long details. We
have endeavored, so far as possible, to correct this error in our
classification, by excluding all unimportant data and keeping
only what seemed to be the distinctive and more or less con-
stant characters, and then to arrange these in such tabular
form by methods already known so as to make it possible to
use them conveniently.

We have arranged our classification as follows: We have
first inserted a detailed description of the different bacteria which
we recognize under different names. Instead of attempting
however, to describe these to any great length, as would be
necessary if all of the characters were filled out which are de-
manded by the description blanks of the American Society of

104 STORRS AGRICULTURAL EXPERIMENT STATION.

Bacteriologists, we have endeavored to insert only the salient
characters. It has appeared to us that the numerous details
only confuse one in an attempt to identify and classify bacteria.
When it is so evident that these physiological characters, which
are largely the basis pf these long descriptions, are capable of
such great modification by culture, when it is evident that the
cultures of bacteria which one obtains are likely to come from
a great variety of sources and therefore to have been subject to
a large variety of different conditions, it is evident that we may
expect an almost endless variation in the physiological charac-
ters of the different cultures. If we donot identify two cultures
of bacteria as the same until we find that they agree exactly
in all of the long series of characters given in the society
blanks, we shall practically never identify any two types of bac-
teria as thesame. It has seemed to us that this is an absurdity
and that the recognition of the relations of bacteria will be
advanced by omitting insignificant details, concentrating our
attention upon the more important characters. For this reason
we have in these descriptions omitted much of detail and have
only given such points of description as have seemed to be
most salient.

Following the detailed descriptions we have given analytical
keys and tables covering all the types. We recognize the
fact that the bacteria of milk may be grouped in three gen-
eral groups, the Coccaceae, the Bacteriaciae, and Bacilliariciae.
These groups are morphological ones, and while, as has already
been pointed out, we are convinced that to a certain extent
they run into each other and cannot be sharply distinguished,
they do represent fairly well marked groups. Under each
of these heads we have given an analytical key, in which
the important characters are used for purposes of diagnosis.
By means of this key it is possible to trace readily to its
group or to its allies any type of bacterium of which we
have a tolerably complete description. These analytical keys
are given for each of the great groups of bacteria. Follow-
ing these analytical keys we have given finally a tabulated
key of all of the important characters of the different species.
In the preparation of this key we have made use of the plan
adopted by the American Society of Bacteriologists, selecting
the important characters which can be indicated by positive

CLASSIFICATION OF DAIRY BACTERIA. 105

and negative signs. This plan, advanced some years ago by
Fuller, has proved useful, and inasmuch as it has been pro-
visionally adopted by the American Society, we have made use
of it in the tables of our key. In our own opinion the plan
advanced by Gage and Phelps in using numbers that have cer-
tain arbitrary meanings is far more satisfactory and far more
usable than this plan which the American Society has adopted;
but it is better to have uniformity in the matter even at the
expense of some loss, and we have therefore adopted the plan
of the American Society. All of our species are arranged in
tables in this way, the tables being so grouped that organisms
which are most closely related to each other come in proximity.
With this plan it is only necessary in order to identify the rela-
tions of a new type of bacterium, to fill out on one of the cards
‘prepared by the American Society, the blanks left for the in-
sertion of the characters, and then to place this blank upon the
table in the following pages and running it up and down the
page until there is found one organism with which the plus
and minus signs practically agree. When this is found it is
sure that the allies of the species under consideration have been
identified, though it may not agree in all details. In this table
we have also used the group numbers as directed by the Ameri-
can Society of Bacteriologists, this group number being another
important aid in identifying a culture and placing it among its
allies.

In a number of cases we have found that what.we believed
to be the same organism comes under two different heads in
our classification; for instance, a culture has been described as
a Bacterium and another one as a Coccus, and yet when care-
fully studied out in all other respects they are found to be the
same. Under these circumstances we have been uncertain as
to where it should be classified and we have therefore placed it
under both divisions of Coccacece and Bacteriaciae. The same
thing has occurred occasionally with organisms that liquefy or
do not liquefy gelatine. In all such cases cross references are
inserted to indicate these probable relationships.

It should be finally stated that the forms which we recognize
in the following pages which we name must be regarded as
groups and not species. This is not at all material, inasmuch
as we have no conception whether the term species has any

x

106 STORRS AGRICULTURAL EXPERIMENT STATION.

meaning whatsoever among bacteria. Our names, therefore,
refer to groups under which there are in some cases many
sub-varieties, in other cases few and in some cases uo sub-
varieties. From the standpoint of practical dairying the sub-
varieties are sometimes of more significance than the general
type. For example, under the head of Bacterium lactis acidi
there are certainly a number of varieties differing in re-
spects which are of extreme importance to the dairyman.
They differ in the amount of acid they produce and in their
power of curdling milk. So, too, under the head of Bacterium
lactis aerogenes we have clearly a group of organisms differing
in many important characters. Among these characters there
is wide difference in the extent of the gas production; some
cultures producing only a small amount of gas, others produc-
ing gas in prodigious quantities. While these variations in the
amount of gas cannot, in our opinion, be regarded as points by
which the group should be separated from each other in a
scheme of classification, they are of the highest importance to
the dairyman. The variety which produces a small amount of
gas would be consistent with the best dairy products, while the
presence of the other variety would totally ruin a lot of butter
and more surely ruin a lot of cheese. These sub-varieties, in
short, are of extreme importance to dairymen, and the careful
study of these varieties is a problem which should be con-
sidered carefully in the future; but in a general classification of
bacteria it is, in our opinion, at present quite impossible to rec-
ognize all of these types by name, and our present plan is,
therefore, only to group them as varieties under the general
group name.

METHOD OF STUDY AND DESCRIPTION.

A few words are needed as to the method of obtaining the
data tabulated in our descriptions. The zso/atzon of the bacteria
is usually accomplished by the use of litmus gelatine. The pre-
paration of this litmus gelatine has been described in a former
publication. (Bacteria in Milk and its Products, P. Blakistons
Sons.) This has been used because our experience has shown
that it gives a better differentiation of colonies than other solid
media. After isolation the cultures are purified by the ordinary

'

CLASSIFICATION OF DAIRY BACTERIA. 107

method of replating, and, after purification, they are inoculated,
upon agar streaks. After about two days’ growth on agar they
are inoculated into the various other culture media.

The morphology of the various organisms was determined from
fresh agar streak cultures although, to determine the formation
of spores, it was sometimes necessary to use older cultures. To
determine the motility we have usually used an ordinary bouillon
culture of 12-24 hours’ growth. We have found that a careful
study of a hanging drop of such a bouillon culture is most satis-
factory for this purpose. To determine the presence of flagella
we proceeded as follows: from an actively growing bouillon cul-
ture a drop was removed with a platinum loop and spread thinly
over the surface of solidified agar. This was then incubated at
37° for 12 hours. A-small quantity was then removed, diluted
in three successive drops of sterilize water and stained by the
well known Loeffler method.

‘The culture media we have used have been those ordinarily
employed. Our bouillon, gelatine and agar have been made with
Liebig’s beef extract instead of chopped beef, because of greater
convenience and uniformity. The fermentation tube test was
made with dextrose, lactose and saccharose, 1 per cent. of these
sugars being added to ordinary bouillon. In all the cultures
tested by us in the last five years we have used all three sugars.
The mz/k which we have used has in all cases been skimmed
milk, sterilized by boiling for 10-15 minutes on three successive
days. Potato cultures have been made by cutting plugs from
large clean potatoes, slicing them once obliquely, and then soak-
ing them in running water over night. After this they were
placed in tubes and sterilized in an autoclave.

In general the terms which we have used in our descriptions
have been those suggested by Chester and adopted by The
American Society of Bacteriologists. In stating whether or not
an organism produces acidity we refer to the production of acidity
in dextrose bouillon. In some cases, as will be seen, organisms
produce acidity in lactose but not in dextrose; but these are rare.
The detection of acidity has been made by meaus of carefully
prepared litmus paper. This paper we have prepared ourselves
from neutral litmus. The test for acidity has been made in
about two days, and also later. This method has been adopted,
both for the fermentation tubes and for milk. In determining

108 STORRS AGRICULTURAL EXPERIMENT STATION.

the action upon milk we have always used both the temperature ,
of 20° and 37°. In our descriptions in the following pages we
have not referred to the action of these two temperatures, except
in cases where it was different at the two temperatures. It will
be understood, then, that the actions given as taking place in
milk occur at both temperatures unless otherwise specified, al-
though usually, of course, more rapidly at the higher temper-
ature. The digestion of milk has been determined by eye only.

We have hitherto done no work upon strictly anaerobic bac-
teria. Most of the forms which we have described have been
aerobes, although some grow more readily if they do not have
too abundant a supply of oxygen. ‘The question of their rela-
tion to oxygen we have determined from their surface growth
on various media, their growth in the closed arms of the fer-
mentation tubes, and also from their growth under a mica plate
which we have always made and frequently found useful, al-
though we have not tabulated it in the following pages.

DETAILED DESCRIPTION OF TYPES.

A. NON-LIQUEFYING COCCI.—STREPTOCOCCI AND MICROCOCCI.
I. Types that do not produce acid.

MM. lactis rosaceus n.s. A pink Micrococcus. This organism, orginally
found in 1903, has been found twice since, in milk, in this vicinity. All cul-
tures agree in essential particulars.

Morphology.—Size, .8uin diameter. Stains by the Gram method.

Gelatine colony.—Surface colony reaches the size of I mm., with a nucleus
and a light outer zone. he color is pink. On “tmus gelatine it produces a
bluish colony which is not acid.

Gelatine stab.— A needle growth and a spreading pink surface,

Agar streak.—A luxuriant pink growth.

Fermentation tubes.—No acid nor gas is produced in any sugar bouillon, and
no growth in closed arm.

Bouillon.—A sediment and a slight turbidity, but no pellicle.

Mitk.—Rendered slightly acid and shows a slight pinkish sediment. It
becomes somewhat slimy.

Potato.—A very luxuriant, thick, moist growth of a pink color.

Grows at both 20° and 37°. Aerobic.

M, lactis citreus Bon. s. A yellow, non-acid Coccus. (Perhaps= J, citri-
nus Migula). The similarity of this culture to our yellow liquefying coccus leads
us to think that they are perhaps the same and hence the use of the same name.
(See AL. lactis citreus A, p. 118.) This organism has been found but once in
milk in Middletown. Its characters are as follows:

Mor phology.—Size, .84. No chains produced. Stains by the Gram method.

~

CLASSIFICATION OF DAIRY BACTERIA. 10g

Gelatine colony.—A distinctly round, thin, yellow surface colony, which be-
comes about I mm. in size. Not characteristic.

Gelatine stab.—A good needle growth and a moderately luxuriant surface
growth, of a yellow color.

Agar streak.—A luxuriant, yellow growth.

Fermentation tubes.—There is no production of acid or gas in any sugar
bouillon, and no growth in closed arm.

Bouwillon.zAn abundant sediment and turbidity, and a thin pellicle on the
surface.

JTilk,—At both 20° and 37° the milk is rendered acid, but it is not curdled
or otherwise affected. ‘The acidity is very slight.

Potato,—An abundant, canary-yellow growth, and the potato is discolored.

Grows well at both 20° and 37° ~— Aerobic.

M, lactis flavus n.». An orange, non-acid Micrococcus,

Morphology.—A micrococcus. Size, .5¢-.84. Stains by the Gram method.

Gelatine colony.—A round, smooth, thick, homogeneous, orange-colored
surface colony.

Gelatine stab.—A good needle and surface growth, with an orange color.

Agar streak.—Luxuriant, moist, smooth, of an orange to a red-brown color.

Fermentation tubes.—No acidity or gas in any sugar bouillon. Dextrose and
lactose may be rendered alkaline. No growth in the closed arm.

Bouillon.—A sediment, a turbidity, and a pellicle; or the turbidity and pellicle
may be wanting, the latter characteristic belonging to a second culture isolated
at a different time.

Milk.—Is rendered acid both at 20° and 37°, but no other change is pro-
duced, except a slight yellow color in some cases.

Potato,— Moderate to luxuriant, moist, smooth, red-brown to orange.

Grows at 20° and 37°. Aerobic.

We have found this organism several times. ‘he different cultures differ,
however, in small points. The chief lines of difference were the following: In
color it ranges from orange to bright yellow. In two cultures the gelatine
colony was white rather than yellow, and in these same two cultures milk was
not rendered acid. One culture curled milk, acid. One culture grew in the
closed arm of the fermentation tube, while the others did not. Weare not in-
clined to think these differences sufficient to separate them as varieties.

Apparently this is identical with AZicrococcus D. of Barthel, and perhaps
with M7, aurantiacus Cohn.

M. lactis viscosus Cn. s. A slimy milk Micrococcus. This culture was
sent us by Harrison and was isolated by him from Pasteurized milk. We have
not found it ourselves. Its failure to make milk acid and its dark colored
growth on potato seem to distinguish it from any other slimy milk micrococci.
Hence we call it variety B. (See p. 114). The characters are as follows:

Morphology.—A micrococcus. Size, .84-.gu. Gram stain positive.

Gelatine colony.—A thick, round, smooth, white colony. In old cultures the
gelatine is turned green. On /éémus gelatine the colony is coarse, granular and
nucleated. The surface growth is rather transparent. Below the surface it is
opaque and brownish.

Ilo STORRS AGRICULTURAL EXPERIMENT STATION.

Gelatine stab.— A vigorous needle and surface growth.

A gar streak.—Moderately thin, spreading, white.

Fermentation tubes.—Neither of the sugar bouillons is rendered acid, nor is
there any gas produced, nor growth in the closed arm.

Bouillon.—A sediment and turbidity, but no pellicle.

Milk,—No change except the production of a decided sliminess.

Potato,—A luxuriant, thick growth of a slaty-gray color, turning to blue or
black, and old cultures to an olive-green. The potato itself is discolored.

It grows well at both 20° and 37° ~— Aerobic.

The organisms that produce slimy milk are numerous. They do not by any
means form a class by themselves, for this property of rendering milk slimy
seems to be found scattered widely among bacteria. Apparently the type that
most commonly. produces trouble in dairies is a Bacterium. The following
organisms have been described as causing sliminess, some of which are included
in our list and others omitted as insufficiently described: AZ. viscosus Bechamp,
Actinobacter du lait visgueux Duclaux, Actinobacter polymorphus Duclaux, 8B.
mesentericus vulgatus Fligge, M. mucilaginosus Migula Migula, B. lactis
pituitost Loeffler, Strep. Hollandicus Weigmann, Bact. lactis longi Troili-Petter-
son, B. Guillebeau c, Freudenreich, Bact. Hessii Guillebeau, W/. Freudenreichti
Guillebeau, Carpho. coccus pitiutoparus Hohl, Coccus lactis viscosi Griiber, 2.
lacto rubifaciens Griiber, Bact, lactis acidt Leichmann, Bact. aerogenes Escherich,
B. Harrisonit, M. viscosus A, and some others. This list includes almost every
type of bacteria, so that the characteristic of slimy milk cannot be taken as
forming a group mark.

M. lactis arborescens,—This was described in our former report and has been
found twice since.

Morphology.—A micrococcus. Size, .74 in diameter.

Gelatine colony.—A myceloid colony about I mm. in diameter. Sometimes it
is smooth.

Gelatine stab.—An arborescent needle growth anda surface growth,

Agar streak.—A luxuriant, white, smooth moist growth.

Fermentation tubes.—Not determined, but doubtless neither acidity nor gas.

Bouillon.—A turbidity, sediment and a tenacious scum.

Milk.—No action or a very slight alkalinity.

Potato.—A spreading, brownish growth, not very luxuriant.

Grows at 20° and 37°. Aerobic.

Galactococcus versicolor Lux. A white non-acid coccus. One of the most
common types of milk organisms proves to be a small white coccus that does
not produce acid and without any very marked characters, There are a large
number of these which, although showing some variations, are so much alike
that we regard them as forming a single type. They are extremely common in
milk and are certainly found in the udder very commonly. We have adopted
the name first given by Lux as fairly distinctive, although we think it would be
better to use the name Streptococcus versicolor. We can recognize two types
although no sharp line can be drawn between-them.

Morphology.—A streptococcus. Size, .54-1.5u. Never more than three or
four joined together. It commonly stains by the Gram method.

CLASSIFICATION OF DAIRY BACTERIA. III

Gelatine colony.—Spreads slightly over the surface, white, yellowish or
brownish, lobed or round or rather thin. The colony is not characteristic.

Gelatine stab,—An abundant needle and a surface growth.

Agar streak,—White to yellowish, moist, not luxuriant.

Fermentation tubes.—No acidity in any sugar bouillon; there is no gas or
closed arm growth.

Bouillon.—A sediment and turbidity but no pellicle.

Milk. —Usually no action, although a few cultures show a slight acidity.

Potato.—Very scanty, grey-white, or sometimes no growth. ~

Grows at both 20° and 37°. Aerobic.

Variations from the above are shown in color, from a white to a yellowish; in

the formulation of a pellicle in one culture; and in the faiiure to stain by the
Gram method.

Variety A.—We recognize as a distinct variety one form that grows luxu-
riantly on potato and has a less tendency to form a yellowish pigment. This
type has also a slight tendency to produce an acidity in milk, in this respect
agreeing with A7. candius of Barthel, which is probably the same, Variety A
is equally common with the type described.

One culture which would naturally belong here shows relations to JZ. lactis
albus in that it digests milk, but does not liquefy gelatine. Its characters are
as follows:

Variety B. Digesting milk without liquefying gelatine.

Morphology.—A micrococcus. Size, .6%. The Gram stain is negative.

Gelatine colony.—Very small, yellowish, surface colonies, smooth, convex,
entire, moist.

Gelatine stab.—A needle growth and a convex surface.

Agar streak.—Filiform, raised, smooth, transparent, moist, luxuriant. In
some cases wrinkled.

Fermentation tubes.—No acidity or gas in any bouillon. Usually no closed
arm growth, but one culture showed growth in closed arm.

Bouillon.—A sediment and turbidity; a pellicle sometimes forms and some-
times not.

Milk.—Becomes alkaline'and digests. This digestion becomes complete in
three weeks, and is unusual since the organism does not liquefy gelatine.

Potato.—Luxuriant, filiform, capitate, cream color. Potato not discolored.

Grows at 20° and 37°. Aerobic.

The description of this Coccus appeared Yo us to agree closely with that of
Streptococcus pyogenes. To see how closely they agree, we obtained a culture
of the latter from the bacteriological laboratory at Yale Medical School, and
compared it with our cultures side by side. In all cultural characters we found
that the two agreed so closely as to be indistinguishable. Whether this indicates
that they are identical we would not at present determine, but the identity in
cultural characters of this common pathogenic Coccus and this most common
dairy organism is at least suggestive. In fresh milk this type is probably the
most common of all dairy bacteria.

I12 STORRS AGRICULTURAL EXPERIMENT STATION.

Il. Types that produce acid in dextrose or other sugars.

S. lactis fuluus n. s. A brownish-red Streptococcus. This was obtained
once directly from the udder and once from city milk. Another culture was
obtained from stable dust which agree in all points except those indicated in
brackets.

Morphology.—A streptococcus. Size, .7u. Gram stain is positive.

Gelatine colony.—Small and dense, 5 mm. in diameter, rather thick, round,
white. The colony in litmus gelatine is acid. [Brownish-pink to orange. ]

Gelatine stab,— A needle growth and a thin surface growth.

Agar streak,—Luxuriant, thick, moist, translucent, reddish-brown [orange-
yellow].

Fermentation tubes—All three sugar bouillons rendered acid; there is no gas

vor closed arm growth.

Bouillon.—A sediment, turbidity, but no pellicle. [No turbidity.]

Milk,—Is acid and curdles.

Potato.—Luxuriant, brown.

Grows at 20° and 37° — Aerobic.

One culture obtained from New York milk shows some rather striking charac-
ters, and we recognize it as a separate variety as follows:

Variety A.—This agrees with the above except in the following: Size,
1.2u-1.4@. Gram stain negative. Gelatine colony ringed and irregular, show-
ing acid in litmus gelatine. Bouillon with a pellicle. Milk, curdled acid at 37°
only. Potato has luxuriant growth and is discolored.

M. lactis aureus n.s. Many different cultures of yellow, acid Cocct have
been found in this vicinity, from Canada and New York. They are found in
milk, in cheese, in butter, and in stable dust. Although they do not liquefy
gelatine, we are confident that they are to be looked upon as non-liquefying
forms of Staph. pyogenes aureus. (See p. 121.) The general description is as
follows:

Morphology.—A micrococcus. Size, .54-1.24. Gram stain positive.

Gelatine colony.—Round, moderately thick, smooth, translucent colonies, of
a lemon-yellow color, Litmus yelatine is acid.

Gelatine stab.—A needle growth and a yellow surface growth.

Agar streak,—Luxuriant, moderately thick, smooth, translucent, yellow.

Fermentation tubes.—All sugar bouillons are rendered acid, but there is no
gas, Occasionaly there is growth in the closed arm.

Bouillon.—A sediment and turbidity, and rarely pellicle.

Milk.—Rendered acid and usually does not curdle. Some cultures, however,
curdle milk after several days at 37° Milk may be rendered yellowish.

Potato.— Usually a moderately thick, yellow growth, with the potato dis-
colored.

Grows better at 37° than at 20°. Facultative anaerobic.

Among the many cultures which we have studied, variations are found in the
following points: Size, from .5u-1.2". Litmus colony is sometimes acid and
sometimes not. Color varies from lemon-yellow to pale-yellow. In a few
cases there has been closed arm growths in the dextrose, and sometimes in lac-
‘tose. In two cultures a pellicle formed on bouillon. Milk always acid but

CLASSIFICATION OF DAIRY BACTERIA. 113

only about half of our cultures curdle it. Potato varies from scanty or no growth
to luxuriant. We do not regard these variations as sufficient to warrant us in
distinguishing them under different names.

S. lactis aureus n. s. An orange red Streptococcus. This organism is
apparently sufficiently different from the last to be separately named. It was
found in a Camembert cheese sent from France. Several varieties occurred all
together in the same cheese and they are, doubtless, physiological varieties of
the same organism.

Morphology.—A streptococcus. Size, 1m-1.24. Gram stain positive.

Gelatine colony.—A round, thick, rough, opaque, surface colony, of a creamy-
white color. On litmus gelatine it is dense and not acid, with an irregular
edge. One culture was thin. ?

Gelatine stab.—A needle growth and a raised surface growth; orange.

Ayar streak.—A luxuriant, rough, orange-yellowish to greenish-yellowish
color; sometimes dull and wrinkled.

Fermentation tubes.—All sugar solutions are rendered acid, but there is no
closed arm growth nor gas.

Bouillon.—A sediment and a turbidity are produced, and, after several days,
a pellicle.

Milk.—After some days rendered acid and curdled. Some of the cultures
did not produce acid and did not curdle; a slightly sour odor.

Potato.—A thick, rough, opaque, yellow, luxuriant growth; potato may be
discolored.

Grows at 20° and 37°. Aerobic.

We have found several white coccus forms that produce siimy milk. The
differences between them are considerable, although it may be that they are all
variations of the same type. At present we recognize three.

S. lactis viscosus n. Ss.

Morphology.—A streptococcus. Size, .84-.94. The Gram stain is negative.

Gelatine colony.—A shiny, pale-yellow, round or lobate colony, 1 to 2 mm, in
diameter. It is commonly viscous.

Gelatine stab,—A needle growth and a surface growth, producing a nail
culture,

Agar streak.—An irregular lobate surface growth, quite luxuriant, viscous.

Fermentation tubes.—All three sugar bouillons become acid and there is
growth in the closed arm, but no gas is produced.

Bouillon.—A sediment and a turbidity, and also a pellicle.

Milk.—Rendered acid and curdled after three or four days. The milk
becomes very slimy.

Potato.—A luxuriant, dull, paste-like growth, of a gray to a yellowish color.

Grows at both 20° and 37° =‘ Facultative anaerobic,

This description is from the slimy milk organism of group V. as given by
Harrison. We have found an organism at Storrs which is apparently the same,
and which we had previously named MM, /actis viscosus. We will call this a
distinct variety, and it differs from the above described in the following points:

Variety A.—The agar growth is scanty and not viscous. No pellicle is
formed on bouillon, Growth on potato is luxuriant and the potato is discolored.

ita STORRS AGRICULTURAL EXPERIMENT STATION.

To this same group apparently belongs M7. Freudenreichii, as well as can be
determined by the incomplete descriptions given.

Reference must also be made to MM, Jactis viscosus B., differing from either
of those described, so as to be located elsewhere in our scheme. (See p. 109.)

The white cocci which produce acid and no slime are very numerous and their
separation into groups is uncertain. We have tried to divide them into
several types but the divisions between them are not satisfactory. Each of the
groups shows variations, and it may be better to unite them altogether. For
the present, however, we recognize the following divisions, based primarily
upon their action on sugar and the abundance of their surface growth.

S. lacticus Kruse. This includes the smaller acid cocci with a scanty surface
growth. This type is very closely related to, if not identical with, Bact. lactis
acidi, The organism that commonly causes the souring of milk shows some
variation in its morphological structure. It is sometimes clearly a short coccus
form, while in other cases it is decidedly more elongated. In cultures, too,
similar variations have been found. It commonly appears as a short rod,
clearly longer than broad, and as such has been usually described as a Bac-
terium and named Bact. dactis acidi by Leichmann. By this name it is commonly
referred to in literature. But cultures of Bac¢. dactis acidi when grown in lactose
bouillon frequently, at least, appear as streptococci, and Kruse (Cent. f. Bac.
u. Par. I., XXXIV., p. 737, 1903), and Heinemann (Cent. f. Bac. u. Par. II.,
XVI,, p. 538, 1906), have recently insisted that there is no such thing as Bact.
lactis acidi, all of the organisms which have been so named being really cocci,
which should be called S. /acticus. Upon this point we will at present express
no opinion beyond the statement that the organisms as found in souring milk
are sometimes streptococci and sometimes clearly longer than broad, and hence
would be naturally classed as Bacterium. We regard it better at present, there-
fore, to retain the name Bact. /actis acidi for the milk type as it commonly ap-
pears, and to recognize the streptococcus form independently under the name
given by Kraus It has the following characters:

Morphology.—A streptococcus with commonly short, but sometimes long,
chains. Size, .5u-1". Gram stain positive.

Gelatine colony.—Extremely minute, white colonies, occasionally slightly
yellowish, rough and dense. In ditmus gelatine the colonies are always acid.
The surface growth is always slight and usually absent entirely, and it grows
better under a mica plate. Sometimes the colonies in litmus gelatine show
minute spines on their edge.

Gelatine stab,— A moderate needle growth but no surface.

Agar streak.—The growth is hardly visible, but when it occurs is the faintest
transparent film.

Fermentation tubes.—Al\ three types of sugar are rendered acid and there is
commonly a growth in the closed arm. No gas is produced.

Bouillon.—Growth is extremely slight and sometimes invisible. A slight
sediment and turbidity may be produced.

Milk.—Rendered acid and curdled promptly, producing a typical, smooth,
acid curd.

CLASSIFICATION OF DAIRY BACTERIA. i

Potato.—The growth is usually invisible, but sometimes a scanty surface film
may be seen.

This streptococcus is very common and in some specimens of soured milk
comprises ninety-nine per cent. of all the Bacteria present. It has been found
frequently in the milk in Middletown, in storrs,in New York City, and else-
where. Where it occurs it is commonly the cause of the souring of milk, but it
is not so common as the elongated form which we have classified as Bact. lactis
acid.

The white cocci that produce acéd but no slime are very numerous and their
separation into groups is uncertain. We have divided them into six groups,
but the distinctions between them are not satisfactory. Each of the groups has
slight variations and it may be that some of them would be better united
together. For the present, however, we recognize the following, dividing them
first according to their action on various sugars, and secondly by the abundance
of surface growth produced on media.

S. lacticus J. The essential character of this type is the production of acidity
in dextrose but not in other sugars.

Morphology.—A streptococcus in pairs or short chains. Size, .6u-1.24. The
Gram stain is positive.

Gelatine colony.—This is not characteristic. It is round, white to yellowish
and spreads over the surface, forming a colony I mm. in diameter.

Gelatine stab.—A needle growth and a surface growth.

Agar streak.—Moderately abundant, white.

Fermentation tubes.—Dextrose bouillon is rendered acid, but no gas is pro-
duced and no acidity in other bouillons.

Bouillon.—A sediment and turbidity and sometimes a pellicle.

Milk.—Becomes acid, but is not curdled, and no other change is noticed.

Potato.—Growth very slight or wanting.

Four cultures of this group have been studied—two derived from milk fresh
from the udder, one from the dust of the stable, and one from cheese. They
differ slightly, as follows:

Variety A.—This culture, from the udder, produces no turbidity, but a slight
pellicle in bouillon.

Variety B.—From the udder. Produces turbidity and no pellicle in bouillon
and does not make milk acid.

Variety C.—From stable dust. Has a negative Gram stain, a turbidity and
a pellicle in bouillon, and milk is not acid.

Variety D.—¥rom cheese. Shows a turbidity and a pellicle in bouillon;
milk is acid, and there is a luxuriant potato growth.

S. lacticus [/,—Vhis produces acid in lactose and saccharose but not in
dextrose. Its other points of difference are as follows: Gram stain is nega-
tive; the gelatine colony is very small and transparent; no pellicle is ever
formed on bouillon; there is no action on milk and there is an abundant potato
growth.

116 STORRS AGRICULTURAL EXPERIMENT STATION.

S. lacticus I77,—This shows a pellicle on bouillon; the milk is usually acid
and occasionally curdled, but in some cases shows no trace of acid to litmus
paper. The potato growth is usually scanty or wanting, though sometimes
more luxuriant.

M. lactis acidi.—This name includes the smaller acid-producing cocci that
have a luxuriant surface growth on various media. Thete are very many of
them. Probably JZ. candicans of Fliigge belongs here. Their general charac-
ters are as follows:

Morphology.—A micrococcus. Size, .5#-1.24. Gram stain is positive.

Gelatine colony.—Not characteristic. The colonies are round, thin, smooth,
rather opaque and white in color. On ¢tmus gelatine no acid is produced.

Gelatine stab,— Needle growth and surface growth.

Agar streak.—A moderate, white, smooth growth, which is sometimes rough.

Fermentation tubes.—All fermentation tubes show an acid production, but no
gas and no growth in closed arm.

Bouillon.—A sediment and turbidity, but no pellicle.

Milk.—Sometimes acid and sometimes not acid. Usually not curdled,
though some cultures curdle the milk by acid production. Occasionally a
slightly sour odor.

Potato.—The growth is usually scanty and sometimes absent. It is whitish;
not characteristic.

Grows at 20° and 37°. Aerobic.

This is the most common of the milk cocci. It is found almost constantly in
common fresh milk, both here and elsewhere, where we have had an oppor-
tunity of studying it. It is clearly very similar to S. lactis acidi [/7., the primary
difference being in the amount of surface growth. We have studied very many
different cultures of this type and naturally found many variations. The most
noticeable points of variation are as follows:

Size, from .54-1.24. Gelatine stab in two cases showed no surface growth;
colony on gelatine varies much in thickness and is sometimes slightly yellowish;
the litmus gelatine colony is occasionally acid; the growth on agar varies much
in abundance, but is always more than in the lactic type.

M. lactis gigas n. s.

Morphology.—A very large coccus. Size, 1.5#. Gram stain is positive, No
chains.

Gelatine colony.—A round, thick, smooth, homogeneous, entire translucent,
cream-white colony. c

Gelatine stab.—A needle growth is produced but no surface.

Agar streak.—Growth is scanty, beaded, translucent and white.

Fermentation tubes.—All three sugar bouillons are rendered acid but there is
no growth in the closed arm and no gas.

Bouillon.—A sediment but no turbidity and no pellicle.

Mitk.—Becomes very slightly acid, sufficiently to curdle the milk in about
four weeks.

Potato.—No growth.

Grows at 20° and 37°. Aerobic or facultative anaerobic.

This coccus is characterized by its very large size and has been found several
times by us in Middletown and Storrs. It was first isolated in 1896.

CLASSIFICATION OF DAIRY BACTERIA. 117

B. LIQUEFYING COCCI.—STREPTOCOCCI AND MICROCOCCI,
I, No acidity produced in dextrose or other sugars.

MM, lactis erythrogenes Grotenfelt u.s. A pink, fluorescent coccus. Several
specimens of a micrococcus giving a pink fluorescence have been found. We
have given them the above name, although in our cultures they do not render
milk red. As studied in our laboratary their characters are as follows:

Morphology.—Size, .8u. No chains.

Gelatine colony.—A smooth, flat colony, .5 mm. in diameter in four days,
which later sinks into a shallow pit.

Gelatine stab.—A white needle growth, with a slow liquefaction and a dense
scum, stratiform.

Agar streak.—A luxuriant, white to yellowish growth, witha pink fluorescence.

Fermentation tubes, No acidity and no gas.

Bouillon,—A sediment and a turbidity formed, but no pellicle.

Milk.—Rendered slightly acid and digested into a semi-transparent yellowish
liquid; not curdled.

Potato.—A luxuriant, yellow, moist growth.

Grows at both 20° and 37°. Aerobic.

1
This organism is clearly allied to B. lactis erythrogenes if it is not identical
with it.

M. lactis rubidusn.s. A red coccus, This organism has not been found
since its previous description. It resembles 7. cinnabarius of Fligge, differ-
ing from it in the points shown in brackets. The following is the description
of our organism as given in our previous publication:

Morphology.—Size, 4. No chains.

Gelatine colony.—A rapidly liquefying, red colony. Some of the colonies on
the same plate are not red. [Dull red color.]

Gelatine stab.—Infundibuliform growth, which later liquefies completely,
forming a red liquid. [Liquefaction only partial.]

Agar streak.—A thick, moist, blood-red growth. [Yellowish brick-red.]

Bouillon,—A sediment and turbidity, but no pellicle; the sediment is pinkish.

Mitk.—No change in the action. Old cultures show a digestion with a
pinkish surface.

Potato.—A very luxuriant, blood-red growth [yellow to red] spreading over
the potato.

Grows at both 20° and 37°. Produces no pigment at the latter temperature,
Aerobic,

While this organism is quite similar to Fliigge, his is too insufficiently
described to be sure of the identity.

M. lactis citronus n.s. An orange-colored, liquefying Micrococcus. This
was found in the red slime on Camembert cheese.

Morphology.—Size, .8u-.9#. Gram stain irregular; no chains.

Gelatine colony.—A yellowish, slowly-liquefying colony, with a clear liquid.
Litmus gelatine is reddish-brown in color.

Gelatine stab,—Liquefaction begins in .four days and is never complete.
Stratiform,

118 STORRS AGRICULTURAL EXPERIMENT STATION.

Agar streak.—A spreading, thick, smooth growth of an orange color, some-
what viscous.

Fermentation tubes.—No acidity, no gas, no closed arm growth in any sugar
bouillon.

Bouillon,—A sediment, turbidity and a pellicle.

Milk,—No action on milk.

Potato,—Luxuriant, thick, smooth, orange-brown.

Grows at 37” Aerobic.

S. lactis citreus [.n. >. A lemon-yellow, liquefying Streptococcus.

Morphology.—A streptococcus. Size, .64-14. Gram stain positive.

Gelatine colony.—A small colony, with clear liquid and slow liquefaction.
Not characteristic.

Gelatine Stab.—A slow liquefier with stratiform liquefaction.

Agar streak.—A luxuriant, thin, lemon-yellow growth,

Fermentation tubes.—No acidity, no gas, no growth in closed arm in any
sugar bouillon.

Bouillon.—A turbidity, a sediment, and a pellicle.

Milk.—Slightly alkaline, or no change in reaction, and a slight digestion.

Potato,—Luxuriant, moderately thick, lemon-yellow.

Grows best at 20°; less luxuriantly at 37° Aerobic.

The culture from which this was taken was from the centre of a cheese.

Variety A.—A second culture from the same cheese near the surface, differed
in producing an acidity in dextrose and saccharose, and no pellicle nor diges-
tion of milk.

Variety &.—From Storrs; foundin milk. Did not liquefy the gelatine plate,
but produced a thin, flat colony, 2.5 mm. in diameter.

Fartety C.—From milk in Middletown. Produced a slowly-liquefying,
granular colony, 2-3 mm. in diameter; a wrinkled growth on agar; a pellicle
upon bouillon, and a digestion of milk.

Varie‘y D.—Showed no potato growth, but an acidity in milk, although the
milk digested.

Variety £.—¥rom cheese. Showed no potato growth and a very slow lique-
faction, with no milk digestion. We have no hesitation in regarding them all
as belonging to the same type.

S. lactis Rogertn, 8s. A lemon-yellow, liguefying Streptococcus,

Moi phology.—Size, .7v-I4. Gram stain irregular; chains produced.

Gelatine colony.—A slow liquefaction, with a thin, transparent, granular
colony, at first white. Lzémus gelatine is intensely alkaline, with a spindle-
shaped colony.

Gelatine stab.—A needle growth and a stratiform liquefaction beginning in
one day.

Agar streak,—A luxuriant, thick, lemon-yellow to brown growth.

Fermentation tubes.—No acid, no gas, no growth in closed arm in any sugar
bouillon.

Bouillon.—A sediment and turbidity.

CLASSIFICATION OF DAIRY BACTERIA. 11g

Milk.—Rendered alkaline. Sometimes there is a curdling and sometimes
not, but the milk always digests.

Potato.—Luxuriant, yellow to lemon-yellow, with a discolored potato.

Grows best at 20°; slightly at 37°. Aerobic.

This was very abundant in a Camembert cheese sent us from France. It
differs only slightly from the last described type, being intensely alkaline on
litmus gelatine, and having a very luxuriant potato growth. The two are
closely allied and are perhaps the same.

M. lactis minutissimus, A minute, liguefying Coccus. Has been found but
once, from milk in Middletown.

Morphology.—Size, .2u-.34. Gram stain negative.

Gelatine colony.—Round, thin, smooth, in a clear, liquefying pit. Liquefac-
tion slow.

Gelatine stab.—An infundibuliform liquefaction, with a granular layer on the
surface, or sometimes only a deep, dry pit.

Agar streak.—Scanty, thin, smooth, yellow to lemon yellow.

fermentation tubes.—Lactose is acid; the other sugars are not acid, and no
gas nor closed arm growth is produced.

Bouillon.—A sediment and turbidity, but no pellicle.

Milk,-—No change in the reaction, but the milk is promptly curdled and then
digested, with a prominent odor.

Potato.—A white, dry, wrinkled, luxuriant growth.

Grows best at 20°; slightly at 37°. Aerobic.

M. lactis aureus An. s. A yellowish, liguefying Micrococcus, This organ-
ism is very much like MW. lactis varians, and may be the same. (See p. 121.)
The only essential difference is the lack of acid production, and on this ground
we separate them, although we regard them as allies, or identical. This type is
comparatively rare, while the vardazs type is very abundant. Certain varia-
tions from the described type are found in other cultures, and are indicated in
brackets.

Morphology.—Size, 84-1". Gram stain positive.

Gelatine colony.—A V-shaped colony is formed in a pit surrounded by a halo.
It is irregular, globate or entire; later it liquefies.

Gelatine stab.—A stratiform liquefaction, with a yellow sediment. Liquefac-
tion begins on the second day and is complete in fourteen days.

Agar streak,—A luxuriant, brownish-yellow growth. [White with yellowish
tinge. ]

Fermentation tubes.—No acidity nor gas is produced.

Bouillon.—A pellicle and turbidity, with no sediment. [Sediment and no
pellicle.]

Milk,—Alkaline and curdled; the milk subsequently digested into a clear
liquid, with a half inch of sediment. [No digestion. ]

Potato.—A \uxuriant, brownish-yellow growth.

Grows at both 20° and 37°. Aerobic.

Two types of yellow cocci, unnamed, described by Freudenreich, belong here.

120 STORRS AGRICULTURAL EXPERIMENT STATION.

The white liquefying cocci are very numerous and we have studied many
separate cultures of them. Whether they should all be grouped together is un-
certain. We have endeavored to arrange them in sub-groups, as shown below,
but are convinced that these represent only physiological varieties of the same
type. Weare also of the opinion that this group is simply a liquefying form
of the white coccus, Gal. versicolor. (See p. 110.) :

M. lactis albus n. ».

Morphology.—Size, .7u-1". Gram stain is positive; no chains.

Gelatine colony.—Round, moderately luxuriant, thick, smooth colonies, which
may liquefy or in some cultures do not liquefy. In Utmus gelatine the colony is
dense and alkaline.

Gelatine stab.—A very slow liquefier, that commonly forms a dry pit with a
white growth on its sides.

Agar streak.—Moderately luxuriant, opaque, whitish.

Fermentation tubes.—No acidity, gas, or closed arm growth in any sugar
bouillon. :

Bouillon.—A very slight growth, showing a slight turbidity and a sediment
but no pellicle.

Milk.—Rendered alkaline and digested without curdling. Later the milk
becomes pinkish and slimy.

Potato,—Luxuriant, thick white.

Grows at 20° and 37°. Aerobic.

Variety A.—Shows a viscosity on agar and sliminess in milk.
Variety B.—Shows no viscosity; a wrinkled growth on agar; a pellicle on
bouillon and a discolored potato. Milk is curdled at 37°.

Variety C.—Has a moruloid colony that slowly liquefies, produces a pellicle
on bouillon, and no milk digestion.

Variety D.—Liquefies more rapidly and curdles milk.
Variety E.—A very large coccus, 24 in diameter.

Variety F.—A very slow liquefier, curdling and digesting milk; the milk
after some weeks turning to a dark mahogany color. Potato growth is dry and
velvety. <

To this same group belong doubtless the small coccus 4. of Freudenreich.

II. Acid in dextrose or other sugars.

M., lactis fluorescens n.s. Only one fluorescens coccus has been found, in
stable dust.

Morphology.—Size, .5u-.6u. Gram stain negative,

Gelatine colony.—A round, moderately thick, smooth, entire colony, with a
greenish liquefaction. Litmus gelatine shows liquefying pit that is not acid,

‘Gelatine stab.—A stratiform liquefaction.

Agar streak.—A luxuriant, narrow, rather thick, smooth, white growth, with
a green fluorescence.

Fermentation tubes.—Dextrose is rendered acid, the other sugars alkaline: no
gas is produced. Growth appears in closed arm.

Bouillon.—A sediment, turbidity, and pellicle.

CLASSIFICATION OF DAIRY BACTERIA. 121

Milk.—Rendered slightly acid, curdled, and later completely digested into a
greenish-yellow liquid. ‘

Potato.—Scanty, thin, smooth, white.

Grows at both 20° and 37°. Facultative anaerobic.

M. lactis varians n. s. Yellow, liquefying, acid-producing Cocci. This is
perhaps the most common and widely diffused type of coccus found in milk.
We have found it in milk from many localities, and it is very abundant. It is
frequently present in milk directly from the udder. It shows a wide range of
variations affecting nearly every character. We have studied some scores of in-
dependent cultures from different sources, and find that the different types
grade into each other by such slight differences that we have no hesitation in
putting them all together as one type. This organism liquefies gelatine, but in
some cases so slowly as to form only a dry pit. From this it is only a step to
a non-liquefying form. We, therefore, are inclined to believe that this type is
identical with, or at least closely allied to, MW. lactis aureus (p. 112). The
characters as we have studied them agree essentially with those of Staph. pyogenes
aureus. Cultures of the latter organism, sent us by Retger, have been com-
pared side by side with our cultures and no essential differences are seen. We
are inclined to think, therefore, that our type is the common Staph. pyogenes
aureus. In the description that follows the limits of variations will be given
under each head and no attempt will be made to distinguish distinct varieties.

Morphology.—Size, .4u-1.44. Gram stain positive.

Gelatine colony.—Deep colonies are opaque; surface colonies form white or
yellowish beads or a slow liquefying colony with a clear liquid and a granular,
mottled, irregular growth, distributed in the liquid. It is usually slightly acid
in litmus gelatine, sometimes decidedly so, forming a bright red liquid colony.

Gelatine stab.—Liquefaction is usually slow but sometimes rapid. It com-
monly begins in from two to four days, but is never complete. In some cul-
tures liquefaction is so slow that only a dry pit is formed, with a broken,
yellow growth on its side. It is usually napiform.

Agar streak.—A luxuriant growth, frequently tending to be rough, though
never wrinkled. It does not spread profusely. Its color is typically pale
orange-yellow, but varies from this to nearly orange and to practically white.
In the latter case this is quite indistinguishable from JZ. lactis albidus (p. 123).
The agar growth may be dry or moist.

Fermentation tubes.—All three sugars are rendered acid and growth appears
in the closed arm. No gas is ever formed. Some cultures do not grow in the
closed arm and some have failed to render lactose and saccharose acid.

Bouillon.—A flocculent sediment is produced and a slight turbidity. Two
cultures showed a granular pellicle.

Milk,—Is always rendered acid and commonly curdled and digested with a
yellow sediment. In the non-liquefying cultures the curdling may not appear
nor any noticeable digestion.

Potato.—Is usually luxuriant, though sometimes scanty, of a pale orange-
yellow color, showing the same variations as mentioned in agar streak. It is
frequently dry.

Grows better at 37° than at 20°. Facultative anerobic.

122 STORRS AGRICULTURAL EXPERIMENT STATION.

These variations clearly run into AZ. /actis aureus on one hand and &M. éactis
albidus on the other, so that a sharp separation is impossible. This corre-
sponds to M/ic. Z. of Barthel and is probably everywhere distributed.

This organism is‘not only common in this country but it appears to be com-
mon in Europe as well, several described forms doubtless belonging to the
type, for example, Stad/-luftbacterium JZ, of Koning and others. Koning’s
Stall-luftbacterium IZ., which he regards as different, shows such slight varia-
tions from this that it also comes within the limits that may properly be regarded
as covered by our MW. lactis varians.

M. lactis varians A.—Under this head we recognize two cultures derived
from milk fresh from the udder, agreeing with the type in all points except the
following:

Acid is produced in dextrose only.

Milk is not rendered acid, although it may be curdled and digested.

We do not think this is sufficiently different to constitute a new type, but
tabulate it here separately.

M., lactis giganteusn.s, An extremely large, iquefying Coccus.

Morphology.—Size, 1.44-3.54. A micrococcus which accepts the Gram stain.
Its peculiarity is the very large size which the coccus sometimes reaches—3.5u
in diameter.

Gelatine colony.—A clear, liquefying pit, which is slightly cloudy and white.
Litmus gelatine is not acid.

Gelatine stab.—Begins to liquefy in one day, infundibuliform.

Agar streak,—A moderately luxuriant, smooth growth, of an orange color.

Fermentation tubes.—All sugar bouillons are made acid, but there is no gas.
nor closed arm growth.

Bouillon.—A sediment, but no turbidity nor pellicle.

Milk.—Rendered acid and digested into a yellow liquid at both 20° and 37°.

Potato.—A scanty, beady, brownish growth.

Grows at both 20° and 37°. Aerobic.

This agrees with the last in its physiological properties, but the extraordinary
size of the cells is so peculiar that we have given it independent rank.

M, lactis rugosus n. >. A salmon-yellow Coccus. Perhaps M. acidi lactici of
Kriiger.

Morphology.—Size, 1u¢-1.2u. Gram stain irregular; micrococcus.

Gelatine colony.— A liquefying pit, with a clear centre and a ring of granular
matter; white.

Gelatine stab.—A slow liquefier, crateriform or stratiform.

Agar streak.—A salmon-yellow growth, luxuriant and viscous, wrinkled,
with a dull surface and a salmon-yellow color.

Bouillon.—A sediment, turbidity, and a ring-like pellicle.

Milk.—Rendered acid and curdled, with an orange color and a sour odor;
no digestion.

Grows well at 20° and 37°. Aerobic.

CLASSIFICATION OF DAIRY BACTERIA. 123

We have found this only once. Kriiger apparently found the same in milk
and butter. His organism differed from ours in digesting milk. The salmon
color gives it independent rank.

The white, liquefying, acid-producing cocci form another series of extremely
common Bacteria, with a long list of variations. As already mentioned, they
pass, by imperceptible grades, into the yellow acid cocci, and should, perhaps,
be united with them. These white cocci appear identical in cultural characters
with Staph. pyogenes albus, a culture of which, when compared side by side,
showed no essential differences. Staph. mastitis albus of Guillebau seems also
to be essentially the same, as well as four different varieties of white cocci de-
scribed by Freudenreich. (A/zlch2tg. 1905, pp. 628 and 643.) These white
cocci are extremely common in milk, in the udder, in the dust of the stable,
and have been found in many samples of milk here and elsewhere. Among the
many variations some are quite striking, and we have, therefore, endeavored to
separate the various strains studied in groups, as follows:

M, lactis albidus n. s.

Morphology.—Micrococci. Size, .6u-1.24. Stains by Gram method.

Gelatine colony.— An opaque colony, usually white, and soon liquefying. It
is not characteristic. On Utmus gelatine it is sometimes acid and sometimes
not acid.

Gelatine stab,—Liquefies in from one to three days, infundibuliform. Some-
times a dry pit is formed, which may liquefy after several days.

Agar streak.—A moderately luxuriant, smooth, white growth; not very thick.

Fermentation tubes,—All sugars are rendered acid but no gas is formed, and
there is usually no closed arm growth. (One culture did not produce acid or
grow in bouillon.)

Bouillon.—A sediment and turbidity, but no pellicle.

Milk,—Usually rendered acid, and may or may not curdle. Curdling is
more common, however, than not curdling. The milk is digested, except in
those cultures that do not liquefy gelatine. One culture, however, liquefies
gelatine, but does not curdle milk.

Potato.—A moderate growth, white to yellow; not characteristic.

Grows at 20° and 37°. Facultative anaerobic.

To this type belongs the white cocci of Guillebau and Freudenreich.

Variety A.—This differs from type Variety A chiefly in producing a snow-
white growth on agar. Two cultures were studied, one of which produced a
snow-white colony in gelatine, and the other lacked snow-white color. One of
the two was snow-white on potato, while the other produced a thin, scanty
growth. In all other respects they resemble the type.

Variety B.—This is separated from the others by its more anaerobic charac-
ter, as shown by growth in the closed arms of the fermentation tubes. The
colonies on litmus gelatine are acid; the agar growth is scanty, as is also the
growth on potato. This was foundon Cheddar cheese and Camembert cheese.

Variety C.—Distinguished from the others chiefly by not making milk acid
and by not curdling. One culture of this variety renders the milk alkaline,

124 STORRS AGRICULTURAL EXPERIMENT STATION.

curdles and digests it. The acidity in sugar bouillon is less pronounced than
in the other varieties, and in lactose there is usually no acidity. The litmus
gelatine colonies are usually acid, though not always. In other respects they
agree with the type.

M. Freudinreichii of Guillebeau appears to belong here, differing from those
mentioned above in rendering milk sdmy.

THE GENUS SARCINA.

We have not found the genus sarcina very commonly represented in milk
products. The cultures which we have found may all be reduced to four types;
for, while the different specimens show some variations, they are not very great,
and not so great but what they may be properly included under one of the four
types.

Sar, lactis albusn.s. A white or yellow non-liquefying Sarcina.

Morphology.—Size; .74. The Gram stain is positive and there is no motility.

Gelatine colony.— Round, convex, smooth, homogeneous, entire, yellowish or
white. —

Gelatine stab.—A needle growth and a convex surface growth,

Agar streak.— Beaded, raised, smooth, translucent, cream-white, moist, not
luxuriant,

Fermentation tubes.—Acidity and closed arm growth in all sugar bouillons,
but no gas. Sometimes lactose shows no closed arm growth.

Boutllon,—A sediment and a slight turbidity, but no pellicle.

Milk.—Becomes sufficiently acid to curdle on boiling. No other change.

Potato.—A very slight cream-white growth.

Grows at 20° but scarcely at all at 37° Facultative anaerobic.

. Sar, lactis lutean.s. A yellow, liguefying Sarcina, Resembles Sar. lutea
of Fligge.

Morphology.—Size, .74-1p. Gram stain positive. Not motile.

Gelatine colony.—A slow liquefying pit, with a nucleus surrounded by
granular area; yellowish. Litmus gelatine not acid.

Gelatine stab.—Begins to liquefy in about three weeks, crateriform, and never
complete.

Agar streak.—Filiform, raised, smooth, opaque, lemon-yellow, luxuriant.

fermentation tubes.—No acidity, no gas, no closed arm growth in any sugar
bouillon.

Boutllon.—A sediment, but no turbidity nor pellicle.

Milk.—Becomes alkaline and slowly digests, with a yellow color, but does
not curdle.

Potato.—Beaded, raised, opaque, lemon-yellow, luxuriant.

Grows at both 20° and 37° _— Aerobic.

Among the variations found in our several cultures the following may be
mentioned: Size, from .74-24; color, lemon-yellow to whitish; the reaction on
milk is frequently not changed; the growth on potato ranges from luxuriant to
no growth.

CLASSIFICATION OF DAIRY BACTERIA. Las

Sar, lactis aurantiaca n. ». An orange, ligquefying Sarcina, This may be
the same as the last, but its color is quite different and it shows some other
differences. Probably Sar. aurantiaca of Fligge.

Morphology.—Size, 1m. Gram stain positive, non-motile.

Gelatine colony.—A liquefying pit, forms with an orange pigment. Litmus
gelatine is not acid.

Gelatine stab.—A slow liquefaction, stratiform.

Agar streak,—Filiform, raised, smooth, moist, orange, luxuriant, slightly
viscous.

Fermentation tubes.—No acidity, no gas, no closed arm growth in any sugar
bouillon.

Bouillon.—A sediment, a membranous pellicle and a slight flocculent sedi-
ment.

Milk, No change in reaction, or a slight alkalinity; the milk is curdled and
digested. Digestion is nearly complete, with an orange sediment.

Potato,—Spreading, capitate, contoured, orange color, luxuriant; potato dis-
colored.

Grows at both 20° and 37° —_ Aerobic.

Sar. lactis acidin. ». An acid, yellow Sarcina.

Morphology.—Size, .8u-14. Gram stain positive; not motile.

Gelatine colony.—Round, raised, smooth, homogeneous, opaque, brownish,
slowly liquefying.

Gelatine stab.—Liquefaction very slow and sometimes only a dry pit is
formed, with a yellow bacterial growth.

Agar streak.—Filiform, raised, smooth, cream-white, moist, not very luxuri-
ant. Color is sometimes yellow.

Fermentation tubes.—Acidity produced in all three sugar bouillons (it may be
lacking in saccharose), but there is no gas nor closed arm growth.

Bouillon.—A sediment and a slight turbidity, but no pellicle. (Turbidity
sometimes absent.) :

Milk,—Becomes acid, but does not curdle or digest.

Potato.—Toes not grow well, but there is a slight cream-colored growth.

Grows at both 20° and 37°, though not very abundantly at 37° ‘Aerobic.

THE GENUS BACTERIUM, NON-LIQUEFYING,
I. No acid in dextrose or other sugars.

Bact. lactis salmonis n.s. A salmon-colored Bacterium.

Morphology.—Size, .64x 1¢-1.84, forming chains. Gram stain is positive and
there are no spores nor capsules.

Gelatine colony.—Round, umbonate, contoured, lobed, white. Litmus gelatine
colonies are strongly alkaline and transparent.

Gelatine stab,—Needle growth and a raised surface growth.

Agar streak,—Filiform, thin, smooth, white to yellow, and later a salmon or
pink color.

Fermentation tubes.—No acidity, gas, nor closed arm growth in any sugar
bouillon.

126 STORRS AGRICULTURAL EXPERIMENT STATION.

Bouillon.—A flocculent sediment, a membranous pellicle and a slight tur-
bidity.

Milk.—Becomes alkaline, but shows no other change.

Potato.—Luxuriant, thick, contoured, flesh color or pink.

Grows well at 20°; slightly at 37°. Aerobic.

This was found by Harding in a specimen of green butter.

Bact. lactis aureum I, An orange, non-acid Bacterium, Perhaps B. lac-
tericus of Adametz.

Alorphology.—Rods which do not form chains. Size, .74¥-.g@x IM¢-34. No
spores are produced, a capsule is evident and Gram stain is positive.

Gelatine colony.—Round, flat, contoured, lobed, orange to yellow color.
Litmus gelatine colonies are of a red-brown color. ‘

Gelatine stab.—A needle growth and a thin, reddish surface growth.

Agar streak,—\uxuriant, deep orange-brown color, tough and tenacious,
sometimes dull, and aggregated in colonies.

Fermentation tubes.—No acidity, gas, nor closed arm growth in any sugar
bouillon.

Bouillon.—A turbidity, sediment, and a pellicle.

Milk.—No action except a slight orange color at the surface.

Potato.—Growth scanty or absent; when present, of an orange color.

Grows better at 20” degrees than at 37°. Aerobic.

Found in Middletown and in New York city.

Bact. lactis citreum If. n.s. A yellow, non-acid Bacterium.

Morphology. A rod occasionally forming chains. Size, BUH." X v7UHT Ape
No spores nor capsules, and Gram stain negative.

Gelatine colony.—A round, opaque bead, 1.5 mm. in diameter; usually white
and later turning yellow.

Gelatine stab.—A scanty needle growth and an irregular, dry, white surface.

Agar streak.—Luxuriant, at first white, but soon lemon-yellow.

Fermentation tubes.—No acidity, gas, nor closed arm growth in any sugar
bouillon.

Bouillon.—Turbidity, sediment, and a flaky scum.

Milk.—No action.

Potato.—Luxuriant, yellow, thick, sometimes wrinkled.

Grows both at 20° and 37°. Aerobic.

Variety A.—This has a small lemon-yellow colony on gelatine which is
lemon-yellow even when viewed under the microscope. It has also a lemon-
yellow surface growth on the gelatine stab and a lemon-yellow growth on
potato. .

Variety B.—Produces a yellow pigment, but not lemon-yellow, both in
gelatine colony and on potato. It is slightly viscous on agar, and has no pel-
licle on bouillon. Potato is slightly discolored, and one culture failed to grow
on potato. All from milk.

CLASSIFICATION OF DAIRY BACTERIA. 127

The white non-acid bacteria are very numerous and show many slight varia-
tions. We have tried to separate them into groups, but they are all more or

less connected by intermediate forms and our grouping is not very satisfactory.
a

Bact, lactis myceloidium. A mycoloid, non spore-bearing Bacterium, We
have studied two cultures of this type, one from this locality and one sent us
by Weigmann, from Kiel, as Bact. mycoides. The Kiel culture is not mycoides,
for it fails to produce spores and does not liquefy gelatine. This raises the
question whether it may not be a cultural variety of 2. mycoides, having lost
these two properties. As tested in our laboratory, it had the following charac-
ters:

Morphology.—Long filaments, the individual elements of which are .7u x 2u—
3.5@. The Gram stain is irregular and there are no spores,

Gelatine colony.—A myceloid colony, 2 cm. in diameter, spreading rapidly;
largely under the surface.

Gelatine stab,—A needle growth and a surface growth; a layer of threads is
seen extending horizontally, a short distance below the surface, to the sides of
the tube.

Agar streak.—A luxuriant, moderately thick, slightly yellowish growth.

Fermentation tubes.—No acidity, gas, nor closed arm growth in any sugar
bouillon.

Bouillon.—A sediment and turbidity, but no pellicle.

Milk.—Becomes slightly acid, but does not curdle, even when heated, and
shows no other change.

Potato.—A scanty growth, thin, white.

Grows better at 20° than at 37° Aerobic.

We have found a local variety of the above, agreeing with it in most respects.
It is shorter, .9 #, does not show the peculiar horizontal growth in gelatine stab,
and produces no acidity in milk.

Bact. lactis arborescens J, n.s. An arborescent, non-acid Bacterium.

Morphology.—Size, .o#X 1.2u-1.4#. It has no spores nor capsules, forms no
filaments, and Gram stain is negative.

Gelatine colony.—Round, raised, smooth, entire, white colony. On litmus
gelatine brownish and not acid.

Gelatine stab.—An arborescent needle growth and a surface growth.

Agar streak.—Scanty, thin, white to cream color, slightly viscous.

Fermentation tubes.—No acidity, gas, nor closed arm growth.

Bouillon.—A sediment, ring-formed pellicle, and a slight turbidity.

Mitk.—Is rendered alkaline and slimy. After some days it becomes slightly
transparent, indicating a slight digestion.

Potato.—Scanty, raised, grayish-brown color; potato discolored,

Grows at both 20° and 37°. Aerobic.

This organism was sent us from Michigan by Marshall, but we have not @ur-
selves found it. Its slight digestion of milk and its slight pit in gelatine stab
suggest an intermediate step toward a liquefying form. If regarded as a slow
liquefier it is not unlike Bact. arborescens Frankland, found in water.

1

128 STORRS AGRICULTURAL EXPERIMENT STATION.

Bact. lactis viscosum Adametz. Under this name are included several slimy
milk bacteria described by different observers. The first was described by
Adametz, others by Ward, Harrison, Freudenreich, and Marshall. We have
had an opportunity af studying all of these except that of Adametz, from
original cultures from the authors. We have also received a similar culture
from New York city milk. These have all been carefully studied by Harrison
who regards them as a single type and calls them group 1. We are in agree-
ment with him in recognizing this as a logical group. Its general characters
are as follows:

Morphology.—Size, .5u-1.2¢x.5u-2.5. Frequently narrower at the ends.
Forms filaments, 154 in length. The slime seems to be produced from a
capsule, but this is not always seen. Gram stain negative.

Gelatine colony.—F lat colonies, with irregular edges, or lobate. 3 mm.—6mm.
in diameter. Later viscous.

Gelatine stab,—A good needle growth, which may be separated into granules,
Sometimes arborescent, but not always so. A thin, shiny, gray surface growth,
lobate.

Agar streak.—Usually luxuriant, viscous, white, not very thick,

Fermentation tubes.—No acidity, gas, nor closed arm growth in any sugar
bouillon.

Bouttlon.—Turbidity, pellicle, and sediment. (The organisms of Ward,
Harrison, and Freudenrich show no pellicle.)

Milk.—Becomes alkaline, does not curdle, but is very slimy.

Potato,—A thick, uneven, dirty gray, becoming brown or yellow; slimy.

Grows at both 20° and 37°. Aerobic.

We retain the name of Adametz, changing the word Bacillus into Bacterium.
I see no good reason for separating the above mentioned organisms even as
varieties. Many cases of slimy milk infections in dairies are produced by this
type of bacterium, which is probably the most common cause of such troubles.

Bact. lactis acidi, Var. E. This organism is very similar to Bact. acidi
lactici (see page 134). In all of its characters, except one, it agrees with that
type. It shows the same unwillingness to grow in common culture media in
the laboratory, and little or no surface growth, but it produces no acidity in
sugar bouillons and no acidity in milk. Inasmuch as Bact. acidi lacticd shows
great variations in the power of producing acids, we regard this as an extreme
variety of that type. No further description here is necessary. .

Bact. lactis Connii Chester. A white non-acid Bacterium. Large numbers of
common white bacteria have been found without striking characters. They
are common in all milk. We have found them more or less constantly here
and in New York milk. They are also common in cheese, and constitute the
chief bacterium in the red slime of Camembert cheese. In our former report
we recognized two or three different types among them, but at present we
believe they should all be grouped together under one head. We have retained
the name given by Chester, It is parallel with Galactococcus versicolor among
the cocci; it may be identical with the latter.

CLASSIFICATION OF DAIRY BACTERIA. 129

Morphology.—A bacterium (.54-.7“x 1.4“), forming short chains. It has no
spores nor capsules and the Gram stain is negative.

Gelatine colony.—Round, raised, smooth, entire, white or cream color, Li/-
mus gelatine shows a non-acid, white, not characteristic surface colony.

Gelatine stab.— A good needle growth and a white surface.
Agar streak,—Luxuriant, filiform, raised, smooth, white, opaque.

Fermentation tubes.—No acidity, gas, nor closed arm growth in any sugar
bouillon.

Bouitlion.—A sediment, ring-formed pellicle, and slight turbidity.

Mdilk.—Rendered slightly alkaline, is not curdled, and shows no digestion.
In one case milk became pasty in about three weeks.

Potato.—Luxuriant, convex, smooth, white, potato discolored.
Grows both at 20° and 37°. Aerobic.

Among the numerous cultures which we have studied we can recognize two
sub-varieties.

Variety A.—A less vigorous organism than the one described. Size, 1.4ux
1.24, forming rather long filaments. Bouillon sometimes shows no pellicle
and milk is not rendered alkaline. Potato growth is rather scanty. The potato
shows no discoloration, but the growth itself is slightly yellowish.

Variety B.—Stains by the Gram method; agar streak is dry and wrinkled;
dextrose slightly acid; no discoloration of potato.

This seems to be identical with B. /actarius of Adametz aod. a Bacterium
described by Burri and Dugelli, mentioned as having a ‘‘dog odor” appears
also to be the same.

Il. Acid in Dextrose or other Sugars.

Bact. rudensis Connelli. A red, acid Bacterium, This organism was isolated
by Harding from cheese vats, and regarded by him as the above species. Four
cultures were sent to us by him after he had kept them in stock fora year. Of
these four cultures two came originally from Canada; two of them had entirely
lost their power of producing red pigments, and two still gave a slight red color
to milk. All were said to produce red pigment originally. The cultures which
I received had also totally lost their flagella and motility, and in this condition
were not distinguishable from 8. /actis acidi, except by showing a better
growth on potato. These cultures, when studied by us, had the following
characters.

Morphology.—A short rod, 1#x 1.84; forming no chains, showing no spores,
and staining by the Gram method (originally motile).

Gelatine colony.—A small colony, mostly under the surface, quite dense. On
litmus gelatine it is intensely acid.

Gelatine stab,—A needle growth, but no surface growth (originally a thin
surface).

Agar streak.—No visible growth, or sometimes an extremely thin tiauepmnent
growth.

130 STORRS AGRICULTURAL EXPERIMENT STATION.

Fermentation tubes.—Acidity and closed arm growth in all sugar bouillons,
but no gas.

Bouillon.—A sediment and slight turbidity, but no pellicle.

Milk,—Becomes acid and curdles promptly at both 20° and 37°. A rusty
precipitate appears in some cultures.

Potato.—Scanty, reddish-brown color (originally showing yellowish colonies
that become red).

Grows at both 20° and 37°. Facultative anaerobic.

If this colorless variety should be met incidentally in milk, it would never be
thought to be Bact. rudensis, and would doubtless be placed with Bact. /actis
acidi group, and perhaps be regarded as identical with that organism. ‘This
raises the question naturally whether other members of that group may not be
albino types of pigment-producing bacteria. So far as we know we have never
found this species, although we have no proof that some of our white types are
not such albino varieties.

Bact, lactis catenensisn.s. Yellow spore-bearing Bacterium.

Morphology.—Size, .7u x 1.2u; producing chains. Gram stain is negative,
and small spores are produced. In old cultures the rods may be much smaller,
5 be

Gelatine colony.—Round, raised, smooth, homogeneous, transparent, and of a
yellow to white color. On ¢mzs gelatine the color is yellow to gray, not acid.

Gelatine stab,—A good needle growth, and a flat orange-yellow surface.
Agar streak. —Orange to yeliow, thin, and sometimes wrinkled.

Fermentation tubes.—Dextrose and lactose acid, saccharose not acid. No gas
or closed arm growth.

Bouillon.—A sediment, turbidity, and a pellicle.
Milk.—No effect, as a rule, but one culture curdled milk, acid at 37°.
Potato,—Luxuriant, wrinkled, orange-yellow.

Grows at 20° and 37°, but better at 20°. Aerobic.

Three cultures of this general type have been studied from Middletown,
Cromwell and New York. The first was orange, the second lemon-yellow, and
the third brown-orange on agar, and white on potato and gelatine. The last
did not show a wrinkled growth on agar, although it did on potato.

Bact, lactis aureum I], n.s. An orange, acid-forming Bacterium,

Morphology.—Size, .8u-1.2ux 1,2u-1.84. No chains, no spores. Gram stain
negative.

Gelatine colony.—Round, convex, smooth, entire, orange-yellow. On tmus
gelatine it is thin and not acid.

Gelatine stab.—A needle growth and a thin surface growth.

Agar streak.—Moderately luxuriant, filiform, thin, smooth, orange color,
moist.

CLASSIFICATION OF DAIRY BACTERIA. I31

Fermentation tubes.—Dextrose acid, lactose and saccharose slightly acid; no
gas nor closed arm growth.

Bouillon.—No visible growth. i

Milk.—Very slightly alkaline, no curdling and no digestion.

Potato,—Spreading, thin, smooth, yellow.

Grows better at 20° than at 37°. Aerobic.

This organism was sent me by Gorini from Italy. Another, which seemed
to be identical, was found in milk here, the only points of difference being a
luxuriant potato growth and a slight sediment in bouillon. A very similar cul-
ture was sent by Harding, differing only in having a somewhat dry, wrinkled
growth on agar. We regard the three as identical.

Another culture is very closely related to this, but differing in enough par-
ticulars to lead us to regard it as a separate variety.

Variety A.—This organism was found here on Camembert cheese and
another culture was sent us by Gorini. It differs from the above described
type only in the following points: Size, .qu-1.5#x.5. Gram stain positive,
grows well in bouillon, with a sediment. Milk distinctly alkaline. Color of a
lemon instead of an orange yellow.

Variety B.—This organism, sent by Harding, differs from the type in pro-
ducing a rather more orange color, a turbidity and sediment in bouillon, and in
making milk distinctly acid, but not curdling it.

Bact, lactis synxanthum.—A culture of this organism, several years old, was
sent me by Harrison. It had completely lost its power of producing yellow
pigment. Below are given its characters as made out by us, and in brackets
the characters as originally described where they differ from those observed.

Morphology.—Size, .8u-.guxi.24-2u. No spores, no chains. A capsule is
evident. Gram stain is negative. [Gram stain positive, motile. ]

” Gelatine colony.—Round, capitate, smooth, homogeneous, entire, opalescent,
gray, moist. [Luxuriant, gray, yellow.] On ditmus gelatine transparent, white
colonies.

Gelatine stab.—A needle growth and a raised surface growth.

Agar streak.—Luxuriant, filiform, raised, smooth, opaque, porcelain white.
Slightly viscous. [A yellow pigment, soluble in water. ]

Fermentation tubes.—All three bouillons are acid, but there is no gas nor
closed arm growth.

Bouillon.—A sediment, slight turbidity, and a ring-formed pellicle.

Milk.—Becomes acid but does not curdle nor digest. [Alkaline, and digests
to a bright yellow color.]

Potato.—Filiform, raised, contoured, sebaceous, gray, luxuriant; potato
discolored.

Grows at both 20° and 37°. Aerobic.

These organisms show how decidedly characters which are relied upon to dis-
tinguish types may disappear after long cultivation, and naturally throws doubt
upon all classifications based upon physiological properties. 1f an organism
originally isolated because of its power of producing red pigment lose this power
absolutely, and if one which produces a brilliant yellow color loses this property,
we naturally ask whether any physiological properties are constant.

132 STORRS AGRICULTURAL EXPERIMENT STATION.

Bact, seifige Milch Weig. This organism from soapy milk we have had an
opportunity of studying froma culture sent by Weigmann. The culture had
been under observation in his laboratory for some years before it was sent and
did not agree in all its characters with his original description. The descrip-
tion below is of the culture which we have studied and where it differs from
the original description, the latter is indicated in brackets.

Morphology.—Size, 14x .5u. No spores, no capsules. Gram stain negative.

Gelatine colony,—Round, capitate, smooth, homogeneous, entire, translucent,
gray-white. On Utmus gelatine it grows chiefly below the surface and is
strongly acid. After some days the acid reaction changes to alkaline.

Gelatine stab.—A needle growth and a raised surface growth, which later be-
comes a dry pit [slowly liquefying].

Agar streak.—Filiform, thin, smooth, opalescent, gray, moist, not luxuriant,
[Of a yellow color.]

Fermentation tubes.—All three sugar bouillons become acid, but there is no
gas and noclosed arm growth.

Bouillon.—A sediment and turbidity, but no pellicle.

Milk.—Becomes amphoteric and a slight yellow scum appears around the
rim. [Originally this produced a soapy taste in milk, but did not do so when
studied by us. ]

Potato,— Filiform, thin, smooth, brownish, moist, luxuriant [yellow].

Grows at both 20° and 37° [best at 10° according to Weigmann]. Aerobic.

Bact. lactis signiin. ». This bacterium has the unusual character 9 com-
pletely digesting milk into a transparent liquid without previous curdling, but
not liquefying gelatine. It is the only species we have found showing this
character. It was found upon an Isigny cheese and constituted about 44% of
the bacteria on the cheese.

Morphology.—A small rod, not forming chains. Size, .5u-.34. It forms no
spores nor capsules, and the Gram stain is negative.

Gelatine colony.—Round, raised, smooth, homogeneous, entire, opaque, yel-
lowish, moist.

Gelatine stab.—A filiform needle growth, and a flat surface.

Agar streak,—Luxuriant, filiform, raised, smooth, translucent, yellowish,
moist.

fermentation tubes.—After three days an acidity in all three bouillons,
but no gas and no closed arm growth.

Bouillon.—A sediment, slight turbidity, but no pellicle.

Milk.—Becomes slightly acid and digests. After two months the digestion
is complete.

Potato.—Spreading, flat, smooth, cream-white, luxuriant.

Grows at both 20° and 37°. Aerobic.

Bact. lactis non-acidi n. >. This organism belongs to the lactis series (see
below), but the departure from the central type becomes so great as to properly
demand a separate name. The surface growth is very abundant and the acid
production very feeble. The dextrose alone is rendered acid. Milk is never
acidified nor curdled. The complete description is as follows:

CLASSIFICATION OF DAIRY BACTERIA. 133

Morphology.—Size, 1.84 x 5u4-1.24. Sometimes long chains are formed.
There are no spores and the Gram stain is irregular, commonly negative, but in
two cultures was positive.

Gelatine colony.—Small, round, moderately thick, entire or irregular margin,
white. Litmus gelatine colony is not acid and sometimes distinctly alkaline.
Sometimes green. ‘The colonies of different cultures show considerable varia-
tion which may indicate different varieties. Our data, however, at present do
not warrant us in separating them.

Gelatine stab.— A needle growth and a spreading, moderately abundant sur-
face.

Agar streak.—Moderate to luxuriant, linear, smooth, white, moist.

Fermentation tubes.—Dextrose acid, and sometimes lactose and saccharose.

Bouillon.—A sediment and turbidity, and frequently, though not always, a
pellicle.

Milk. — Alkaline, or no change in reaction, no digestion nor other change.

Potato,—Scanty to luxuriant, white, spreading. One culture was wrinkled.

Grows at both 20° and 37°, though better at 20°. Aerobic.

The variations given above are large and we feel that this group should be
broken up. But the different variations mentioned cross each other so much
that we have as yet been unable to divide them into any definite varieties. As
more cultures accumulate and give us more information we believe it will be
possible to recognize some sharply distinct types, but at present we leave them
together. The variations which we have found fail to group themselves, and if
we should try to make varieties we should be obliged to recognize nearly as
many as we have isolated individual cultures.

This type of lactic bacteria is very common. It appears constantly in milk in
this vicinity as well as in New York state, and in cheese. It does not seem to
be, however, so vigorous as the typical lactic organism, and is apparently not
the cause of ordinary sour milk.

_ Bact. lactis ubiquitum. This organism we have not especially studied since
its description in 1899. We reinsert here the description then given:
Morphology.—Size, 1.2¢-1.4u x .8u. Long chains are formed; spores are
developed and also a capsule.
Gelatine colony.—Round, capitate, smooth, entire, white, the outer edge
thinner and lighter.
Gelatine stab.—A needle growth and a rather thick irregular surface growth.
Agar streak.—A luxuriant, white surface growth, developing irregular frost-
like or feather outgrowths. Moist, smooth.
_ Fermentation tubes.—Not determined, but probably acidity is produced, at
least in dextrose and lactose, and there is no gas.
Bouillon.—A sediment and turbidity, but no pellicle.
Milk.—Rendered acid and is curdled after several days. No digestion.
Potato.—A luxuriant growth, transparent, spreading, white, glistening.
Grows well at both 20° and 37°. Aerobic.

134 STORRS AGRICULTURAL EXPERIMENT STATION.

WHITE, ACID-PRODUCING BACTERIA.

These are immensely numerous. They are the common cause of the souring
of milk, and since this phenomenon is practically universal, it follows that these
bacteria are equally widely distributed. This group contains the dairy bacteria
par excellence. By this is not meant that these are most abundant around the
barn or in fresh milk. As pointed out elsewhere, they are not common in the
udder of cows, and are usually present only in small numbers in freshly drawn
milk. But they are so much better adapted to life in milk that they soon
become more abundant than all other bacteria put together. Thus in older
samples of milk they are by far the most common.

We have studied hundreds of cultures belonging to this general type, obtained
from all over this country, as well as from several localities in Europe. Among
this large series of samples we have found endless variations. There is hardly
a characteristic which does not show wide variations in the numerous cultures
studied. They include such widely different types that it is hardly proper to
put them into one group, but to arrange any satisfactory subordinate grouping
is almost an impossibility. This might be done by selecting almost any of the
characteristics of the type and arranging the different cultures according to its
variations. The most satisfactory arrangement as appeared to us is to divide
them according to their power of producing a surface growth on various media.
This is not a sharp character, but it presents at least two extremes which may
be clearly separated.

Heinemann (Cent. f. Bac. u., Par. II., XVI., p. 538, 1896) doubts whether
the common lactic organisms should be called by the name Bacterium, believ-
ing that they are really all Streptococci. We are inclined to doubt the correct-
ness of this view, and prefer to retain the long accepted nomenclature.

Bact. lactis acidi Leichmann. This is the common cause of sour milk.
Among the many scores of organisms of this type which we have studied
we have tried to recognize some groups worthy to be called varieties. Whether
this is possible is uncertain, since all are connected by slight intermediate
gradations. We recognize, however, the following:

Bact. lactis acidi, type.

Morphology.—A bacterium. Size, .7u-1.24x .5u-.84. Sometimes so short
as to be described as a streptococcus, and some cultures are very clearly cocci.
(See p. 114.) There is no motility, no spores, and no long chains. Gram stain
is positive.

Gelatine colony.—Colonies are small points, rather opaque, not characteristic.
They are almost wholly under the surface, and never typically grow on the sur-
face. In /ztmus gelatine they are rather dense, strongly acid, and frequently,
though not always, surrounded by minute, irregular spines on the edge. This
type of colony can usually be detected with a little experience, and is the most
characteristic feature of the type. ;

Gelatine stab.—A granular or linear needle growth, and no surface growth.

Agar streak.—TVhere is no growth, or one that is scarcely visible. On milk
agar it grows rather better, but at best it is very scanty.

CLASSIFICATION OF DAIRY BACTERIA. 135

Fermentation tubes.—All three sugar bouillons are rendered acid and there is’
commonly a closed arm growth, but never any gas.

Bouillon.—F requently there is no sign of growth, but there is commonly a
slight sediment.

Milk,—Milk is rendered strongly acid and promptly curdled in from six hours
to two days. The curd is smooth and hard, without gas bubbles, and never
shows any digestion.

Potato,—Usually no growth, but sometimes a thin, transparent film.

Grows at 20° and 37°, but better at 20°. Facultative anaerobic, growing
better without oxygen, and hence curdling milk at the bottom first.

The most characteristic features of this organism are the peculiar Z¢mus-
gelatine colonies, the absence of surface growth, and the smooth, hard, acid curd
in milk.

Variety A.—This differs from the common type simply in its extremely
minute colony, which is invisible to the naked eye, is merely transparent and
does not show the characteristic spines. In our previous list this was called
No. 202, but we now think that it is only a less robust form of the type. In all
other respects the two agree perfectly. The difference of the colonies on gela-
tine, however, is usually very striking, that of the variety A being not more
than I-10 the size of the type. It is less frequently found, also, than that of the
type.

Both the type and variety A show great variability in their acid-producing
power. Sometimes they curdle milk in as short a time as six hours (at 37°),
other cultures in twenty-four hours, others, again, in two or three days, and
finally, some, identical in other respects, fail to curdle it at all, although they
make it strongly acid. These variations in acid-producing power do not seem
to us to be sufficient to warrant us in recognizing them even as varieties. They
are certainly subject to modification in the same cultures. Cultivation in milk
noticeably increases this power. Cultures, when first isolated, show « weak
growth and a weak curdling power, but after a few days’ growth in milk this
power is very greatly increased. A gelatine plate made from fresh milk shows
weak colonies, growing slowly, with a weak acid production. A plate made
from the same milk after two days shows not only more numerous colonies but
colonies much larger, more acid, and growing much faster. It appears thus that
this organism adapts itself to milk, which seems to be its most favorite medium
for growth. : ,

This is the organism which, in previous papers (Annual Rep. Storrs Exp.
Sta., 1901), we have shown to outgrow all other bacteria in milk at 20°, and com-
monly to comprise 99% or more of the bacteria in milk kept at 20°. We have
found this organism all over the United States from the Pacific to the Atlantic,
and it has been sent to us from Europe as one of the most common lactic bac-
teria there. It is present in practically every cheese which we have studied,
including both hard and soft cheeses. It has been many times studied and
been given very many different names. It appears to be the same organism
which has received the following names by different authors; Lactic bacterium
of Kozai, Lactic bacterium of Utz, Strept. acidi lactici of Marpmann, Bact. lactis
acidi Leichmann, B. Jactis acidi Giinther and Thierfelder, B. actdi paralactis a
of Freudenreich (also several others of his cultures are nearly identical), Bac¢ddas

136 STORRS AGRICULTURAL EXPERIMENT STATION.

‘acidilactici Esten. Of these various names that of Leichmann, Bac¢, acidi
lactict, is preferable. The name given by Gtinther and Thierfelder has the
priority, but since this organism is not a bacillus, that name cannot be retained.

Variety B.—This shows a dense, coarse, granular surface colony. It is
very small, only .4u-.74. It does not curdle milk, though it makes it acid and
gives it an astringent taste. It was isolated by Harrison from astringent milk.

Variety C.—This differs from the type in the following points: The colony
shows a surface growth, and there is also a surface growth in gelatine stab.
There is abundant growth in bouillon, with a pellicle. This variety is evidently
more aerobic than the type. One culture of this, from Camembert cheese,
shows a capsule.

Variety D.—This variety never curdles milk though it makes it acid. It
does not stain by the Gram method. It has a thick lobate or moruloid surface
colony, or sometimes smooth. There is a spreading surface on gelatine stab
and a scanty opaque growth on agar streak. Its acid production is feeble and
there is no acidity in saccharose bouillon. In ordinary bouillon there is a sedi-
ment, turbidity, and usually a pellicle. Its growth on potato is scanty. We
have found this several times in different cheeses, and it has also been isolated
from all milk. A@tlch bacterium J., of Koning seems to belong to this type.
(Milchw. Zent. II. 1906, p. 316.)

At this place should be mentioned a series of milk organisms which are
called ‘‘acid fast,” 7.¢., they are not decolorized, by NNO, after being stained
with carbol fuchsin. In this respect they agree with the tuberculosis bacillus,
and hence may sometimes be confused with them in an ordinary microscopic
study of milk. For this reason they are of some considerable significance. At
least nine of them have been described by different authors as follows: JZ.
phlei, Mist bacillus, Grass bacillus, No. 2, and Milk bacillus, all described by
Moeller; the Butter Bacillus of Grassberger; the Butter Bacillus of Binot; the
Butter Bacillus of Rabinowitsch; the Butter Bacillus of Coggi; the Bacillus
Freiburgensis of Koon; also B. Freibengensis, No. 2, and Butter bacilli, Nos.
4, 2,3, gand 5, of Tobler. We have not had an opportunity of studying any
of these, and the descriptions given of them are altogether too inadequate for
classification according to our scheme. Some of them are acid producers and
others are not. Some are white, others orange-yellow or reddish. They form
a miscellaneous lot of bacteria whose relations can not be determined by the
characters given. We find it quite impossible to place them in our scheme at
the present time. It is quite possible that some of the bacteria which we have
described are also ** acid fast,” for we have not used this method of staining in

our routine tests.
°

THE GENUS BACTERIUM, LIQUEFYING,
I. No acid in dextrose or other sugars.

Bact, lactis chromatumn.s. A lemon-yellow, spore-bearing Bacterium.

Morphology.—Size, 34x 1.54. Chains are formed and spores produced, but
there is no capsule.

Gelatine colony.—A liquefying pit, full of threads. There is a central nucleus
with coarse granular masses around it.

CLASSIFICATION OF DAIRY BACTERIA. 137

Gelatine stab.—A deep, dry pit is first formed which, after several days, shows
liquefaction.

Agar streak.—A luxuriant, moist, yellow to white growth.

Bouillon.—A sediment, turbidity, and pellicle. The liquid later becomes
clear.

Milk.—Becomes alkaline and curdles. It subsequently digests into a clear
liquid, with a tenacious scum.

Potato,—Luxuriant, dry, rough, wrinkled, with a brilliant yellow color.

Grows at both 20° and 37°. Aerobic.

Bact. lactis arborescens II, An arborescent spore-bearing Bacterium. This
organism, originally found in 1896, has appeared once or twice subsequently,
although the later organisms differed slightly from the original. One-culture,
sent by Weigmann, differed in points inclosed in brackets.

Morphology.—Rods with square ends. Size, 2u-4ux Iu-1.8m¢, forming long
chains. Spores are produced but no capsules. Gram stain negative.

Gelatine colony.—A very peculiar felted mass of fibers extending through the
gelatine, and a ground glass-like felted surface, on a liquefying disk. The
appearance is variable, but the fibers are characteristic. [Myceloid.]

Gelatine stab.—An arborescent needle growth, liquefying slowly, infundibuli-
form, with a folded, ground glass-like scum.

Agar streak.—Widely spreading, filamentous and somewhat cotton-like on
the surface and extending into the agar; luxuriant, wrinkled, dull. [Cretaceous. ]

Fermentation tubes.—[Dextrose and saccharose acid, lactose not acid, no gas,
closed arm growth in lactose and saccharose. ]

Bouillon.—A flaky turbidity, a sediment, and a ground glass-like scum.

Milk.—Alkaline, curdled and digested. After digestion it may be amber-
colored or colorless. One culture showed a ground glass-like scum.

Potato.—A luxuriant growth, with a white cotton-like surface, extending below
into the potato.

Grows at both 20° and 37°. Aerobic.

In the specimens found later and regarded as the same as the above the
arborescent needle growth was not always found, and the scum on the milk was
lacking.

Bact. lactis filiformis, formerly described, is very similar to the last and
may be the same. It has not been found since the original description. It
differs only in the following points: No arborescent growth in gelatine. The
ground glass-like appearance is lacking. The agar streak shows a dry, white,
lobate growth. Bouillon shows a scum of tangled fibers, and on potato it forms
a thick, slimy growth, yellowish, covering the whole surface of the potato.

Bact. lactis truncatum, A Bacterium with proteus or curled colonies. De-
scribed by us in a previous report and named by Chester. The following is a
description of a culture studied more recently:

Morphology.—Size, 1.24-2.54x .8u-1, forming long chains of square-ended
rods. Spores are produced but no capsules.

Gelatine colony.—An opaque colony, 3{ inch in diameter in two days; curled,
i. ¢., made of twisted threads. In some cases the colony is proteus-like, with
threads in parallel rows.

138 STORRS AGRICULTURAL EXPERIMENT STATION.

Gelatine stab,—Liquefies, stratiform, with a tough, white, mold-like skin,
and, later, with a complete liquefaction and a yellowish scum. The rapidity of
liquefaction is very different in our two cultures.

Agar streak.—A luxuriant, whitish-yellow, rough growth, with an irregular,
lobate or feathery edge. ;

Bouilton.—A tough, felted scum, without turbidity or sediment.

Milk.—Rendered alkaline, curdled in three days, and digested, showing a
thick, folded scum. In twelve days a translucent surface with a curd below.

Potato.—Luxuriant, white, velvety or powdery dry growth.

Grows at 20° and 37°. Aerobic.

\

Bact. lactis Michiganii n. s. A white, spore-bearing Bacterium. This
organism, sent by Marshall, does not agree with any that we have found,
although closely allied to the last. The most striking difference is in the colony,
but this may be due to the greater vigor of liquefaction. At present, however,
we keep it as distinct.

Morphology.—Size, 1.84x.94, forming chains. Spores developed. Gram
stain negative.

Gelatine Colony.—A rapidly liqwefying colony, with a uniformly cloudy
liquid.

Gelatine stab.—Needle growth beginning to liquefy in one day, infundibuli-
form; liquefaction complete in three days.

Agar streak,—Spreading widely, moderately thick, wrinkled, opaque, white,
not luxuriant. ,

Fermentation tubes.—No acidity, no gas, no closed arm growth.

Bouillon.—A sediment, turbidity, and a wrinkled pellicle.

Milk.—Alkaline, curdled and completely digested. At 37° slightly pinkish,
but not at 20°.

Potato. — Luxuriant, filiform, thick, alveolate, opaque, gray-brown, dry,
wrinkled. g

Grows better at 37” than at 20°. Aerobic.

Bact. lactis Genevum ns. A white, spore-bearing Bacterium, sent us by
Harding.

Morphology.—Size, 34-84 x 1.44. No chains. Spores formed chiefly at the
ends of the rods. Rods with square ends. Gram stain positive. (One culture
negative.)

Gelatine Colony.—A rapidly liquefying colony that may be a smooth liquid
mass or may be cloudy. On “tmus gelatine there may be a wrinkled film.
Colony not acid.

Gelatine stah.—A needle growth and a stratiform liquefaction, beginning in
one to three days.

Agar streak.—Spreading, thin or raised, smooth, whitish or creamy, moist or
sometimes dull, luxuriant.

Fermentation tubes.—No acidity or gas in any sugar bouillon; closed arm
growth usually seen. (A slight acidity may appear for a day or two in dextrose,
but it then disappears.)

Bouillon.—A sediment, turbidity and pellicle.

CLASSIFICATION OF DAIRY BACTERIA, 139

Mitk,—Is rendered alkaline, curdled and digested completely, with a promi-
nent odor.

Potato.—Spreading, thin or raised, smooth, opaque, cream colored, luxuriant;
discolored.

Grows at 20° and 37°. Facultative anaerobic.

Bact. lactis erythrogenes Grotenfeld. Bacterium with pink fluorescence. We
have several times found bacteria that belong probably to this well known type.
None of them produce much red color in milk, although some render it of a
pinkish color, and one turned it deep red after several weeks’ growth, All of
them, except variety D, produce a peculiar pinkish fluorescence in agar. We
give below the characters of gne of the cultures isolated, with the others as
varieties.

Morphology.—Size, 1.24x.9u-Iu. Not forming chains. It forms no spores,
stains by the Gram method and shows a capsule.

Gelatine colony.—Round, raised, smooth, homogeneous, entire, translucent,
yellowish, shining, later liquefying.

Gelatine stab.—Begins to liquefy in three days, stratiform.

Agar streak,—Filiform, raised, smooth, translucent, flesh color or pink,
shining, luxuriant. The agar shows a pink fluorescence.

Fermentation tubes.—Dextrose and saccharose show growth in closed arm,
but not lactose. No acidity nor gas produced in any sugar bouillon.

Bouillon.—A flocculent sediment, a membranous pellicle, and a decided tur-
bidity.

Milk.—No change in reaction. The milk may curdle in ten days and begin
to digest. The digestion is nearly complete in three weeks and the liquid is of
a pinkish color, with a slight odor.

Potato.—A luxuriant growth, white, with discolored potato. There is no
pink color shown. ‘

Grows both at 20° and 37° _—‘ Facultative anaerobic.

This organism was from a yellow slime on the surface of cheese.

Variety A.—Differs from the above in showing no closed arm growth, a
yellowish growth on agar, no curd or digestion in milk, and a scanty growth on
potato.

Variety B.—Shows yellow growth on agar. No pellicle on bouillon. Milk
becomes alkaline and digests into a red liquid, which later becomes very red.
Growth on potato scanty.

Variety C.—Size, .34 x .5¢-.6u. No pellicle on bouillon. Milk digests;
pinkish. Potato scanty. Growth on agar not yellow.

Variety D.—This fails to produce a pink fluorescence, but has a yellow
growth and is wrinkled. The milk is rendered pink, and the growth on the
potato is yellow and scanty. This variety is, perhaps, Bact. erythrogenes of
Dyar (Trans. N. Y. Acad, of Sci., 1895). Probably the same as Bact. luteum
of Zimmermann. (See p. 142.)

I40 STORRS AGRICULTURAL EXPERIMENT STATION.

Bact, lactis rubrum. Non spore-bearing, pink Bacterium, This has not been
found since its original description in 1899. Its characters, as then given, are
as follows:

Morphology.—Size, 2u-44x.gu. Forming chains. No spores, no capsule.

Gelatine colony.—A bead-formed colony, .7 mm. in diameter, with a granular
edge. Liquefies with a nucleus and a clear zone.

Gelatine stab. A slow liquefier, stratiform, producing a clear liquid with a
scum and a sediment.

Agar streak.—Luxuriant, wrinkled, dull orange-yellow or pinkish.

Bouillon.—A sediment, but no pellicle nor turbidity. :

Milk.—Becomes alkaline and curdles after several days at 37°. It digests
into a dirty liquid.

Potato,—Glistening, smooth, pink or salmon-qolored, luxuriant.

Grows at both 20° and 37°.

Variety A.—Found later. Agrees with the above, except that it is of an
orange rather than a pink color.

Bact. lactis Burrin. >. A reddish, bitter milk organism, described by Burr
and Dugelli. As described by them its characters are as follows: (Cent. f.
Bac, II., XV., p. 709.)

Morphology.—Size, tu-34x.74. Nochains; no spores. Gram stain nega-
tive.

Gelatine colony.—Surface colony in a liquefying area, 4g mm. in diameter.
In fourteen days it is 4% mm. in diameter, of a clear brown color.

Gelatine stab.—Begins to liquefy,in four days; infundibuliform.

Agar streak.—Luxuriant, smooth, lobed, reddish.

Fermentation tubes.—No acidity, gas, nor closed arm growth.

Bouillon.—A turbidity, but no sediment nor pellicle.

Adilk.— Becomes acid, but does not curdle or digest. It becomes a rusty
red, with a cheesy smell and, later, a bitter taste.

Potato.—No growth.

Grows at 20° but not at 37°. Aerobic.

Bact. lactis citronisn. s. A non spore-bearing, lemon-yellow Bacterium.

Morphology.—Size, 1x .6u, forming chains. No spores are produced, and
no capsules.

Gelatine colony.—Small pits are produced, with a nucleus and a lighter outer
zone, which may be variously streaked.

Gelatine stab.—A slow liquefier, producing a cratiform liquefaction, with a
dense sediment and a yellow liquid. At first clear, but later cloudy.

Agar streak.—A luxuriant, thick, folded growth, at first greenish-yellow, and
later lemon-yellow.

Bouillon.—A turbidity and a sediment.

Milk.—Becomes alkaline and digested into an amber-colored or pale yellow
liquid. Sometimes it curdles before digestion.

Potato.—Thick, smooth, flesh-colored, and later lemon-yellow; or sometimes
lemon-yellow from the start.

Grows at both 20° and 37°. Aerobic.

CLASSIFICATION OF DAIRY BACTERIA. I41

Variety A.—This variety, also found in milk in Middletown, agrees with
the above in most respects, differing in the following points: Size, 34x 1.5m.
Spores are produced. The gelatine forms a pit full of threads. Milk is curdled
and after digestion shows a clear liquid and tenacious scum. The potato
growth is wrinkled. ,

Variety B.—This organism forms no chains nor spores. Its colony is lemon.
yeilow, even when viewed under the microscope, a very unusual character; later
it liquefies and sends processes into the gelatine. Its gelatine stab is infundib-
uliform, or a dry pit. Its growth on agar is lemon-yellow and very thin.

It is doubtful whether these three varieties should be grouped together, and
certainly they should not if it is true that two of them do not form spores. We
have lost these cultures and are unable to verify the characters above given.
Therefore we insert them in their original form.

Bact. lactis minutissimum., A very slender, orange Bacterium, We have
not fouud this since the original description in 1899. The following are the
characters as then described.

Morphology.—Size, 1.5@x.4¢. Forming long chains, and not producing
spores,

Gelatine colony.—Surface colony irregulary branching; ‘deep colonies burr-
like, with a yellow centre and irregular processes, or sometimes simply lobed.
Rays extend from a liquefying pit into the gelatine.

Gelatine stab,—Begins to liquefy in two days with an infundibuliform or crati-
form liquefaction, and a brilliant yellow sediment.

Agar streak.—A luxuriant, widely spreading, orange growth, covering the
whole surface.

Bouillon.—A sediment, turbidity and a pellicle.

Milk,—Rendered alkaline and becomes somewhat thick and dark colored, but
no visible digestion.

Potato.—Luxuriant, deeply orange.
Grows better at 20° than at 37°. Aerobic.

Variety A.—A second culture, found later, may be a variety of the same,
although it is‘not so small. It is .6% in diameter, and does not appear to form
chains. Its colony is brilliant yellow, smooth and translucent, but does not
show the irregular processes. Milk does not become thick. The color on
potato is bright yellow rather than orange.

Bact. lactis Marshalli u.>. A slimy milk, yellow Bacterium. This is the
organism with which Marshall has worked and which was shown by him to
have a hastening action upon lactis bacteria (Cent. f. Bact. II.). The follow-
ing characters were determined by us from cultures sent by him, and agree
_essentially with those determined by him:

Morphology.—A rod, not forming chains. Size, 1.24x.3u. (Marshall’s

measurements, I.7#-5.25 x .8u-.875u4.) It produces no spores, has no capsules
and Gram stain is negative.

I42 STORRS AGRICULTURAL EXPERIMENT STATION.

Gelatine colony.—A slowly liquefying, granular colony, which may later
become large, irregular and slimy.

Gelatine stab.—Begins to liquefy in two to three days, infundibuliform. In
nine days it liquefies &% inch. Upon Atmus gelatine it forms a rough, red-
brown colony, with a dark center. Not acid.

Agar streak.—A luxuriant, viscous colony, filiform, raised, smooth, cream-
white or gray, but later lemon color.

Fermentation tubes.—No acidity, gas, nor closed arm growth in any bouillon.

Bouillon.—A sediment and turbidity, and a pellicle formed around the edges
of the tube.

Milk.—Becomes alkaline and digests, but does not normally curdle. Has a
prominent odor, and is slimy.

Potato.—A luxuriant, filiform, effused, smooth, lemon-yellow growth.

Grows both at 20° and 37°. Aerobic.

We have found essentially the same organisms in Middletown, and Harding
has sent us one from New York which failed to digest milk, but agreed in other
respects. This latter one may, perhaps, be called variety A.

Variety B.—An organism isolated from Camembert cheese, sent us direct
from France, really belongs here, although differing in the following charac-
ters: Its colony is white. It is not viscous. It forms no pellicle. It does
not digest milk. Its growth on potato is orange. We have found the same on
Camembert cheese from New York markets. :

Bact. lactis Limburgiin. s. A non spore-bearing, orange-yellow Bacterium,
This organism was isolated from milk and described by Burri and Dugelli as
follows. (The name is our own):

Morphology.—Size, 1.5u-3a@x .5¢. Forming no chains and no spores. .

Gelatine colony.—A round, brownish colony, 1 mm. in diameter. After six
days a yellow disk is seen in its cloudy liquid.

Gelatine stab.—A needle growth and a surface growth, but after six days
liquefaction begins; stratiform. :

Agar streak,—Luxuriant, smooth, glistening, dirty yellow.

Fermentation tubes.—Not described, but no acidity is produced, and probably
no gas in sugar bouillons.

Bouillon.—Liquid becomes turbid, but shows no pellicle.

Milk.—No change in reaction, and no curdling. The milk becomes digested
and has a Limburger smell.

Potato.—Scanty yellow, glistening. Potato not discolored. '

Bact, lactis luteum, This name was given by Zimmerman (Cent. f. Bac. II.,
XI., p. 200,) to a type isolated by him from the udder. Several of our cultures
agree with it as closely as can be determined from his incomplete descriptions.
It agrees closely with Bact. erythrogenes, except in lacking a pink fluorescence,
and in not making milk red or pink. The characters given below are from our
own cultures, to which we have given Zimmerman’s name. We have found the
organism in Camembert cheese from the markets, probably imported from
France.

CLASSIFICATION OF DAIRY BACTERIA. 143

Morphology.—A rod, formingno chains. Size, 1.24x.84. No spores no
capsules. Gram stain positive.

Gelatine colony.—A slowly liquefying colony, with a dense central growth
and aclear liquid.

Gelatine stab,— A slow liquefaction; crateriform.

Agar streak,—A luxuriant, filiform, raised, rugose, lemon-yellow growth;
dull and wrinkled.

Fermentation tubes,—No gas, no acidity, and no closed arm growth in any

, bouillon,

Boutllon.—A sediment, membranous pellicle and a slight turbidity.

Milk.—Becomes alkaline, but no other visible change.

Potato.—A luxuriant, spreading, thick, contoured growth, opaque, white or
brownish-yellow. :

Grows at 20° and 37°. Aerobic.

The color of this organism varies from a brilliant yellow to a dull yellow.
Some cultures do not appear to grow on potato and show a slight digestion of
milk.

Variety A.—One culture we regard as distinct enough to be recognized as a
variety. It was obtained from milk rather thanfromcheese. Its size is 84x .34,
and it does not stain with Gram. Its colony is round, moderately thick, smooth
and yellow, and then forming a pit. Upon Xtmus gelatine it is pale and thin.
It produces no pellicle on bouillon, but curdles and digests milk, turning it
slightly yellow. Its growth on potato is dry, luxuriant and lemon-yellow.

Bact, lactis Ashtoniin. s. A non spore-bearing, yellow Bacterium.

Morphology.—Size, 1.2u-3.5@x 1.24. No chains, no spores nor capsules.
Gram stain irregular.

Gelatine colony.—A slow, liquefying pit, with a cloudy liquid, tinged with
yellow, Atmus gelatine is not acid.

Gelatine stab.—A needle growth and a napiform liquefaction, beginning in
three days.

Agar streak.—Filiform, raised, smooth, yellow, moist, viscous.

Fermentation tubes.—No acidity or gas in any bouillon, closed arm growth in
all. (Saccharose is slightly acid after several days.)

Bouillon.—Sediment, turbidity and pellicle.

Milk.—Is rendered alkaline, curdled and digested completely, with a yellowish
surface and a strong odor.

Potato.—Filiform raised, smooth, yellow, luxuriant; no discoloration.

Grows at 20° better than at 37°. Facultative anerobic,

Bact, lactis album n. >. A white, liquefying Bacterium, not acid. Found in
the udder at Storrs. It may be the same as Bact. /uteum, without pigment. Its
characters are as follows: :

Morphology.—Rods, forming no chains. Size, Iu-3ux .7u-.gu. It forms no
spores, no capsules, and Gram stain is positive.

144 STORRS AGRICULTURAL EXPERIMENT STATION.

Gelatine colony.—A very slowly liquefying colony, not characteristic; some-
times the gelatine dries before liquefaction takes place.

Gelatine stab.—Begins to liquefy in three days, and is about 1¢ liquefied in
twenty-one days. Napiform and stratiform.

Agar streak.—Filiform, raised, smooth, opaque, cream-white, shining, luxuri-
ant, viscous.

Fermentation tubes,—No acidity, gas, nor closéd arm growth.

Bouillon.—A sediment, turbidity and pellicle.

Milk.—Becomes alkaline, is curdled, or sometimes digests without curdling.
The digested milk is very slimy and of a slight straw color (in one culture, pink).

Potato.—A very abundant, spreading, convex, smooth, brown growth; potato
discolored.

Grows at 20° and 37° abundantly. Aerobic.

Variety A.—An organism found in the pink slime on Camembert cheese
closely resembles this, but differs in the following: It is not viscous on agar,
nor does it make milk slimy. It produces no pellicle in bouillon, and neither
curdles nor digests milk.

II. Acid in dextrose or other sugars.

Bact, lactis musci. An arborescent bacterium with myceloid colonies. This
was found in milk in Middletown, in cheese, and one culture was sent from
Kiel isolated from Mazoon. Perhaps identical with Bact, mycoides (Fligge).

Morphology.—Long filaments made up of rods 34x1. Central spores no
larger than the rods. Gram stain positive.

Gelatine colony.—Myceloid, branching, radiating colonies before liquefaction.
Commonly forming a thin, white, velvety, or ground-glass surface upon the
liquefying gelatine. Litmus gelatine is not acid.

Gelatine stab,—There is at first an arborescent needle growth, but liquefaction
begins in one day, and a wrinkled glass-like surface is formed.

Agar streak,—Luxuriant, thin, white, wrinkled. In one culture threads grow
down into the agar, and the agar turns dark colored.

Fermentation tubes,—Growth is somewhat variable. In one culture dextrose
is rendered acid with a closed arm growth in all sugar bouillons. In another
culture dextrose and saccharose are acid, and there is no closed arm growth.
No gas.

Bouillon.—A turbidity, sediment and pellicle, except in one culture where
the pellicle is wanting.

Milk.—Curdles with amphoteric reaction and subsequently completely di-
gested.

Potato.—Luxuriant, thin, whitish, sometimes wrinkled; potato may be dis-
colored.

Grows at 20° and 37°. Aerobic.

Variety A.—One culture was sufficiently different to be separately tabulated.
The differences are as follows: Size, 2.5¢x1.4u. Gram stain negative.
The gelatine colony shows masses of bacteria hanging together. They are
arborescent under mica. A dry pit formed in gelatine stab. Agar growth not
wrinkled. No closed arm growth. The reaction of milk is alkaline. Here
belong Nos. 14 and 16 of Adametz.

CLASSIFICATION OF DAIRY BACTERIA. 145

Bact, lactis cretaceum n. >. A non-arborescent, spore-bearing Bacterium.

Morphology.—Size, 3-5" x 1.4u. Nochains nor capsules. The Gram stain
is positive. Spores are produced, no larger than the rods.

Gelatine colony.—Not characteristic; a rather slowly liquefying pit forms,
without any distinguishing marks. In /¢mus gelatine it is not acid.

Gelatine stab.—No needle growth. In one day it begins to liquefy; strati-
form or sometimes infundibuliform.

Agar streak.—Filiform, raised, smooth, cretaceous, white or flesh-colored,
moist, luxuriant.

Fermentation tubes.—Acidity and closed arm growth in dextrose and saccha-
rose; closed arm growth, but not acid inlactose. No gas.

Bouillon.—A sediment, a slight turbidity and, later, a slight pellicle.

Milk.-—Becomes alkaline, curdles promptly and completely digests, with a
_ Prominent odor.

Potato.—Spreading, raised, smooth, cretaceous, white, luxuriant; potato dis-
colored.

Grows at 20° and 37°. Facultative anaerobic.

Bact, lactis lobatum n.s. An orange, acid liquefier. This organism was
found in stable dust and in milk direct from the udder. The two cultures
differ slightly in color and bouillon growth.

Morphology.—Size, .84-1m x .54. No chains, no spores, no capsules. Gram
stain positive.

Gelatine colony.—A round, raised, smooth homogeneous coijony, sometimes
yellowish, with a lobate center and a clear ring when liquefying. Litmus gelatine
shows an acid, liquefying colony.

Gelatine stab,—Slow liquefier, at first saccate, and then stratiform. The
liquid is cloudy.

Agar streak.—A smooth, raised, thin, orange-colored colony, luxurious.

Fermentation tubes.—All three sugars are rendered acid, and there is growth
in the closed arm, but no gas.

Bouillon.—A sediment, turbidity, and pellicle. In one culture there is no
pellicle.

Milk.—Rendered acid, but not curdled. It is digested and turned amber-
color or reddish, with a jelly-like sediment.

Potato.—Thick, opaque, orange-colored, luxurious.

Grows well at 20°, hardly at all at 37° Facultative anaerobic.

Bact. lactis cloacae n.s. This organism appears much like B. cloacae of
Jordan, differing slightly in its growth in bouillon and on potato. Its lique-
fying power is also less. It was found only once in milk, in Middletown.

Morphology,—Size, .7#-.8h X 5M. No chains, no spores. Gram stain nega-
tive. It shows a capsule,

146 STORRS AGRICULTURAL EXPERIMENT STATION.

Gelatine colony,—A round, thick, smooth, homogeneous colony, I mm. in
diameter; white, acid on Atmus gelatine. :

Gelatine stab.—A dry pit is first formed, which later liquefies, infundibuli-
form.

Agar streak.—A narrow, raised, smooth growth; opaque, whitish, dull,
fairly luxuriant. ‘

Fermentation tubes.—Acidity, closed arm growth and gas in all sugars.
Bouillon.—A sediment and turbidity, but no pellicle.

Milk.—Acid and curdled, showing a clear whey, but there is no visible
digestion. A sour odor.

Potato.—A scanty, white growth, with a discolored potato.
Grows at 20°, and moderately at 37°. Aerobic.

Bact, lactis liguaerogenes n. ». A gas-producing, liguefying Bacterium.

Morphology.—Size, 1u-1.6" x .74. No chains, no spores, and Gram stain
negative.

Gelatine colony.—A non-characteristic, rapidly-liquefying colony.

Gelatine siab.—Begins to liquefy in two days; complete in nine days.

Agar streak,—Not luxuriant, spreading, thin, smooth, white.

Fermentation tubes.— Dextrose and saccharose show acidity, gas and closed
arm growth. Lactose shows closed arm growth but no acidity or gas.

Bouillon.—A turbidity, sediment and a pellicle.

Milk.—Is curdled without change of reaction, and is digested with an odor
of Brie cheese.

Potato.—Not luxuriant, spreading, smooth, thin, white; potato discolored.

Grows at 20° and 37°. Facultative anaerobic.

.

Bact. visco fucatum Harrison and Barlow. A slimy milk Bacterium with
blue pigment. This organism, isolated and described by Harrison and Barlow,
from oily butter, seems to be new. (Trans. R. S. of Canada, XI., 1905-6.) It
was not the cause of the oiliness of the butter, but is peculiar in showing a wide
range of color. Its characteristics are given below in condensed form.

Morphology.—Size, 1¢-1.8p. x .6u-.94. No long chains, no spores. Gram
stain positive, and an evident capsule. Branched involution forms in all cultures.

Gelatine colony.—A slimy colony, yellowish-green, with crystals in the gela-
tine. When carbohydrates are present, the gelatine is blue or green.

Gelatine stab,—Liquefies in ten days, complete in two months; cratiform.
The liquid is of a dark inky color above and rusty below.

Agar streak.—A slow, smooth growth, viscous, no pigment.

Fermentation tubes.—Not stated, but probably acid without gas.

Bouillon.—A sediment and turbidity, but no pellicle. The liquid is alkaline,
slimy.

Milk.—Rendered acid and curdled after several days; later digested, with a
greenish-blue color, and becomes slimy.

CLASSIFICATION OF DAIRY BACTERIA. 147

Potato,—Yellowish-white, slimy, luxuriant. The potato becomes light-blue
and, later, amber-colored.

Grows at both 20° and 37°. Aerobic.

Bact, lactis brevis nr. >. A white liquefier. This has been found several
times in milk and cheese here and elsewhere. It seems to be identical with one
sent by Freudenreich and is apparently a widely distributed species.

" Morphology.—Size, .7u-.9u x .54-.64. No chains, no spores. Gram stain
irregular; no capsule. ‘

Gelatine colony.—A round, thin colony, lobed, whitish. It may show acidity
in tmus gelatine, and it slowly liquefies.

Gelatine stab.— Begins to liquefy in from one to two days, and may be com-
pletely liquefied in from three to twelve days. Stratiform.

Agar streak.—Fairly luxuriant, smooth, white, not characteristic.

Fermentation tubes.—All thrée sugars are acid, and there is usually growth
in the closed arm, but no gas. .

. Bouillon.—A sediment, but no turbidity, and no pellicle.

Milk.—Rendered acid and curdled, and later, is partly digested.

Potato.—Barely visible; thin and white. ‘

Grows at 20° and 37°. Facultative anaerobic.

Found many times, the numerous varieties showing some differences. The
Freudenreich organism is longer, Iu in length. It makes milk acid and bitter.
This is apparently A/ilch. Bacterium LI. of Koning (Milchw. Zent. II., p. 317,
1906).

Variety A.—This organism, sent by Harding, differs in the following points:
Size, 2-34x 1.2. Liquefaction of gelatine tube never complete. A pellicle in
bouillon. Rather more luxuriant on potato. . :

Bact. lactis fluorescens u. >». A fluorescent Bacterium, This organism,
being non-motile, seems to be different from the common fluorescent form. We
have found it but once, in New York city milk. It may be only a non-motile
variety of B. fluorescens, but we have been unable to detect any motility in our
cultures.

Morphology.—Size, I.4u-2.5m xX u-.gp. No chains, no spores, no Gram
stain and no capsule.

Gelatine colony.—A slow liquefier, forming a peculiar lace-like colony in a
pit, with a dense centre. :

Gelatine stab.—A needle growth, stratiform; liquefaction beginning in one
day.

Agar streak.—Filiform, capitate, smooth, translucent, white, luxuriant, with
a green fluorescence. ,

Fermentation tubes,—Dextrose and saccharose are rendered acid; lactose is
not; no closed arm growth; no gas.

Bouillon.—A sediment, turbidity and a pellicle.

148 STORRS AGRICULTURAL EXPERIMENT STATION.

Miik.—Rendered alkaline and curdled at 20°, but not at 37°. Digestion
occurs later and the milk has a strong odor.

Potato.—A filiform, raised, smooth, white, luxuriant growth.

Grows at 20° and only slightly at 37°. Aerobic.

Bact. lactis plicatumn.s. A non-acid, white, liquefying Bacterium. Isolated
from milk by Harding.

Morphology.—Size, 3u-5u x .8u-.94, growing in long chains. No spores, no
capsules, and Gram stain irregular.

Gelatine colony.—A slow liquefying colony, showing a peculiar folding.

Gelatine stab,—A needle growth, beginning’to liquefy in one day, and about
half liquefied in ten days; infundibuliform.

Agar streak.—Filiform, thick, smooth, opaque, whitish, moist, moderately
luxuriant.

Fermentation tubes.—All three sugar bouillons are slightly acid, but there is
no closed arm growth, and no gas.

Bouillon.—A sediment and slight turbidity, but no pellicle.

Milk.—Rendered alkaline, curdled in three days and subsequently digested.

Potato.—Spreading, thick, mottled, wrinkled, white, luxuriant, with dis-
colored potato.

Grows at 20° and. 37°. Aerobic.

Variety A.—Differs in the following points: No chains observed. Colony

without the peculiar folding. Digestion of milk not noticeable. Growth on
potato scanty and not wrinkled.

Bact, lactis Gorinii n. 8. A non-acid, white, liquefying Bacterium. The
original of this organism was sent from Italy by Gorini. Two very similar cul-
tures were found by us, one in New York and one in Connecticut. The
original culture had the following characters:

Morphology.—Size, 1.54-2.54 x Iu. Rods with square ends. No chains, no
spores. Gram stain positive.

Gelatine colony.—A slow liquefier, producing a large pit with irregular mottled
clusters of bacteria,

Gelatine stab,—Begins to liquefy in two days and is 3{ complete in three
weeks. Infundibuliform.

Agar streak.—Spreading, raised, smooth, opaque, white, moist, luxuriant,
viscous.

Fermentation tubes.—Dextrose and saccharose are acid; lactose not acid.
No closed arm growth; no gas.

Bouillon.—A sediment, turbidity, and a pellicle.
Milk.—Made strongly alkaline, curdled and completely digested at 20°.

Potato,—Spreading, thick, smooth, opaque, moist, luxuriant; potato dis-
colored.

Grows well at 20° and 37°. Aerobic.

CLASSIFICATION OF DAIRY BACTERIA. 149

Variety A.—Isolated from milk in Connecticut. Is not quite so large. Gram
stain negative. Saccharose is not acid. Produces iridescent brown color.
Does not grow well at 20°. Milk has a peculiar color.

Variety B.—(From milk in New York city.) Saccharose is acid. There is
no pellicle on bouillon. No digestion of milk. Growth on potato is scanty.

Variety C.—(From Brie cheese.) Has a simple, granular, rapidly-liquefying
colony. Shows growth in closed arm of fermentation tubes, and a pellicle on
bouillon, The milk develops the odor of Brie cheese.

Varieties A and B are probably cultures of the same organism, one of
which is more vigorous than the other. Here evidently belongs Bact. C. of
Miller (Arch. f. Hyg. LXVII., p. 127), which agrees in all points, but shows a
blue fluorescence in gelatine, and gas in dextrose bouillon.

Bact. lactis magnum n.s. A non-acid, white, hguefying Bacterium,

Morphology.—Size, 34x 1.5. Chains produced; no spores or capsules.
Gram stain positive, Rods with square ends.

Gelatine colony.—A fairly rapidly liquefying pit, which may be filamentous
and with ciliated edge. On ditmus gelatine it is not acid.

Gelatine Stab.—A needle growth, which may be arborescent and later lique-
fying; stratiform. Liquefaction begins in one to three days; complete in three
weeks.

Agar streak.—Filiform or spreading, thick, punctate, opaque, white, moist,
luxuriant.

Fermentation tubes.—Acid in dextrose only. No closed arm growth and no

gas.
Bouillon.—A sediment, a turbidity, and a pellicle.

Milk.—Becomes alkaline, curdles after three days and digests into a brownish
liquid, and a prominent odor.
Potato.—Spreading, thick, contoured, translucent, white; potato discolored.

Grows at 20° and 37°. Aerobic.

Bact. lactis flocculus u.s. An acid, non-curdling, liquefying Bacterium.,
From Camembert cheese.

Morphology.—Size, 1u-2¢ x TM. No chains, no spores, Gram stain positive.

Gelatine colony.—A slowly liquefying colony, which is lobate or moruloid.

Gelatine stab,—A needle growth and a surface growth, which begins to liquefy
in ten days.

Agar streak.—A filiform, raised, smooth, opaque, white colony, rather
scanty.

Fermentation tubes.—Dextrose ts acid, but no other sugar bouillon. No
closed arm growth, no gas. One culture is acid in all sugars.

Bouillon.—A sediment, turbidity, and a pellicle. One culture shows no
growth. :

Ailk.—Acid but not curdled, and shows no digestion. Hasa prominent odor.

150 STORRS AGRICULTURAL EXPERIMENT STATION.

Potato.—Spreading, thin, contoured, white, moist, luxuriant; potato dis-
colored.

Grows better at 20° than at 37° Aerobic.

THE GENUS PSEUDOMONAS.
I. Non-liquefying.

Ps. lactis Middletownii n. s. A Pseudomonas, producing gas, but no acidity,

Morphology.—Size, 1.4.x .8u-.94. No chains, no spores, no capsules.
Gram stain negative.

Gelatine colony.—A round, raised, smooth, gyrose, entire, gray-white colony.

On “litmus gelatine the colony is coarsely granular, looking like a colony of

yeast.

Gelatine stab.—A dry pit is formed, with needle growth.

Agar streak.—Filiform or spreading, smooth, thin, gray-white, moderately
luxuriant, moist.

Fermentation tubes.—No acidity in any sugar bouillon, but closed arm growth
and gas produced in all.

Bouillon.—A sediment, turbidity and a pellicle.

Milk.—Rendered acid and curdled after several days, and subsequently
slightly digested.

Potato,—A \uxuriant, spreading, thick, gray-white growth; potato discolored.

Grows at 20°, but hardly at all at 37°. Aerobic, or facultative anaerobic.

Ps. fluorescens aurea Weigmann. A fluorescent, non-liguefying Pseudo-
monas. This culture was sent me by Weigmann, but ‘had lost its fluorescent
character when received by us. Its characteristics, as given below, were made
in our laboratory. This is perhaps Ps. convexa of Wright. We have several
times found a similar organism in milk, but unfortunately at the time we did
not make a study of their flagella; but we have no doubt that they belong
with this organism of Weigmann. We list one of our own organisms as
variety A.

Morphology.—Size, 2.5@x.9gu. Short chains. There are no spores nor
capsules and the organism does not accept the Gram stain. [Our organisms
were usually shorter. |

Gelatine colony.—A round, raised, contoured, grumose colony, of a brownish-
red color. [A green halo around the colony iscommon.] On Atmus gelatine
it is much folded.

Gelatine stab.—A filiform, needle growth and a raised, surface growth. [The
surface growth is thick, and the gelatine may show a green color. |

Agar streak.—A filiform, smooth, thin growth of a yellowish color, moder-
ately luxuriant. It was probably originally fluorescent, but this character has
been lost. Our own cultures always showed the fluorescence.

Fermentation tubes.—No acidity, gas, nor closed arm growth in any sugar
bouillon.

CLASSIFICATION OF DAIRY BACTERIA. I51

Boutllon,— A red sediment, a ring-like pellicle, and slight turbidity are
formed. [Greenish. ]

Milk.—Rendered slightly alkaline, or there is no change in the reaction.
No curdling nor digestion occurs, and there is no change in color.

Potato.—A filiform, raised, contoured growth, of a brownish-yellow.to an
orange color; luxuriant, and with the potato discolored. [Yellow to brown,
and sometimes folded. ]

Grows at both 20° and 37° ~—- Aerobic.

Variety B.—Isolated from milk here, is a similar, non-liquefying fluorescent
monotrich, which we regard as the same. It is quitecommon and agrees closely
enough with the above to be called by the same name. It commonly makes
the gelatine green, and has a strong fluorescent action on agar. Milk is also
turned green, but is otherwise unchanged. On agar the growth is rather
scanty, but is luxuriant on potatoes, of a brown color to white. On potato it
may be folded.

Ps. lactis Estenii n.s. A Pseudomonas with a smoky fluorescence. This
organism is quite common in milk. Another one practically identical agrees in
all points except that it does not produce the smoky fluorescence,

Morphology.—Size, .8u-1.2" x .4". No chains are formed, no spores nor
capsules, and the organism does not accept the Gram stain.

Gelatine colony.—A round, smooth, capitate, contoured, homogeneous colony,
of a cream-white to a gray color. On “tmus gelatine it forms a pale, thin
surface colony.

Gelatine stab.—A filiform, needle growth, and a raised, dry, not spreading,
surface growth.

Agar streak.—Filiform, raised, smooth growth, translucent; gray in color;
luxuriant and slightly viscous. The agar becomes smoky.

Fermentation tubes.—No acidity, gas, nor closed arm growth in any sugar
bouillon.

Bouillon.—A sediment and flocculent turbidity are formed, but no pellicle.

Milk.—No action on milk.

Potato.—A filiform, thin, smooth growth; gray in color; moderately luxuriant.

Grows both at 20° and 37°. Aerobic.

Variety B differs from A in being slightly larger, 1.84-3¢ x .7#, and in pro-
ducing no fluorescence.

Ps. lactis fliformis n.s. A yellow, non-liquefying Pseudomonas. Found
once in New York milk.

Morphology.—Size, 2.54-3.5é@ x .8u-.gu. No capsule, and no Gram stain.
Spores are produced which are frequently seen as long chains. One long
flagellum.

Gelatine colony.—A round, convex, smooth, entire colony of a creamish or
yellow color, On litmus gelatine the colony is red-brown, non-acid.

152 STORRS AGRICULTURAL EXPERIMENT STATION.

Gelatine stab.—A filiform needle growth, and a flat surface.

Agar streak.—A filiform, raised, smooth, yellowish growth, not very luxuriant,
moist.

Fermentation tubes.—Dextrose and saccharose bouillons are rendered acid.
There is no gas nor growth in the closed arm in any bouillon.

Bouillon,—A sediment, a ring-formed pellicle, and a flocculent turbidity.

Mitk.—Rendered acid, but no other change produced.

Potato.—A beaded, thick, punctate growth, of a yellow color, not very
luxuriant. Potato discolored.

Grows better at 20° than at 37°. Aerobic.

Ps. pseudo tuberculosis Klein. This organism, isolated by Klein from Lon-
don milk, was sufficiently well described by him to be inserted here as follows:

Morphology.—A rod, forming long chains; individual elements I.2u-1.8u x

‘ .4¢-.5@. Gram stain positive. No spores nor capsules described.

Gelatine colony.—A white surface colony, somewhat granular, and resembling
that of B. col. No gas is produced, however.

Agar streak.—Resembles B. coli, but is less luxuriant.

Fermentation tubes.—Not described but, doubtless, no acidity nor gas is
produced.

Bouillon.—A turbidity and a slight pellicle, but no sediment.

Milk.—No action on milk.

Potato.—A thin, crenate, faintly brownish growth, with a whitish-yellow
margin.

This is found in 8% of London milk, and is said to be pathogenic for Guinea
pigs.

Ps. lactis viridis ns. A Pseudomonas, turning gelatine green. This
organism has been found only once, in milk from Colchester.

Morphology.—Size, .gu-I¢ x .4u-.5@. Spores are produced, and there is no

; capsule. There are no chains, and the Gram stain is negative.

Gelatine colony.,—A round, raised, smooth, homogeneous colony, entire, yel-
lowish, moist.

Gelatine stab,—A needle growth with raised surface. The gelatine is turned
green.

Agar streak.—A filiform, raised, smooth, translucent, white growth, quite
luxuriant, moist; no fluorescence.

Fermentation tubes.—Dextrose rendered acid and shows growth in closed
arm. There is no gas. In the other sugar bouillons there is no acidity, gas,
nor closed arm growth.

Bouillon.—A flocculent sediment, a pronounced turbidity, but no pellicle.

Mitk.—Very slightly acid, but no further change.

Potato.—A filiform, thin, smooth, creamish-colored, moist growth; not very
luxuriant. Potato discolored.

Grows at both 20° and 37” ~—‘ Facultative anaerobic.

CLASSIFICATION OF DAIRY BACTERIA. 153

Ps. sapolactica Eichholz, An acid, non-liquefying Pseudomonas (soapy).
This organism was sent by Weigmann and was stated by him to produce soapy-
tasting milk. When studied by us this latter character was not apparent. We
have found an identical organism (not soapy) in New York city milk, and twice
in Middletown. The Middletown culture was found in great quantities in
some milk that had been preserved at 1° for several weeks. A very closely
allied organism was sent us from Geneva by Harding, which we have called
variety A.

Morphology.—Size, .8u-1.74 x .74-.84. No chains, no spores, no capsules,
Gram stain negative.

Gelatine colony.—A round, raised, smooth, homogeneous, entire colony, of.
agray color. In itmus gelatine, in some cases, the colony becomes acid.

Gelatine stab.—A filiform needle growth, and a flat surface.

Agar streak,—Filiform, thick, smooth, opaque, white, moist, moderately
luxuriant.

Fermentation tubes.—Dextrose only is rendered acid. There is no gas or
closed arm growth in any sugar bouillon,

Bouillon.—A sediment, decided turbidity, and a ring-like pellicle.

Milk.—Is rendered alkaline, at least after several days, but there is no other
change.

Potato.—A slight growth, linear, thin, contoured, brownish in color, moist.
Potato not discolored.

Grows rather better at 37° than at 20°. Aerobic.

Variety A,—Is acid in all three sugar bouillons, curdles milk and shows no
potato growth.

Il. Liquefying.

Ps. lactis ananan.s, This was originally described in 1899. Several cul-
tures from milk in Middletown, and New York, and Storrs, have been found
agreeing with this in all points except in the presence of the banana smell upon
potato. The following more complete description was from a culture isolated
more recently from milk in Middletown:

Morphology.—Size, .8u-1.2p x .5¢. Forms chains, no spores and no capsules.
Gram stain negative.

Gelatine colony.—A very rapid liquefier, producing a granular pit; not
characteristic.

Gelatine stab,—Liquefes rapidly; either stratiform or infundibuliform.

Agar streak.—Spreading, flat, smooth, creamish to brown color, rather
luxuriant, and sometimes slightly viscous.

Fermentation tubes.—No acidity nor gas in any sugar bouillon. One culture
showed growth in the closed arm, but the others did not.

Bouillon.—A granular sediment, and a decided flocculent turbidity, but no
pellicle. The liquid is sometimes but not always greenish.

154 STORRS AGRICULTURAL EXPERIMENT STATION.

Milk.—Is curdled without change in the reaction, or slightly alkaline, and is
afterwards digested. After digestion it is colorless and transparent, and some-
times jelly-like. It may have a greenish tinge.

Potato.—A luxuriant growth, widely spreading, convex, smooth and moist, of
a brownish color. Potato not discolored.

Grows at both 20° and 37° _— Aerobic.

Probably the culture that produces the green color deserves to be called a
distinct variety.

Ps. lactis Eurotasn.s. A brown, fluorescent, liguefying Pseudomonas.
This organism has been found in milk directly from the udder and in New
York city milk, the latter differing from our organism in the points indicated by
brackets.

Morphology.—Size, .gu-1.54 x .34. No chains, no spores, no capsules, and
Gram stain negative.

Gelatine colony.—A round, convex, smooth, punctate, entire colony, of a
gray-brown color. On Atmus gelatine it is alkaline and slowly liquefies.

Gelatine stab.—Liquefaction is slow; stratiform.

Agar streak,—A \axuriant growth, linear, flat, smooth, gray [yellow], moist.
The agar shows an amber-colored fluorescence.

Fermentation tubes.—All three bouillons are rendered alkaline, and show
closed arm growth, but no gas.

Bouillon.—A sediment, a decided turbidity, and a flocculent pellicle.

Afilk.—\s rendered alkaline, is curdled both at 20° and 37°, and is completely
digested. [The milk does not curdle.]

Potato.—A luxuriant, linear, smooth growth, of a brown color [yellowish].

Grows at both 20° and 37°. Facultative anaerobic.

Ps. lactis nigra Gorini. A black pigment forming Pseudomonas. This organ-
ism was received from Gorini, and the characters as given below were determined
by him. It clearly resembles 2B. lactis niger A.

Morphology.—Size, 2u-3.54x Iu. No chains, no spores, no capsules. Gram
stain negative.

Gelatine colony.—A pit, liquefying slowly, and with an irregular centre.
Gelatine stab.—Liquefaction begins in twelve hours; infundibuliform.
Ayar streak.—A filiform, raised, rugose growth; opaque, cream-white.

Luxuriant, with a dull surface, and sometimes wrinkled. The agar shows a
brown fluorescence.

Fermentation tubes.—No acidity nor gas in any sugar bouillon. A very slight
growth in the lower quarter of the closed arm.

Bouillon.—A sediment and slight turbidity, with a wrinkled pellicle, which
may sink in flakes to the bottom.

Mitk.—Rendéered acid, curdled, and subsequently digested. The digestion
is complete and a slight sliminess is produced.

CLASSIFICATION OF DAIRY BACTERIA. 155

Potato.—Luxuriant, spreading, convex, smooth, grayish-brown. Potato
chocolate-colored.

Grows well at 20° and 37°. Aerobic.

Ps. lactis contorta n. >. A polypiform, spore-bearing, monotrich, This
organism has been found only once, in milk from New York.

Morphology.—Size, 1-54 x .84. It apparently produces spores and has a
single flagellum, although it was lost before the study was completed.

Gelatine colony.—A slow, liquefying pit, at frst umbonate. On dtmus gela-
tine it is highly lobed and contorted, and does not liquefy in four days.

Gelatine stab—A filiform needle growth, with a flat surface growth, which
later becomes a dry pit. True liquefaction does not appear.

Agar streak,—A moderate growth, filiform, convex, smooth, opaque, of a
cream-white color.

Fermentation tubes.—No acidity nor gas in any sugar bouillon, but growth
in closed arm.

Bouillon.—A sediment, slight turbidity, and a granular pellicle.
7 Milk.—Rendered alkaline, but not curdled. At 20° there is a slight diges-
tion and the milk is slightly slimy.

Potato.—Luxuriant growth, filiform, convex, smooth, opaque, of a gray color.

Grows better at 20° than at 37° Facultative anaerobic.

Ps. lactis minuta n. >. A very small spore-bearing monotrich. This
organism has been found only once, in milk directly from the udder.

Morphology.—Size, .6u-.8u x .3#. A very short rod, not forming chains.
No spores, no capsules. Gram stain positive.

Gelatine colony.—Round, raised, smooth, entire colonies, of a brownish-
yellow color, which, later, liquefy.

Gelatine stab.—A slow liquefaction, crateriform.

Agar streak.—Luxuriant, filiform, raised, translucent, porcelain-white, moist.

Fermentation tubes.—No acidity, gas, nor closed arm growth in any sugar
bouillon.

Bouillon,—A flocculent sediment and a slight turbidity, but no pellicle.

Milk.—Rendered acid, is not curdled or digested, and no other change is
noticeable.

Potato.—No visible growth.

Grows at both 20° and 37° = Aerobic.

‘

Ps. lactis mina n. ». A Gram-staining, spore-bearing, monotrich, This
organism has been found only once, in New York city milk. It is somewhat
similar to the last, but differs in some important points.

Morphology.—Size, 1.4u-1.84 x .6u. A slenderrod. No spores, no capsules.
Cram stain negative.

156 STORRS AGRICULTURAL EXPERIMENT STATION.

Gelatine colony.—A slow liquefying pit, or sometimes a dry pit without lique-
faction, but with a dense colony at the bottom.

Gelatine stab.—No true liquefaction, but a deep, dry pit is formed.

Agar streak. —A luxuriant growth, filiform, raised, smooth, opaque, white,
moist.

Fermentation tubes.—No acidity, gas, nor closed arm growth in sugar
bouillon.

Bouillon.—A granular sediment and decided turbidity, with a membranous
pellicle. 7

Milk.—No change except a slight alkalinity.

Potato.—A nodose, convex, contoured growth, gray in color, moist, and not
luxuriant. The potato is discolored.

Grows better at 20° than at 37°. Aerobic.

Ps, lactis Robertii n. s. A white, non-acid, liquefying :monotrich, This
organism was found in Middletown and New York city milk. A second
variety, found at Storrs, differed in points shown in brackets.

Morphology.—Size, 24x .7u-.94 [1m x 3H]. Large rods with square ends.
No chains. Gram stain positive. No capsules and no spores.

Gelatine colony.—A rapidly liquefying colony, with a greenish-orange pig-
ment. [Brownish.}

Gelatine stab.—A rapid liquefier, stratiform, with a clear yellow liquid. [Slow
liquefier. ]

Agar streak.—Luxuriant, raised, smooth, moist, white. [Dry and wrinkled. ]

Fermentation tubes.—No acidity or gas in any bouillon, but growth occurs in
closed arm in all cases.

Bouillon.—A sediment, decided turbidity, and membranous pellicle.

Milk.—Rendered alkaline, curdled and digested. It has an odor and a green-
ish color. [Greenish color and developed. ]

Potato.—Moderately luxuriant, filiform, flat, smooth, brown. [Spreading
flesh color. ] :

Grows at both 20° and 37°. Facultative anaerobic.

The Storrs type should, perhaps, be called variety A. The culture from New
York did not produce greenish milk. ‘

Ps, lactisaurean.s. A yellow, liquefying monotrich. Found only once, in
New York city milk.

Morphology.—Size, 1.4u.x Iu. No spores, no capsules, no chains. Gram
stain positive.

Gelatine colony.—A slow liquefier, with a dark-ringed colony; round. On
litmus gelatine it is lobed at the edge and not acid.

Gelatine stab.—A filiform needle growth, with a flat surface. Liquefaction
begins in one day; stratiform.

Agar streak.—Luxuriant, filiform, raised, papillate, of a lemon-yellow color.

CLASSIFICATION OF DAIRY BACTERIA. 157

Fermentation tubes,—No acidity, gas, nor closed arm growth in any sugar
bouillon.

Bouillon.—A sediment, slight turbidity, and ring-formed pellicle.
Milk.—Rendered alkaline, but no other change noticed.

Potato.—A luxuriant growth, spreading or beaded, contoured; of a lemon-
yellow color, moist.

Grows better at 20° than at 37° Aerobic.

Ps. lactis aerogenes A.u.s. A gas-producing, liquefying monotrich, This
was found in milk which had been kept at 1° for several weeks. Possibly this
is Ps. coadunta of Wright. y

Morphology.—Size, 1v-1.2u x .7¥-.gu. A short rod, with no chains. No
spores, no capsules, and Gram stain negative.

Gelatine colony.—Round, raised or ‘flat, smooth, homogeneous, with wavy
edge. Cream-white to brownish color. On /tmzs gelatine the colony is large,
moist and acid, like a typical B. aerogenes.

Gelatine stab,—A very slow liquefaction. There is a beaded needle growth,
a moderately thick surface, rather rough, and, after a long time, a liquefaction.

Agar streak,—Thin, whitish, extending over the whole surface of the agar;
not characteristic.

Fermentation tubes.—Acid, gas, and closed arm growth in all sugar bouillons.

Bouillon,—A sediment, slight turbidity and a membranous pellicle.

Milk,—Rendered acid and promptly curdled. A very slight digestion, with
a strong odor. ‘The digestion not always apparent,

Potato.—Usually scanty, thin and of a porcelain-white color.

Grows better at 20° than at 37° Facultative anaerobic.

Ps. fluorescens Gorini, A fluorescent monotrich. This organism, received
from Gorini, had the following characters when studied by us. A similar
organism from Colchester milk differed in points shown in brackets.

Morphology.—Size, 1.2u-1.84x .74%. No chains, no spores, no capsules.
Gram stain negative.

Gelatine colony.—A moderately transparent, liquefying colony, granular and
cloudy. It liquefies the whole plate in time. Litmus gelatine not acid.

Gelatine stab.—It begins to liquefy in one day, infundibuliform, with a
tenacious scum. ([Stratiform.]

Agar streak,—Luxuriant spreading, raised, smooth, opaque, gray. [Thin and
flat.] The agar shows a green fluorescence.

Fermentation tubes.—Dextrose bouillon is acid, but the others are not. No
gas in closed arm growth in any case.

Bouillon.—A sediment and a decided turbidity, but no pellicle. [Pellicle is
formed. ]

Milk.—Becomes alkaline at 20°, is curdled and completely digested, but not
at 37° Shows a green color and strong odor. [Curdles and digests at 37°,
without the green color.]

158 STORRS AGRICULTURAL EXPERIMENT STATION.

Potato.—A luxuriant, spreading, thin, white growth. Potato discolored.
{Thick.]
Grows better at 20° than at 37° Facultative anaerobic.

Ps. lactis varians n.s. An acid-producing, white monotrich, This organ-
ism, incompletely described in 1899, is now more completely described from
recently isolated cultures that are probably the same. ‘They are quite common

_and have been found in abundance in milk that had been preserved for several
weeks at I°.

Morphology.—A rod, forming chains; the individual elements Im-1.4u x .8p.
No spores, no capsules. Gram stain negative.

Gelatine colony. —A round, flat or umbilicate, smooth, rugose colony, of a
slight brown, yellow or cream-brown color, producing a slow liquefaction.

Gelatine stab,—Stratiform or infundibuliform. The liquefaction % slow, and
one culture produced a dry pit without liquefaction.

Agar Streak.—Filiform, raised, smooth, opaque, white and moderately
luxuriant.

Fermentation tubes.—Usually acid in dextrose but not in other sugar bouillons.
No gas nor closed arm growth in any bouillon.

Boutllon.—A sediment, an abundant turbidity, and a membranous pellicle.

Milk,—Rendered slightly acid and curdled at 20°, but not at 37° No
visible digestion, but a prominent odor.

Potato.—Somewhat variable. It may be thin, spreading, smooth and gray-
brown, or more luxuriant, varying from white to brown.

Grows better at 20° than at 37°, though there is a slight growth at 37°.
Aerobic. 7

Variety A.—One culture of this organism differs from the above description
in the following points: Size, 1¢x.5. Liquefies rapidly, beginning in one
day. The colony forms a granular liquefying pit. Saccharose bouillon acid
and shows growth inclosed arm. Milk rendered alkaline, curdled, and digested.
A luxuriant, thick growth on potato.

Here belong, probably, Gorini’s 8. actdificans presamigenes casei and
Griiber’s Ps. fragagariae //, (Cent. f. Bact. II., XIV., 122), neither of which
is thoroughly described. Griiber’s organism is not sufficiently described for
identification. Gorini’s has the following characters:

B, acidificans presamegemus casei Gorini. (Rev. gen. d’lait, III., 505.)

Morphology.—Size, 8u-top. x 24 (?) Produces spores and stains by the Gram
method. Though not distinctly stated, this is probably monotrichic.

Gelatine colony.—A round, white, filamentous, irregular colony, which lique-
fies gelatine.

Agar streak.—A luxuriant, white growth,

Lrermentation tubes.—Dextrose bouillon is rendered acid and shows closed
arm growth.

Bouillon,—A sediment, turbidity, and pellicle are produced.

ty

CLASSIFICATION OF DAIRY BACTERIA. 159

Milk,—Rendered acid, curdled, and digested at 20°, without change in color.
Potato.—Grows well.

Grows at 20° and 37°.

Ps, lactis granulan. s. A non-curdling, liquefying monotrich. Found in
New York city milk and also sent by Harding.

Morphology.—Size, .74x 24. A rod, forming chains. There are no capsules,
and the Gram stain is negative. Spores are produced.

Gelatine colmy.—A rapidly liquefying pit is formed, which is uniformly
coarsely granular, and has a ciliated margin.

Gelatine stab.—A spiny needle growth and a napiform pit, which later is
stratiform. Liquefaction begins in one day.

Agar Streak.—A filiform, pie smooth, grayish, moderately luxuriant
growth.

Fermentation tubes.—Acidity is produced in all three bouillons, but no gas
nor closed arm growth.

Bouillon.—A sediment, an abundant turbidity, and a membranous pellicle.

Milk.—Rendered slightly alkaline, but no other change seen.

Potato.—Shows no growth.

Grows hardly at all at 20°, but abundantly at 37°. Aerobic.

THE GENUS BACILLUS, LOPOTRICHIC.

B. syneyanus (Ehrb.) Migula=cyanogenes Fligge. Bacillus of blue milk.
We have not found this species, but a culture was sent to us by Duckwall. Its
characters, as determined by us, are as follows:

Morphology.—Size. I.2u-2u x .54. Short chains; spores produced, but no
capsules. Gram stain irregular. It possesses a tuft of flagella at one end.

Gelatine colony.—A round, raised, smooth, entire colony, of a grayish color,
-5 mm. in diameter, in three days.

Gelatine stab.—A filiform, needle growth, and thin surface, which does not
spread. After a few days the gelatine turns a dark color at the surface, but
does not liquefy.

Agar streak.—Luxuriant, spreading, thin, smooth growth, translucent, white,
and showing a dark, smoky fluorescence.

Fermentation tybes,—Dextrose and saccharose, rendered alkaline, without
gas or closed arm growth, and no change in color. Lactose becomes very
slightly acid, without gas or closed arm growth, and turns a deep blue-black.

Boutllon.—A black sediment, and dark-colored turbidity, with a membranous
pellicle.

Milk.—Rendered slightly alkaline without curdling, but develops an odor, and
after a few days becomes distinctly blue.

Potato.—A very luxuriant, spreading, thick growth, translucent or opaque,
brownish in color. ‘

Grows at both 20° and 37°. Aerobic.

160 STORRS AGRICULTURAL EXPERIMENT STATION.

B. lactis olivaceus n. s, A greenish Peretrich, This organism has been
found in milk directly from the udder.

Morphology.—Size, 1.5472" x .4h. A small rod. No chains, no spores, no
capsules. Gram stain negative. It has a tuft of flagella at one end.

Gelatine colony.—A round, convex, smooth, homogeneous, entire colony, of
a reddish color below the surface. The surface colony has an irregular outline,
and a reddish or a greenish color.

Gelatine stab.—No liquefaction, but a needle and surface growth.

Agar streak,—Luxuriant, filiform, raised; smooth, greenish in’ color.

Fermentation tubes.—No acidity, gas, nor closed arm growth in any sugar
bouillon.

Bouillon,—A sediment, an abundant turbidity, and a granular pellicle formed.

Milk.—Rendered alkaline, becomes greenish, and develops a strong odor
without curdling. .

Potato.—A luxuriant, filiform, flat, smooth growth, brownish-yellow.

Grows both at 20° and 37° —‘ Aerobic.

B. lactis minutus n.s. A yellow Lophotrich. This organism was isolated
from milk directly from the udder.

Morphology.—Size, .5@x .4u. An extremely minute rod, which forms short
chains. It produces no spores, no capsules, and does not accept the Gram
stain. _ Several flagella at one end.

Gelatine colony.—A round, convex, smooth, homogeneous colony, of a red
color. On Aitmus gelatine it is not acid.

Gelatine stab.—A needle growth, but no surface growth.

Agar streak.—Filiform, raised, smooth, translucent, yellow, moist, luxuriant.

Fermentation tubes.—No acidity, nor closed arm growth in any sugar
bouillon.

Bouillon,—A sediment and turbidity, but no pellicle.

Milk.—No action.

Potato,—Very scanty growth, yellowish.

Grows at both 20° and 37°. Aerobic.

B. lactis molocularis n. >. A white, non-liguefying Lophotrich. Found in
New York city milk.

Morphology.—Size, 1.44 x .7#. A lophotrichic rod with flagella at both ends,
No spores, no capsules. Gram stain negative.

Gelatine colony.—An opaque bead, smooth, entire, white. On Atmus gelatine
an extremely diffused growth, made up of microscopic dots, appearing to the
naked eye simply as a cloud extending over the plate. The individual colonies
are visible only under the microscope.

Gelatine stab.—A filiform needle growth and a flat surface, without liquefac-

tion.

CLASSIFICATION OF DAIRY BACTERIA. I61

Agar streak.—A filiform, flat, smooth growth, of a gray-white color, rather
scanty. ,

Fermentation tubes.—Dextrose is rendered acid, but there is no acidity on
other sugar bouillons, and there is never any gas or closed arm growth.

Bouillon.—A sediment, turbidity and a pellicle.

Milk,— Rendered alkaline, and is curdled at 37°. There is no digestion, but
an unpleasant odor,

Potato.—A scanty, flat growth, moist, smooth, of a brownish color.

Grows better at 20° than at 37° Aerobic.

B. lactis Isigniin.s. A brown, fluorescent Lophotrich, Found in Isigny
cheese, comprising 18% of the bacteria in the centre of the cheese.

Morphology.—Size, .64 x .ou. A rod, occasionally forming chains. Spores
are developed, capsules are present, and Gram stain is positive. Usually one
flagellum is present, though sometimes a tuft is seen at one end.

Gelatine colony.—An extremely rapid liquefier, without any distinct character-
istic.

Gelatine stab.—Begins to liquefy in two days, and is complete in twelve days.
Sacchate or stratiform.

Agar streak.—Spreading, thin, smooth, luxuriant, translucent white. At
37° the agar becomes brownish, and at 20° it becomes at first flesh color and,
later, brown. :

Fermentation tubes.—Acidity, gas and closed arm growth in all sugar
bouillons.

Bouillon.—A sediment and decided turbidity, but no pellicle.

Milk.—Rendered acid and curdled at both 20° and 37°, but no subsequent
digestion noticeable.

Potato.—A luxuriant, thick, smooth growth, of a yellowish color, and a dis-
colored potato. .

Grows at both 20° and 37°. Aerobic.

Il. Liquefying Lophotrichic Bacilli.

B, lactis fluorescens In. s. A lophotrich producing a smoky fluorescence and
a green color,

Morphology.—Size, 8u-1.6ux .4u-.6p. A lophotrichic rod with two or three
polar flagella. Nochains. Spores developed in the ends of the rods. Gram
stain negative, and no capsules.

Gelatine colony.—A round, gray, smooth, white colony, which liquefies into
a uniformly granular pit.

Gelatine stab.—Liquefies rapidly, infundibuliform, with a cloudy liquid, full
of flakes.

Agar streak, —Luxuriant, filiform, smooth, raised, gray, moist. The agar is
smoky.

162 STORRS AGRICULTURAL EXPERIMENT STATION.

Fermentation tubes.—Dextrose bouillon is rendered acid, but there is no gas
nor closed arm growth, nor is there acidity in any other sugar bouillon, In all
cases the liquid is turned green.

Bouillon.—A granular sediment and turbidity, but no pellicle. The bouillon
is green after a few days.

Milk.—No change in the reaction, but the milk is curdled and digested at
both 20° and 37°, and becomes green in color, with a strong odor.

Potato.—Scanty, filiform, flat, smooth, brownish-yellow.

Grows at both 20° and 37°. Aerobic.

B. lactis fluorescens II. n. s. A green fluorescent lophotrich. This is a
common form. The culture from which the following description was made
was isolated from Camembert cheese. Some variations are shown in the points
in brackets.

Morphology.—A \ophotrichic rod, sometimes forming chains. Individual
elements, Iu-1.44x .64-.94. No spores nor capsules. Gram stain negative.

Gelatine colony,—A very rapid liquefier, forming a cloudy pit. [A bead anda
granular pit.] On Atmus gelatine it is strongly alkaline.

_ Gelatine stab.— Begins to liquefy in about three days, napiform. [Infundibuli-
form.] The liquid shows fluorescent green, and there is a surface membrane.

Agar streak.—Luxuriant, filiform or spreading, raised, smooth, of a white or
brownish color. The agar turns green. The depth of the green color varies
in different cultures. [Growth on agar in one culture is green, as well as the
agar itself. ]

Fermentation tubes.—Dextrose bouillon is acid, but no other sugar bouillon,
and there is no gas, nor closed arm growth. [Dextrose not acid.]

Bouillon.—A sediment, a decided turbidity, and a flocculent pellicle. The
liquid is green.

Milk.—Is alkaline and curdled at 20° and subsequently digested, green. Has
a pleasant odor.

Potato,—Scanty, flat, smooth, brownish. [Luxuriant.]

Grows at both 20° and 37°, but better at 20°. Aerobic, or facultative
anaerobic.

B. fluorescens minutissinus. A fluorescent lophotrich. This organism,
described in 1899, is probably identical with the last. It shows slight variations
and its description is reinserted here.

Morphology.—An extremely small rod. Size, .5u-.74x.54. No chains, no
spores, no capsules. Flagella were not made out.

Gelatine colony.—A smooth, liquefying pit, without a nucleus, but granular.
Gelatine stab.—Liquefies stratiform, with a cloudy liquid, which is not green.
Agar streak.—Luxuriant, soft white growth, with a green fluorescence.

Bouilion.—A sediment and turbidity:are formed, with a pellicle. In two days
the bouillon is very cloudy, but not green.

r
t
‘
I
}

«

CLASSIFICATION OF DAIRY BACTERIA. 163

Milk.—The milk is curdled with a slight green color at the top, but there is
no apparent digestion.

Potato,—A luxuriant, white to brownish growth, with no discoloration.

Grows at both 20° and 37°, but the green color does not appear at 37”.
Aerobic.

B. lactis fluorescens III. n. s. A green, non-fluorescent lophotrich. This
organism does not produce a fluorescence, but the fact that it turns milk green
suggests the relationship to the last organisms, and hence we class it here.

Morphology.—A slender rod.* Size, 1.54-34x .4u-.74. No chains, no
spores, no capsules. Gram stain negative.

Gelatine colony.—A very rapidly liquefying colony, uniformly granular, but
not characteristic.

Gelatine stab,— Begins to liquefy in two days, stratiform.

Agar streak.—Luxuriant, spreading, flat, smooth growth, gray-brown, moist.

Fermentation tubes.—Acidity and closed arm growth in dextrose and some-
times lactose, but not in saccharose. No gas produced. The liquid is usually,
though not always, green.

Bouillon.—A sediment, slight turbidity, and a membranous pellicle.

Milk.—Becomes alkaline, curdles, and digests. The digested liquid is green
or yellow in color, and has a strong odor.

Potato.—Growth scanty, but the potato is discolored.

Grows at both 20° and 37°. Facultative, anaerobic.

B. lactis moruloideus n. s.

Morphology.—Size, 1u-1.54x 1.24. No chains, spores, or capsules. Gram
stain irregular, frequently showing only a single flagellum.

Gelatine colony.—A slowly liquefying pit, which is lobed and moruloid, with
a putrefactive odor. In “tmus gelatine it is not acid.

Gelatine stab.—A needle growth, and a stratiform liquefaction, beginning in
one day, and never complete.

Agar streak,—Filiform, raised, smooth, rather opaque, white, moist, not
luxuriant.

Fermentation tubes.—Dextrose is acid but no other sugar bouillon. Occasion-
ally growth in closed arm, but no gas.

Bouillon.—A sediment, turbidity, and a pellicle.

Milk.—Is rendered acid and curdled with a subsequent digestion, with a
strong odor and a yellowish color.

Potato.—Scanty, thin, smooth, white, moist; no discoloration.

Grows at 20°; very slightly at 37° Facultative anaerobic.

* Frequently appearing monotrichic, from the breaking away of some of the flagella.

164 STORRS AGRICULTURAL EXPERIMENT STATION.

THE GENUS BACILLUS, PERITRICHIC, NON-LIQUEFYING,
I. No acid in dextrose or other sugars.

B. lactis nigroferous n. s. A black bacillus. This bacillus was obtained
from milk in New York city, and is distinctly characterized by its /ue-black,
nearly jet-black color. B. niger of Biel, and a species isolated by Gorini, pro-
duces black colonies on gelatine, but not on agar. We have seen only one
culture of this remarkable bacillus, which is, so far as we know, the only bac-
terium described of a jet-black color. :

Morphology.—A peritrichic rod. Size, .qu-1ux.gu. There are no spores,
and the organism does not accept the Gram stain. No chains are formed.

Gelatine colony.—A round, thin, smooth, moist colony, at first white in color,
but later a very deep blue-black. On Atmus gelatine, black; in a pit, not acid.

Gelatine stab,—A needle growth and a good surface growth, slightly arbores-
cent at the top, becoming black.

Agar streak.—A moderately thick, smooth, moist growth, which becomes
jet-black.

Fermentation tubes.—There is growth in the closed arm in all three sugar
bouillons, but no acid is produced, and no gas. An indigo blue scum appears.

Bouillon.—An abundant sediment, a turbidity, and a blue-black pellicle.

Milk.—No change, except the formation of a thick, black scum. Later the
milk becomes black.

Potato.—A thick, spreading, moist, very luxuriant growth, of a deep blue-
black color; potato discolored.

Grows at 37° and 20°, but not well at 37°. Aerobic.

We have several times isolated from milk bacilli that seem essentially identical
with B. Zenkeri (Hauser). They have been found in milk from several localities
in Connecticut. This essential agreement is indicated by the name we have
given it. The culture as studied by us has the following characters:

B. lactis Zenkert n, s. (Hauser). A rhizoid or proteus-like bacillus.

Morphology.—Size, 24-34x 14. Frequently in chains. It produces no
spores, and does not accept the Gram stain.

Gelatine colony.—A peculiar rhizoid colony is formed, with lateral extensions
of variable character.

Gelatine stab.—A prominent needle growth and a lobate or polypiform surface.

Agar streak,—A thick, luxuriant, white growth, with radiating fibres from a
ragged edge. Its surface is dull. Sometimes viscous. ‘

Fermentation tubes.—No acidity, gas, nor growth in closed arm in any sugar
bouillon.

Bouillon.—A sediment, but no pellicle, and usually no turbidity.

Mitk.—Unchanged, except for a slight alkalinity. Some specimens, however,
show a slight sliminess.

Potato.—A moderately thick, dirty white or brown growth, which is apt to be
rough and dry, Sometimes it is yellow, and the potato is discolored.

CLASSIFICATION OF DAIRY BACTERIA. » 165

Grows both at 20° and at 37°, but better at 37°. Aerobic.

One culture of this organism which we have found, spread over the surface of
a gelatine stab as a thick felt, but on potato it produced no growth, The
characteristic rhizoid colony, however, leads us to place it here.

B. lactis Colchesterii n. s. A yellow, rhizoid, peritrichic bacillus.

Morphology.—Size, I~-.2ux.74-.94, forming short chains. It accepts the
Gram stain, and has an evident capsule, but produces no spores.

Gelatine colony.—A thizoid colony is produced that looks exactly like a mold.

Gelatine stab,—There is a needle growth and a surface growth.

Agar streak.—Mold-like colonies are formed which extend under the surface
of the agar. The growth is luxuriant and of a yellow color. Frequently with
an iridescence.

Fermentation tubes.—No acidity, gas, nor growth in closed arm in any sugar
bouillon.

Bouillon.—An abundant sediment, a slight turbidity, but no pellicle.

Mitk.—No action on milk.

Potato.—A thin, yellow growth, not widely spreading.

Grows at both 20° and 37°. Aerobic.

Only a single culture of this organism has been found in milk, from Col-

chester, Conn. It is readily distinguished from other bacilli by its mold-like
colony, and its yellow color.

B. lactis nebulusn.s. A smoky bacillus.

Morphology.—A very small rod. Size, .8u x .3#. It forms chains, is actively
motile, produces no spores, and does not accept Gram stain.

Gelatine colony.—A thick, contoured, smooth colony of a yellow color. On
litmus gelatine a plain white colony is formed, not characteristic.

Gelatine stab.—An abundant needle growth and a transparent surface
growth.

Agar streak.—A luxuriant, rather thick, smooth growth, opaque, white. The
agar shows a smoky fluorescence.

Fermentation tubes.—Neither acidity, gas, nor growth in closed arm in any
sugar bouillon.

Bouillon.—An abundant, amorphous sediment, and a slight turbidity, with a
pellicle on the surface.

Milk.—No action.

Potato.—A thin, scanty, white growth.

Grows better at 20° than at 37°. Aerobic. "

Two cultures have been found which we have carefully studied and regarded
as probably the same. One of them was isolated in 1895 and the other in 1903°
The former did not produce the smoky fluorescence in agar, nor did it produce
a pellicle on bouillon. In other respects they were the same.

166 STORRS AGRICULTURAL EXPERIMENT STATION.

Il. Acid in Dextrose or other Sugars.

B. lactis citreusn, s. A yellow, non-liguefying perttrich,

Morphology.—Our observations were incomplete when the culture was lost.
It is a peritrichic (?) rod, .84x .5@, which forms no chains or spores.

Gelatine colony.—A white, opaque colony, that later becomes yellow, 2 mm.
in diameter.

Gelatine stab.—A needle growth and a lemon-yellow surface growth; um-
bilicate.

Agar streak.—A laxuriant, lemon-yellow growth, smooth, moist.

Fermentation tubes.—Probably acid without gas.

Bouillon.—A sediment, a turbidity, and a pellicle, the latter sinking to the
bottom.

Milk.—Becomes acid and curdles into a hard curd, with a layer of liquid on
top.

Potato.—A luxuriant growth which is at first white and then lemon-yellow.

Grows at 20° and 37”. Aerobic.

B. lacto rubifaciens. Gruber. A red pigment bacillus. This culture was
sent us from Kiel by Weigmann. The culture when received and studied in our
laboratory, produced no color except a slight pinkish tint in milk.

Morphology.—An active rod, 24-34.x.74. No chains are formed; it does not
accept Gram stain. Spores are formed, but no capsule.

Gelatine colony.—Thick, contoured, gyrose, white. On “tmus gelatine it be-
comes 5 mm. in diameter, is acid and lobed, with gyrose and with a mottled
surface.

Gelatine stab.—A good needle growth, villous, with a spreading surface.
Agar streak.—Linear, moderately thick, white.

Fermentation tubes.—All three sugars develop acidity, and show growth in
the closed arm, but no gas is produced.

Bouillon.—A flocculent sediment, a turbidity, and a ring-formed pellicle.

Mitk.—Rendered acid and curdled into a gelatinous mass at 20°, but not at
37°. When heated the jelly becomes a hard curd, with a jelly-like whey. The

milk has an odor of the barn, and a pinkish color.
Potato.—Rather thick, white, luxuriant. Potato discolored.
Grows at both 20° and 37°, though better at 20°. Facultative anaerobic.

B. lactis sulcatusn. >». A non-gas-producing, acid bacillus, without spores.
This organism has been found twice at intervals of three years. In one
case it came from market milk, and in the other directly from the udder. There”
were slight differences in the colonies of the two cultures, and one culture failed
to curdle milk even when heated. They seem to agree fairly well with a cul-
ture from cream descyibed by Severin (Cent. f. bact. II., XI., 1903, p. 202). We

name it from its resemblance to B. sulcatus.

CLASSIFICATION OF DAIRY BACTERIA. 167

Morphology.—An active rod. Size, 24-2.5ux.6m¢. No chains nor spores,
It accepts the Gram stain.

Gelatine colony.—A large [6-8 mm.], spreading, white colony, with a rough,
irregular, contoured surface, outline indented. On “itmus gelatine the colony
is acid.

Gelatine stab.—A needle growth and a very thin surface growth.

Agar streak.—Thin, linear, white, rather scanty, and not characteristic.

Fermentation tubes.—All three sugar bouillons show acidity and growth in
the closed arm, but no gas.

Bouillon.—A flocculent sediment, but no turbidity nor surface pellicle.

Milk.—Becomes acid but does not curdle unless heated.* No other change.

Potato.—Thin, scanty, moist, white.

Grows both at 20° and 37° ‘Facultative anaerobic.

B. aromaticus lactis Grimm, (Cent. f. Bac. u. Par, II., VIII., 584, tg02) seems
to belong here.

For peritrichic gas-producing acid bacilli see p. 182.

B. disenteriae Shiga. This organism, regarded as the cause of some forms
of dysentery and some cases of summer complaint, we insert here, although, so
far as we know, it has not actually been found in milk. It is strongly suspected,
however, that it is sometimes distributed by milk, and we have thought it well
to include it in our list. The characters as given below are described by Ved-
der and Duval.

Morphology.—A peritrichic rod, 1u-3u in length, and very slender. Some-
times it is extremely short, almost a coccus. It produces no chains nor spores.
It does not accept the Gram stain.

Gelatine colony,—Practically identical with the colony of B. coli communis.
A thin, slightly spreading, white colony, which is acid in litmus agar.

Gelatine stab.—There is a needle growth and a slight surface growth which
does not spread.

Agar streak.—A luxuriant, uneven, rather thick, cream-white growth, which
later shows a feathery edge.

Fermentation tubes.—All sugar bouillons are rendered acid but no gas is pro-
duced.

Bouillon.—A sediment and turbidity are formed, and occasionally a thin
pellicle; later the liquid is clear.

Milk. — At first acid, but later alkaline. No curdling nor other change.

Potato.—A luxuriant, rough, thick, spreading, yellowish growth.

Grows better at 37° than at 20°. Produces indol, and is pathogenic.

This is very similar to B. colz, but grows less rapidly at 37° They are not
easy to separate from each other, special culture methods being necessary

B. lactis fragariae (Weig.). This culture was sent me by Weigmann,
labeled Pseudomonas fragariae, The culture we have received is a peritrichic
bacillus rather than a Pseudomonas, and does not produce any peculiar odor in

168 STORRS AGRICULTURAL EXPERIMENT STATION.

milk. Whether this is due to a contamination and thus to a loss of the original
culture, or whether it has changed its characters, there was no way of determin-
ing. The culture which we received shows the following characters:

Morphology.—A bacillus 1.34-1.54x .74-.94. It forms no chains nor spores,
and does not accept the Gram stain.

Gelatine colony.—A round, thick, smooth, homogeneous, entire colony of a
white color. On tmus gelatine it is nearly transparent, mottled, .5 mm. in
diameter, and gives an odor of ammonia.

Gelatine stab.— A needle growth, and thin surface growth.
Agar streak.—A scanty, thin, smooth, moist, white, growth.

Fermentation tubss.--Acidity is produced in dextrose, but no gas and no
closed arm growth. No effect upon other sugar bouillons.

Bouillon.—A flocculent sediment, decided turbidity, and a pellicle.

Milk.—Is rendered alkaline and slightly transparent, but no other change.
A slight odor is produced.

Potato.—A scanty, thin, smooth, moist, white growth.

Grows at 20° and at 37°. Aerobic.

THE GENUS BACILLUS, PERITRICHIC, LIQUEFYING.
I, Producing Pigment.

B. prodigiosus (Ehrb) Fligge. This well-known organism we have found
many times in milk. So far as we have seen, it never produces any trouble in
the dairy. Its characters are well known, but we insert them here for complete-
ness’ sake.

Morphology.—Size, 5é-1" x .5¢, with chains and coccoid forms. No spores.

Gelatine colony.—Round, oval, entire, reddish-brown, with translucent bor-
ders. Surface irregular and liquefying with production of a red pigment.

Gelatine stab.—Saccate liquefaction, with reddish sediment.

Agar streak.,—White, becoming red.

fermentation tubes.—Glucose is not acid, gas production variable.

Bouillon.—Turbid, a reddish sediment, and a pellicle.

Milk.—Acid coagulated and subsequently digested, with more or less of a
pink color.

Potato.—Rose-red, moist, becoming dark-red to purple.

Grows best at 20°-25°. Aerobic.

B. butyri rubri, Stadling and Poda. (Milch. Zent. II., p. 97, 1906,) A
red, liguefying bacillus, This bacillus, recently isolated from red butter by
Poda, has been carefully studied by him and is quite similar to B. prodigiosus.
It differs, however, very decidedly in its gelatine colony and in its power of
producing red pigment, which is much less than in B. prodigiosus. It produces
red spots in butter, Poda has given it the above name and regarded it asa
distinct type with the following characters:

CLASSIFICATION OF DAIRY BACTERIA. 169

Morphology.—A peritrichic rod. Size, tu-1.5u x .7u-.84. It forms chains
but no spores nor capsules, and does not accept Gram stain.

Gelatine colony.—Round to oval, brown or yellow colonies, with a central
colony in a liquid pit. The colony is opaque and granular, 4 mm. in diameter
in 48 hours, and liquefies at the edge; with a cheesy smell.

Gelatine stab.—A needle growth and a shallow liquefying pit, with no color.
It then becomes infundibuliform, but no red pigment develops.

Agar streak.—A luxuriant, opaque growth, with a central wine-red streak,
and a colorless peripheral zone. Condensation water, dark colored.

Fermentation tubes.—Not given, but probably produces acid and gas.

Bouillon.—A sediment and turbidity. There is no pellicle, but there is a
rose-red color near the surface.

Milk.—Made acid, and curdled with a cheesy smell and a rose-red color. It
is subsequently digested into a yellow liquid with a cheesy smell.

Potato.—A luxuriant, carmine red growth, the pigment appearing at 37° as
well as at 20°.

Grows at 20° and 37°, but the color is not so well developed at the higher
temperature.

B. lactis citronus n.s. A lemon-yellow, peritrichic bacillus. This we have
‘ found but once, in milk fresh from the udder.

Morphology.—Peritrichic bacillus. Size, 1.54x.84. It forms no spores,
accepts the Gram stain and has evident capsules. No chains are formed.

Gelatine ‘colony.—A round, convex, smooth, homogeneous, entire, white
colony, which at liquefaction shows radiation.

Gelatine stab,—Liquefies, infundibuliform.
Agar streak,—Filiform, flat, smooth colony, lemon-yellow color, luxuriant.

Fermentation tubes.—Lactose is rendered acid, but the other sugars are not
acid. Growth in the closed arm shows in all cases, and no gas is produced.

Bouillon.—A sediment and an abundant turbidity, but no pellicle.

Milk.—Becomes acid and is curdled at both 20° and 37° The milk is sub-
sequently digested with an odor, but no color.

Potato.—Spreads over the potato, thin, lemon-yellow, luxuriant.

Grows both at 20° and 37°. Facultative anaerobic.

B. lactis Harrisonii n. s. A slomy milk, yellow bacillus. Isolated by Har-
rison and described by him. (Rev. gen. d’Lait, 1906). I have ventured to
name it after him.

Morphology.—Somewhat irregular. Size, .25¢-.75¢ x .34-3u. No chains
nor spores, and no capsules. It stains by the Gram method.

Gelatine colony,—Irregular, lobulate, slimy, becoming umbonate. Sinks in a
pit 3-7 mm. in diameter, from which the whole colony can be removed by a
needle,

170 STORRS AGRICULTURAL EXPERIMENT STATION.

Gelatine stab.—Outgrowths from the needle track. In two weeks it sinks
into a pit 4 mm. deep, and there is no further change.

Agar streak,—Luxuriant, viscous and dull, at first shiny, later dry; citron
yellow; spreading.

Fermentation tubes.—Not described, but apparently no acid and no gas.

Bouillon,—A turbidity, sediment, and*a ring pellicle.

Milk.—Rendered alkaline, but neither curdled nor digested. It is turned
yellow and becomes slimy.

Potato.—A luxuriant growth, spreading and intensely yellow.

Grows at 20° and at 37°. Aerobic.

B. lactis fluorescens IV. n.s. A fluorescent, liquefying bacillus.

Morphology.—Size, 2.5¢-3.34 X .gu-1.5m@. Forming chains. It produces
central spores, accepts the Gram stain, and shows an evident capsule.

Gelatine colony.—A granular colony with a central nucleus, liquefying almost
immediately.

Gelatine stab.—Liquefying rapidly, infundibuliform.

Agar streak.—Filiform, flat, contoured, opaque, yellowish, moist, and later,
wrinkled. The agar shows a yellow-green fluorescence.

Fermentation tubes.—All three tubes show growth in closed arm; dextrose
alone being acid. No gas produced.

Bouillon.—A flocculent sediment, turbidity, and pellicle. The liquid is
yellow at the top.

Milk.—Rendered alkaline, is curdled at 37°, and is subsequently completely
digested, both at 20° and 37° _—It becomes yellow to orange in color.

Potato.—Spreads completely over the potato, rather thin and yellow, luxuriant.

Grows both at 20° and at 37” _Is facultative aerobic.

Variety A.—Found in milk in Colchester. Differs from the last in the fol-
lowing points. Size, 1.54x.6. Gram stain is negative. The flagella are
extremely long and numerous, and protrude from very thick capsules. Gelatine
stab shows a greenish growth, which later becomes yellow. Yellow pigment is
not produced on agar or on potato. There are no wrinkles on agar and no
growth in the closed arm of fermentation tubes.

The differences between these two varieties are considerable, and are perhaps
sufficient to require recognition under separate names. At present, however,

we leave them together.
«

B. lactis niger (Gorini) n. ». A black liguefying bacillus, This organism,
which was sent us from Geneva by Harding, is almost identical with Ps. lactis
niger of Gorini. See p. 154. The latter, however, is monotrichic, while this
is peritrichic. The other differences are very slight.

Morphology.—Size, 2u-3.54x.g4. Long chains are produced. The organ-
ism.stains with the Gram method, produces no spores and shows no capsules.

CLASSIFICATION OF DAIRY BACTERIA. 171

Gelatine colony.—Not characteristic. A slow liquefier, forming a pit which is
at first clear and then cloudy. No acid is produced upon litmus gelatine.

Gelatine stab.—Begins to liquefy on the first day, infundibuliform. In ten
days the gelatine is about 3 liquefied.

Agar streak.—Spreading, thin, smooth, opaque, white, moist, and later
wrinkled. Quite luxuriant.

Fermentation tubes.—There is a very slight acidity in all three sugars. No
gas and no closed arm growth.

Bouillon.—After three days there is a sediment, a turbidity, and a membra-
nous pellicle.

Milk.—Is rendered alkaline, curdled, and subsequently digested both at 20°
and at 37°

Potato,—Spreading, flat, irregular surface or wrinkled, and becoming blue-
black. F

Grows better at 37°, very little at 20° Aerobic.

B. lactis arborescens If, An arborescent spore-producing bacillus. ‘This
organism was found originally in 1899 and a second culture was isolated from
dust in 1904. The original culture was kept in the laboratory for four years”
and then tested again. It was found to agree with the original description ex-
cept that it no longer produced the arborescent growth in gelatine.

Morphology.—Size, 1.54-4ux .84. No chains nor capsules; spores, sometimes
in the ends of the rods and sometimes in the center; Gram stain positive.

Gelatine colony.—Filamentous, I cm, in diameter, of radiating knotted fibers
and sometimes showing secondary radiations from the knots; slowly liquefying.
In Atmus gelatine deep dry pits with radiating filaments.

Gelatine stab.—A dry pit, which later liquefies. Needle growth arborescent.
Agar streak.—Very thin, scarcely visible, covering the whole surface, white.

Fermentation tubes.—No acidity, gas, or closed arm growth in any sugar
bouillon. ;

Bouillon.—A flocculent sediment, a slight turbidity and a tough scum.
Milk.—No action.
Potato.—Thin, not luxuriant, diffuse gray or brown.

.Grows at 20° and 37°. Aerobic.

B. lactis rhizoides n. s. A rhizoid, non-arborescent bacillus.
Morphology.—Size, 34x .84. No chains, no spores, no capsules, Gram stain
negative.

Gelatine colony.—A myceloid colony, siowly liquefying. On “tmus gelatine
the colony is proteus like.

Gelatine stab.—A needle growth and a saccate liquefaction, beginning in three
or four days.

Agar streak.—Spreading, smooth, thin, transparent, white, luxuriant, moist.

Fermentation tubes.—No acidity, gas, nor closed arm growth in any sugar
bouillon.

172 STORRS AGRICULTURAL EXPERIMENT STATION.

Bouillon.—A slight sediment and turbidity, but no pellicle.
Milk.—No action.

Potato.—A very scanty white growth, with discolored potato.
Grows at 20° and very slightly at 37°. Facultative anaerobic.

B. lactis mycoides n. s. Rhizoid, spore-bearing bacilli. This type of bacillus
we have found very frequently. There are some variations in the characters of
the various cultures. The one described below, which we take as a type, was
from Dr. Maher, and has been used by him in numerous inoculation experi-
ments against certain diseases. The variations which we have found in other
cultures are indicated in brackets.

Morphology.—Size, 1u-4u x .6u-1.2u. Long chains produced, spores present,
no capsule, and Gram stain positive.

Gelatine.colony.—A small burr like, rhizoid colony, soon liquefying and form-
ing a pit with a nucleus. [The rhizoid character is not always found, and the
liquefaction may be slow. The color is sometimes yellowish; it sometimes
shows tangled threads like anthrax. ]

Gelatine stah.—An arborescent needle and a cratiform liquefaction. [Infun-
dibuliform without arborescence. ]

Agar streak,—Luxuriant, dull, wrinkled, white, tough. [Yellowish.]

Fermentation tubes.—No acidity, gas, nor closed arm growth in any sugar
bouillon. ;

Bouillon.—A sediment, turbidity, and a pellicle.

Milk.—Rendered alkaline and curdled after a few days. Digested into an
amber colored or yellowish liquid.

Potato,—Luxuriant, velvety, dry, wrinkled, white.

Grows at 20° and 37°. Aerobic or facultative anaerobic. [A ground-glass
like appearance, with liquid under the folds. Sometimes of a pasty consistency. ]

B. subtilis. Is extremely common in milk, though never in great numbers.
While it will grow in sterilized milk, it does not usually thrive in milk contain-
ing lactic bacteria. In old milk it is, therefore, usually overgrown by the lactic
organisms. We describe a typical culture and two varieties below.

B. subtilis (Ehtb.).

Morphology.—Size, 1.5¢-4ux .6u-1.54, commonly forming chains. Spores
are produced in abundance. The Gram stain is positive, and there is no cap-
sule. The size is somewhat variable in different cultures.

Gelatine colony.—A rapidly liquefying colony, with irregularly distributed
granular masses. The appearance of these masses is striking but not uniform,
and hence not characteristic.

Gelatine stab.—Begins to liquefy in one day, crateriform and later stratiform.

Argar streak.—A filiform or spreading, raised, contoured, cretacious, white
growth, frequently wrinkled and dull. In some cultures it is quite dry.

Fermentation tubes.—No acidity, gas, nor closed arm growth, in our cultures,
although Chester says dextrose is made acid.

CLASSIFICATION OF DAIRY BACTERIA. 173

Bouillon.—A sediment, turbidity, and pellicle.
Milk,—Rendered alkaline, curdled and digested at both 20° and 37°

Potato.—A spreading, raised, gray, dry, or moist, luxuriant, wrinkled growth.
Some cultures are yellowish and thin.

Grows well at 20° and 37°. Aerobic.

Variety A.—Differs in the following points from the type. Size, 2.5ux .5u.
Gram stain negative; colony with a radiate pit which is later gyrose; agar
streak shows no wrinkling; potato discolored, a bluish-black.

Variety B.—Size, 3.54x.9u. Gram is negative; liquefaction is so slow that
it hardly occurs on gelatine plate, the colony being round, smooth, raised, en-
tire, translucent, yellowish; a slow infundibuliform liquefaction; acid is produced
in dextrose and saccharose, and a closed arm growth in lactose and saccharose;
on potato the growth is yellow.

Here belong several of the forms of 7ythothrix of Duclaux, (turgidus, fili-
formis, urophalum, ) and also B. Bernensis, found in Emmenthaller cheese.

At this place should be included 7. vergwla of Duclaux, found in cheese,
and B. mesentericus of Fliigge, neither of which is sufficiently described to be
clearly identified, as equivalent to any of the organisms which we have already
described.

B. lactis Cromwelhii n. ». Producing a slimy jelly on potato. This has been
found but once, but its peculiar potato growth demands special recognition.

Morphology.—Size, 1u-.6u. Forming chains. Spores are developed and also
acapsule. Flagella not definitely made out but probably peritrichic.

Gelatine colony.— An opaque colony, in a pit, at first somewhat lobate and
then breaking into opaque granules as the liquefaction increases.

Gelatine stab,—At first a-dry crateriform pit, with later a liquefaction and a
scum,

Agar streak.—Luxuriant, opaque, white, with a thin edge; the whole subse-
quently becoming yellow.

Bouillon.—A sediment, turbidity, and a pellicle, with a tinge of reddish or
brown color. :

Milk.—Rendered alkaline, curdled, and digested at both 20° and 37°; may
digest without curdling. The milk becomes nearly transparent in 12 days. A
yellow scum sometimes forms, but soon sinks to the bottom.

Potato.—A moist, slimy, jelly develops all over the potato, extremely profuse.
White or yellowish-brown color.

Grows at both 20° and 37°. Aerobic.

B. janthinus (Zopt.), violaceus (Macé), A violet bacillus. We have never
found this, but it is said to occur in milk occasionally. Its description is not
complete, as follows:

Morphology.—Size, 2M-5&X .4u-.5h. It forms chains and spores and stains
by the Gram method. Flagella not described.

174 STORRS AGRICULTURAL EXPERIMENT STATION.

Gelatine colony.—A rapid liquefier with a membrane on the surface, which
assumes a violet tinge in some cases.

Gelatine stab,—A rapid liquefier, forming a cloudy liquid and a pellicle;
violet in color.

Agar streak.—A laxuriant growth, at first white and then violet; moist,
wrinkled.

Fermentation tubes.—Not given; probably no acid or gas.

Boutllon.—A turbidity and a slight pellicle; the bouillon becomes violet.

Milk,—The reaction is unchanged, or slightly acid. There is no curdling,
but there forms a violet surface layer.

Potato.—The original needle track is violet, but a dark brown growth covers
the whole surface of the potato. Luxuriant.

Ill. No pigment and no acid in dextrose or other sugars.

B. lactis circulans f. and II, White circulating bacilli, These two varieties
have not been found by us since their original isolation in 1895 as described
previously. The two cultures, found at different times, differ from each other
in the points indicated in brackets, which refer to No. II. Both of them showed
the circulation which is characteristic of this type. Their characters are as
follows:

Morphology.—Size, 1.5u@x 6p. Forming chains.

Gelatine colony.—A protruding bead or in a dry pit; then liquefying and
showing a circulation in the liquid.

Gelatine stab.—Liquefaction slow; a narrow funnel is formed with a dry pit
above, and a rotating axis in the center of the funnel below. [Rotating axis
absent. ]

Agar streak.—Luxuriant, thick, yellowish.

Bouillon.—A sediment, turbidity, and pellicle, but the liquid clears up after
six weeks.

Milk.—Slightly alkaline or unchanged in reaction. Digests without [with]
curdling in 25 days, into a cloudy liquid with a sediment.

Potato.—A rather scanty, thin, watery growth, white. [Reddish brown.]
Potato sometimes discolored and sometimes not.

Grows both at 20° and 37°— Aerobic.

B. aerolactisn.s, A non-acid, gas-producing bacillus. This has been isolated
several times from milk. In one case it was from a sample of old milk that
developed a pleasant, fruity flavor. This culture produced in milk, however, a
strong odor of decay. There are slight variations in the different cultures
which we have isolated, but we regard them as identical. Some of the variations
are indicated by brackets.

Morphology.—Size, 1u-1.2u. x .4u-.8u. No chains have been found. Spores
are produced, sometimes at the middle and sometimes at the end. There is
frequently a capsule and the Gram stain is negative. [Capsule wanting.]

CLASSIFICATION OF DAIRY BACTERIA. r75

Gelatine colony.—A rapidly liquefying colony, cloudy and sometimes showing
a nueleus, but not characteristic.

Gelatine stab.—Begins to liquefy in one day, infundibuliform; complete in
8 to 10 days.

Agar streak.—Luxuriant, capitate, smooth or contoured, gray, moist, com-
monly viscous.

Fermentation tubes.—Grows in the closed arm; produces gas in dextrose and
saccharose and in some cases in lactose. No acidity is developed. [One
culture shows gas in saccharose only. ]

Bouillon.—A sediment, a strong turbidity, and a ring-formed pellicle.

Mitk.—Either unchanged in reaction, or rendered slightly alkaline; is curdled
both at 20° and 37°; is always digested more or less completely and always
shows the presence of gas bubbles. The odor is variable, in some cases being
that of decay and in others that of cheese.

Potato.—Spreading, raised, contoured, gray, moist, luxuriant, with a dis-
colored potato.

Grows at 20° and abundantly at 37° +‘ Facultative anaerobic.

We have isolated this organism from milk from several sources. It is quite
similar to B. megatherium of Du Bary.

B. lactis tetragenesn.s. A rhizoid iquefying bacillus.—The original culture
of this organism was sent me by Weigmann and isolated from cheese. The
characteristics below were determined by us from Weigmann’s culture. Later
we found apparently the same organism in milk here. The points where our
new culture differed from that of Weigmann are indicated in brackets.

Morphology.—A large rod, not forming chains. Size, 34x .74[2.5ux 1.4m].
No spores, no Gram stain, but a capsule is evident.

Gelatine colony.—A rhizoid or proteus-like colony, of large size, slowly lique-
fying. It is not acid in “tmus gelatine.

Gelatine stab.—Begins to liquefy in one day [three days], liquefaction strati-
form. At first an arborescent needle track. [Not arborescent.]

Agar streak.—A filiform, flat or raised, smooth, gray growth, moderately
luxuriant, moist.

Fermentation tubes.—No action upon any sugar bouillon.

Bouillon.—A slight turbidity and a tenacious pellicle is formed, but no sedi_
ment. [Sediment without pellicle. ]

Milk.—No change in reaction, or a slight alkalinity. Milk is curdled at the
bottom, at 37°, and is subsequently slightly digested, with a faint pinkish color.
[Yellowish.]

Potato.—A luxuriant, wide spreading, thin, contoured growth, of a gray
color, with a discolored potato. [Growth is slight. ]

Grows at 20° and 37° [Growth at 37° slight.] Aerobic.

“B. lactis distortus nu. s. This resembles 7y. distortus of Duclaux.
Morphology.—Size, 34x .74. Chains are formed, but no spores or capsules.
The Gram stain is positive.

176 STORRS AGRICULTURAL EXPERIMENT STATION.

Gelatine colony.—A slowly liquefying colony, with a uniformly granular liquid.
On litmus gelatine it is not acid.

Gelatine stab.—A slow, stratiform, liquefaction, with a cloudy liquid and a
scum.

Agar streak.—Filiform, raised, smooth, translucent, white, moist, not
luxuriant.

Fermentation tubes.—No acidity, gas, nor closed arm growth in any sugar
bouillon.

Bouilion.—A turbidity and a thick, wrinkled scum, but no sediment.

Milk.—No change in reaction, or amphoteric; the milk is curdled slowly and
is later digested.

Potato.—A luxuriant, white, much folded, dry or pasty growth.
Grows at 20° and 37°. Aerobic.

B. lactis gelatinosusn.s. A bacillus producing jelly-like milk. One culture
of this organism was isolated from milk here and a second was sent from
Geneva. Where the latter organism differs from ours the differences are indi-
cated by brackets. :

Morphology.—Size, .8u4x.64. [1.84x.64.] No chains, no spores, no cap-
sule, and Gram stain negative.

Gelatine colony.—A round, smooth, white colony, which slowly liquefies
[rapid liquefaction], not characteristic.

Gelatine stab.—Slow liquefier, cratiform, white. [Rapid liquefier. ]

Agar streak,—A filiform, smooth, raised, brownish or cream-colored growth,
luxuriant, moist.

Fermentation tubes.—No acidity, gas, nor closed arm growth in any bouillon.
[Slight acid reaction in lactose and some closed arm growth in saccharose. ]

Bouillon.—A granular sediment, a turbidity and a membranous pellicle.

Mitk.—Rendered acid and curdled, and upon digesting becomes a transparent
jelly.

Potato.—A moderate growth, raised, smooth, brownish with a slight odor.
[Spreading.]

Grows at both 20° and 37°. Aerobic. [Facultative anaerobic. ]

B. lactis tenuis n. ». (Ducl.) Several cultures of xon-acid liguefying bacilli,
with no wrinkling, and no rhizoid colonies have been studied. Here would
belong Zy. tenuis, and Ty. Scaber of Duclaux. We describe two varieties
from our cultures to which we have given the above names, derived from
Duclaux.

Morphology.—A slender rod. Size, 1.24x.54. No chains, no spores, no
Gram stain.

Gelatine colony.—A rapidly liquefying colony, not characteristic.

Gelatine stab.—An arborescent needle growth and a stratiform, later ‘an
infundibuliform, liquefaction, beginning in one day, half liquefied in ten days.

CLASSIFICATION OF DAIRY BACTERIA. 177

Agar streak.—Luxuriant, umbonate, smooth, gray, moist, iridescent.

Fermentation tubes.—Acid, gas, and closed arm growth in dextrose. Closed
arm growth in other bouillons but no acid nor gas.

Bouillon. — A granular sediment, a decided turbidity, and a flocculent
pellicle.

Milk.—Rendered alkaline, digested, curdled, and subsequently digested into
a gelatinous mass at both 20° and 37°, with an unpleasant odor. Later the
odor is of old cheese.

Potato.—A filiform, capitate, smooth, gray growth, not very luxuriant, with
potato discolored.

Grows abundantly at 20° and 37°. Aerobic. Found in Camembert cheese.

Variety A.. .

Morphology.—Size, 2u-3 x .5u-.74. No chains, no spores, a very evident
capsule and Gram stain positive.

Gelatine colony.—A very slow liquefier, not characteristic.

Gelatine stab.—An arborescent needle growth, with crateriform liquefaction,
beginning in one day.

Agar streak.—Filiform, flat, contoured, transparent, gray, luxuriant, moist.

Fermentation tubes.—No gas, acidity, nor closed arm growth in any bouillon,
Saccharose is rendered acid. No other effect in any sugar bouillon.

Boutllon.—A slight sediment is produced, but no other effect.

Milk.—No action on milk.

Potato.—Spreading, thin, smooth, translucent, luxuriant, gray, moist. Potato
discolored.

Grows at 20° and moderately at 37°. Aerobic.

B. lactis plicatus n. s. A spore bearing, liquefying bacillus with a folded
scum. Found in Camembert cheese.

Morphology.—Size, 24x .8u-1.2u. No chains, central spores produced, Gram
stain positive, no capsules.

Gelatine colony.—Rapidly liquefying into a non-characteristic cloudy pit.

Gelatine stab.—At first arborescent; liquefies in one day, stratiform; about
one third of the gelatine liquefies in ten days, with a white folded scum.

Agar streak.—Nodose, capitate, rugose, opaque, white, luxuriant, moist and
wrinkled.

Fermentation tubes. — Dextrose is rendered acid; no gas and no closed arm
growth in any sugar bouillon.

Bouillon.—A granular sediment, a turbidity, and a pellicle forms, which sub-
sequently sinks.

Milk.—Becomes alkaline and curdles at 37°, not at 20°. Is subsequently
digested.

Potato.—Luxuriant, diffused, thin, smooth, of an orange-white color. Potato
is discolored.

Grows at 20° and 37°, but better at 37°. Aerobic.

178 STORRS AGRICULTURAL EXPERIMENT STATION.

B. lactis amberisn. s. A brown fluorescent liquefying bacillus, Found only
in milk from Colchester.

Morphology.—Size, 1m-1.54x 3u-4ph. No chains, central spores, Gram stain
is positive and a capsule is produced. ;

Gelatine colony.—A non-characteristic, rapidly liquefying colony.

Gelatine stab,— Liquefaction begins in one day, infundibuliform; complete in
six days.

Agar streak.—Linear, raised, rugose, translucent, yellowish, moist, wrinkled.
The agar with an amber colored or yellow-green fluorescence.

Fermentation tubes.—Dextrose is acid, the other two sugars alkaline. No
gas, but a very slight growth in closed arm.

Bouillon.—A flocculent sediment, a slight turbidity, but no pellicle.

Milk.—No change in reaction. Curdled in one to two days, and digested
both at 20° and 37°. Digestion is nearly complete in three weeks.

Potato.—Linear, raised, smooth, yellowish, dry, luxuriant; potato discolored.
Grows at 20° and 37°. Facultative anaerobic.

The last two organisms are very closely related and may be identical. The
amber colored fluorescence and the yellow pigment lead us to separate the two,

B. mesentericus fuscusn.s. A brown, spore-bearing acid liguefier. Found
only once in milk, directly from the udder.

Morphology.—A peritrichic bacillus which does not form chains. Size, 1.24-
I.5u¢x.4u-.6u. It stains by Gram method and produces central spores.

Gelatine colony.—A round, convex; smooth, entire colony, of a brownish-red
color. On Litmus gelatine it is acid.

Gelatine stab,—A slow liquefaction, napiform.

Agar streak.—Spreading, thin, rugose, translucent, gray, luxuriant, dull, and
wrinkled,

Fermentation tubes.—Dextrose and saccharose rendered acid, but no gas nor
closed arm growth in any sugar bouillon.

Bouillon,—A slight ‘turbidity is produced, although sometimes even this is
wanting. No sediment, nor pellicle.

Milk.—Rendered slightly alkaline, and curdles in six days at 37°. The milk
is digested, although this power of digestion was lost after long cultivation.

Potato.—A \uxuriant, spreading, thin, rugose growth, of a brown-red color,
wrinkled.

Grows better at 37° than at 20°. Aerobic.

B. lactis vinusn. s. A spore-bearing, acid liguefier. This has been found
but once, in milk in Middletown.

Morphology.—1p-1.2p, x.64. No chains. Spores are produced, no capsules,
and Gram stain is negative.

Gelatine colony.—A very rapidly liquefying colony with a slight granular liquid.
Not characteristic.

CLASSIFICATION OF DAIRY BACTERIA, 179

Gelatine stab.—A needle growth, liquefying in one day, infundibuliform; com-
plete in eight days.

Agar streak.—Scanty, linear, thin, smooth, opaléscent, gray, moist.

Fermentation tubes.—All three sugar bouillons show acidity and growth in
the closed arm,, but no gas..

Bouillon.—An amorphous sediment and a slight turbidity, but no pellicle.

Milk,—Becomes acid, and curdles at 20° and 37°, and subsequently digests,
giving a clear whey, with a vinous odor.

Potato.—Scanty, linear, thin, smooth. Potato not discolored.

Grows at 20° and 37”, but better at 20°. Facultative anaerobic.

Close to this belong the butyric acid organisms of Prazmowski, Hueppe and
Botkin. We have not studied them.

B. lactis Pruchiin. s. A slimy milk, peritrichic bacillus. This peculiar bac-
terium was sent from Geneva. Its remarkable involution forms, and its other
unique characters clearly distinguish it, although it was lost before we had quite
completed our work upon it.

Morphology.—A spore-producing, peritrichic bacillus, with no capsule. It
does not accept the Gram stain. Involution forms, curved, club shaped, and
showing other irregularities, are common.

Gelatine colony.—A rapidly liquefying pit, not characteristic.

Gelatine stab.—Liquefies in one day, stratiform, with a turbid liquid, and a
reddish-yellow sediment. " -

Agar streak.—Round, flat, smooth, opaque, white, luxuriant and viscous.
No fluorescence is seen, although milk is turned green.

Fermentation tubes.—Dextrose bouillon is rendered acid. No gas and no
closed arm growth in any bouillon.

Bouillon.—A viscous sediment, a turbidity, and a flocculent pellicle.

< Milk.—I|s rendered acid, is curdled and digested at both 20° and 37°, witha
slight yellowish color. Later it becomes quite yellow and slimy.

Potato.—Spreading, thin, smooth, brownish, luxuriant. Potato discolored.

Grows at both 20° and 37° Anaerobic.

B. lactis fungiformis n.s. A white, rhizoid, spore-producing bacillus. This
has been found in fresh milk and stable dust several times. The different cul-
tures show slight variations, indicated within brackets.

Morphology.—Size, 34-3.5¢X 1.3u. No chains. Spores are developed and
an evident capsule. The Gram stain is positive.

Gelatine colony.—The colony throws out fibers like a mold, but after two days
this character disappears, the colonies disintegrating into a liquefying pit. On
Litmus gelatine this character is not evident.

Gelatine stab. Liquefaction begins in two days, but never becomes complete.
Infundibuliform.

Agar streak, Filiform, raised, grumose, translucent, porcelain white, dull
[wrinkled].

180 STORRS AGRICULTURAL EXPERIMENT STATION.

Fermentation tubes.—Dextrose is acid, the other two sugar bouillons alkaline.
Growth in closed arm in all cases, and no gas. [Dextrose and saccharose
alkaline; lactose acid. ]

Bouillon.—A sediment, a granular pellicle, and no turbidity. [Turbidity.]

Milk.—Becomes alkaline, curdles and digests with a strong odor. [Acid,
otherwise as above, except that the milk is brownish. ]

Potato.—Luxuriant, rather thick, rough, white.

Grows at 20° and 37°. Facultative anaerobic.

Variety A.—Agrees in all points except that the mold-like colony is not evi-
dent, the colony liquefying very rapidly.

Il. No Pigment, but Acid in Dextrose and other Sugars.

We have found at least three iguefying gas producers. ‘Two of them are
quite similar to B. cloacae, while the other is closely related to it. The char-
acters as made out by us are as follows:

B. lactis cloacaen.s. A gas-producing, liguefying bacillus.

Morphology.—Size, Iu-1.3ux .7@. No chains are formed. The organism
accepts the Gram stain, and has a capsule, but produces no spores.

Gelatine colony.—A slow liquefier, forming a dense granular pit. In Atmus
gelatine there is no acid.

Gelatine stab.— Begins to liquefy in one day and is completely liquefied in
six days. Liquefaction deep infundibuliform.

Agar streak,—Filiform, raised, smooth, cream-white or yellowish, iridescent,
luxuriant.

Fermentation tubes.—All three sugars rendered acid and show growth in the
closed arm. Gas produced in dextrose and saccharose only.

Bouillon.—A sediment, turbidity, and a pellicle.

Milk.—Rendered acid, curdled, and subsequently digested at 20° but not at
37°, showing a yellow color.

Potato.—A scanty, thin, irregular, white growth.

Grows better at 20° than at 37°. Facultative anaerobic.

B. lactis cloacae A.n.s. This was found in milk in this vicinity, but an
almost identical organism was sent us by Weigmann from Kiel, labeled aerog-
enes. It is clearly not that specie, since it liquefies gelatine. Its characters
are as follows:

Morphology.—Size, 1.54 x .5u-.64. Chains are formed in bouillon. It pro-
duces no spores nor capsules, and does not accept the Gram stain.

Gelatine colony.—Round, raised, smooth, grumose, with wavy edge, gray-
white. On Uitmus gelatine a good sized, white, acid colony.

Gelatine stab.—A rapid liquefier, infundibuliform, and showing an abundance
of gas.
Agar streak.—Filiform, flat, smooth, moist, not abundant.

¥ CLASSIFICATION OF DAIRY BACTERIA. 181

Fermentation tubes.—All three sugars show acidity, gas, and closed arm
growth.

Bouillon.—An amorphous sediment, a slight turbidity, and a granular pel-
licle.

Miik.—Is rendered acid and curdled, but with no visible signs of digestion.
It develops a cheesy odor.

Potato.—The growth is very scanty and white; potato discolored.
A second culture of the same differed in showing digestion of the milk and a
more luxuriant growth on potato.

Grows at both 20° and 37.° Facultative anaerobic.

Bacillus ( Proteus) vulgaris (Hauser), This is not uncommon in milk.
The characteristics are as follows:

Morphology.—Size, 1.2u-4 x .6u. Forming long chains. It shows no spores
nor capsules, and does not accept Gram stain.

Gelatine colony.—Very characteristic, showing irregular amoeboid processes,
the so called ‘‘proteus type’.

Gelatine stab,— Begins to liquefy in twelve hours, with liquefaction complete
in a few days. Saccate.

Agar streak.—Luxuriant, moist, slimy, glistening, translucent.

Fermentation tubes.—Not determined, but dextrose is doubtless acid and gas
is produced.

Milk.—Rendered acid and curdles at 20°. Is subsequently digested, becom-
ing yellowish.

Potato,—A luxuriant, yellowish-white and slimy growth.

B. lactis diffususn.s. A pink Bacillus,

Morphology.—A motile rod. Size, 14x .64-.gu. No chains.

Gelatine colony.—Diffuse appearing as a faint cloud made up of microscopic
colonies. To the naked eye it appears like 4 mold; 3mm. in diameter, then
liquefying.

Gelatine stab.—A napiform liquefaction, with a cloudy pink liquid. Below
the surface it is orange-red.

Agar streak.—A luxuriant, pink, moist, smooth growth.

Fermentation tubes.—Probably acid without gas.

Bouillon.—A sediment and a turbidity but no pellicle. The sediment is red.

Milk,—Becomes acid and curdles after several days. No other change.

Potato.—A luxuriant, bright pink growth. No discoloration.

Grows at 20° and 37°. Aerobic. ,

B. lactis cochleatusn. s. A non-gas-producing, peritrichic bacillus, without
spores. This has been observed twice. One culture was sent us from Geneva
by Harding and a second was found in milk in Middletown. The following
description is from the Geneva culture. The points where our own differ from
it are indicated by brackets.

182 STORRS AGRICULTURAL EXPERIMENT STATION.

Morphology.—Size, 1.8u-34x.7¢-.9#. No chains, no spores, no capsules.
Gram stain positive. [Negative.]

Gelatine colony.—A curiously’ lobed colony, cochleate, rather slowly liquefying.
Litmus gelatine is not acid. The colony is quite characteristic. [A simple
lobed, slowly liquefying colony, not cochleate. ]

Gelatine stab.—Begins to liquefy in three days, stratiform.

Agar streak.—Linear or spreading, thin, moist, opaque, white or gray, not
luxuriant. [Yellowish, luxuriant. ]

Fermentation tubes.—Dextrose and saccharose acid, lactose not acid. No gas
nor closed arm growth.

Bouillon.—Sediment and turbidity, but no pellicle.

Milk.—Made alkaline and curdled at 37°. Subsequently digested, with a
prominent odor.

Potato.—Very scanty, white. [Luxuriant, gray-white, with potato dis-
colored. |

Grows better at 37° than at 20°. Aerobic.-

B. lactis Robertiin.s. A non-gas-producing, acid-forming peritrich. Found
only once.

Morphology.—Size, 1.54x.5u-.84. No chains, no spores, no capsule, and
Gram stain negative.

Gelatine colony.—A dense, white colony, very slowly liquefying, Is not acid
on itmus gelatine, but forms a pit colony.

Gelatine stab.— A slow liquefier, stratiform, with a clouded liquid.

Agar streak.—Filiform, thin or thick, smooth, contoured, white, luxuriant.

Fermentation tubes,—Dextrose rendered acid, but no other change in any of
the sugar bouillons.

Bouillon.—A flocculent sediment, a turbidity, and a ring formed pellicle.

Mitk,—Acid and curdled, but without digestion. The milk develops an odor.

Potato.—A luxuriant, thick, moist, white growth. Potato may be discolored.

Grows at 20° and 37°; better at 20°. Aerobic.

ACID GAS PRODUCERS.

The gas-producing Bacteria and Bacilii constitute, with the exception of the
Bact. lactis acidi group, the most important dairy organisms. To this group
belong many of the most mischievous dairy bacteria, Among them are those
that spoil large quantities of cheese by the production of the trouble known as
swelling. Sometimes great quantities of cheese are utterly ruined by the devel-
opment of gas bubbles. The gas bacteria, also, sometimes spoil butter, and

‘they are generally undesirable. Whereas the non-acid-producing bacteria are
commonly the dairyman’s friends, at least so far as relates to butter and cheese
making, the gas-producing bacteria are universally his enemies. In their rela-
tion to milk problems they thus form a group by themselves. In their sys-
tematic relations they belong to different divisions; some of them are Bacteria,

.

CLASSIFICATION OF DAIRY BACTERIA. 183
°
others are Bacilli. But because of their practical association together in dairy
problems, we think it more convenient to consider them all in one group, only
referring to them in their logical place in our scheme of classification. The
other gas producers, clearly not related to these, are described under the differ-
ent groups where they belong.

Two somewhat extensive studies of the acid-producing, rod-shaped’ bacteria
have been made besides our own. One of these was by Harrison, who studied
fifty-six different cultures (Cent. f. Bac. II., XIV. 359, 1905), and the second
by Gruber who carefully studied thirty-seven cultures (Cent. f. Bac. II., XVI.
654, 1906). Although these differ in some slight respects, the general conclusion
from their study is in essential agreement with our own. There exists a long
series of these forms that show slight variations, which grade into each other in
such a way as to make it, at present at least, out of the question to arrange them
all in any logical scheme even if it were worth while. Both Harrison and Gruber
agree that all of these types may be arranged between two extremes, represented
by Bact, lactis aerogenes and B. coli communis. The essential differences of
the extremes are as follows: &. Jactis aerogenes is non-motile, produces no indol,
and has a thick colony on gelatine; B. coli communis is motile, produces indol,
and athin colony. But even these primary characters cross each other more or
less, especially those of indol production and the type of colony, so that they
cannot be regarded as especially characteristic. Indeed, almost any combina-
tion of the above characters as well as others can be found among the many
cultures that have been studied.

Whether, under these conditions, it is worth while to attempt any-classifica-
tion may well be doubted. Harrison does divide them into a series of groups
without, however, implying that his groups have any diagnostic significance.
In doing this he recognizes all the variations he can find between the different
cultures studied, with a result of making a confusing series of types that clearly
have no very great value.

Gruber endeavors further to divide these organisms into groups by their power
of fermenting a long series of carbohydrates, and finds it possible to recognize
four types. But each of the four is found among both the aerogenes and the
coli type, so that the plan is not particularly useful, quite independent of the
fact that no other obsérver has made test with this long list of sugars.

The sum and substance of the matter is, that there is no means at our
command at present by which we may satisfactorily group these types into
definite subdivisions. The plan we have adopted is very simple. We recog-
nize, first, the typical aerogenes type, under which we have referred to a number
of the variations that are known to occur. Then we recognize a type with
flagella, but with the typical luxuriant aerogenes colony. Third, a type with
the typical cof characters but monotrichic, and lastly, the typical co/z form
with its peritrichic flagella.

Gruber states that in the cold types studied by him, the flagella were always
monotrichic and that the name Bacillus should be changed to Pseudomonas.
In our own work we have found both the monotrichic type and the peritrichic
type. The peritrichic type has appeared far more frequently than the mono-’
trichic type, but the latter has been found quite a number of times. Whether
these modifications are the same or really two different types of colt, we do not

184 STORRS AGRICULTURAL EXPERIMENT STATION.
.

venture to determine at present. We think the best course to pursue is to
recognize the two as different, and we have consequently done so in our
classification. :

The following classification is based upon the work of Harrison and Gruber,
with the aid of such additional data as we have ourselves obtained.

BACT, AEROGENES TYPE,

This organism has appeared in literature under a long list of names. The
organisms of the following list appear to be identical with each other and are of
this type. 28. pyogenes (Albarran), Bact. aceticum (Baginsky), Bact. theloideum
(Gassner), Bact. udiguitous (Jordan), Bact. candicans (Frankland), Bact. zur-
nianum (List), B. capsulatus (Smith), B. chologens (Stern), B. acidi laevolacticé
(Kozai). It is by no means certain that these organisms are identical, but the
descriptions given of them agree so closely as to lead to the conclusion that
they are essentially the same. The general characters of this type are as
follows:

Bact. lactis aerogenes (Esch.). The non-motile, acid, gas producers.

Morphology.—Size, 1.4u-5@x I¢-1.5¢. There are no chains, no spores, and
no flagella. A capsule is frequently found and the Gram stain is variable.

Gelatine colony.—Large colonies, 2 mm. in diameter, thick, round, smooth,
moist, sometimes viscous. On Atmus gelatine they are essentially the same,
and very strongly acid. They frequently show bubbles of gas under the sur-
face of the gelatine.

Gelatine stab.—A good needle growth, and a thick white surface. If sugar
is present, gas bubbles may appear.

Agar streak,—Luxuriant, moist, gray-white, smooth.

Fermentation tubes.—All sugar bouillons show acidity, closed arm growth,
and an abundance of gas.

Bouillion.—A turbidity and a sediment, and commonly a pellicle.

Milk.—Becomes strongly acid, and curdles, bubbles of gas being commonly
evident. This curd is usually very different in appearance from that produced
by Bact. lactic acidi.

Potato.—Luxuriant, of a dirty-white to straw color.

Grows at both 20° and 37°, but better at 37°. Aerobic. Indol is not
produced.

Twenty-three of Harrison's organisms belong to this general type, and we
have ourselves found it extremely common.

Among the varieties of this typical form which appear among the many
cultures studied, we make special reference to the following:

Variety A.—Produces indol. Among the different cultures belonging here
some produce the typical thick, aerogenes colony, and some the thinner um-
bonate colony with a smooth center.

Variety B, Produces no indol, but has a thick colony more like that of coli.
Among them are some which produce no acidity in saccharose, and do not
curdle milk, and others that ferment all of the ordinarily used sugars.

CLASSIFICATION OF DAIRY BACTERIA. 185

Varieties C.and D, These two differ from common aerogenes bacteria in
making mz/k bitter. They also show some other rather peculiar characters, al-
though they plainly belong to the aerogenes type. Variety C was isolated by
ourselves from milk in Connecticut and differs from the type in the following
points: the litmus gelatine is not acid; a brown fluorescence is produced on
agar; dextrose is acid, but no gas is produced; milk is curdled at 37° only, and
is very bitter; potato growth is very luxuriant, white, and the potato is dis-
colored.

Variety D, sent us by Harding, produces gas in dextrose only, but all three
sugar bouillons become slimy; there is no closed arm growth, no pellicle on
bouillon; milk bitter; potato growth is luxuriant and the potato is discolored.

THE COLI COMMUNIS TYPE,

The rest of the gas producers found in milk are flagellates. Among them
we recognize some with thick colonies, like aerogenes, and others with thinner,
spreading colonies, of the co# type. In most cases the flagella are peritrichic,
but there is one type found by us several times that is monotrichic. Beginning
first with the types most similar to aerogenes, we recognize the following
varieties:

B. coli aerogenes n. 8.

Morphology.—Size, Iu-3u x Iu-1.4@. There are no chains nor spores; Gram
stain is negative, and the bacilli are peritrichic.

Gelatine colony.—Prominent, thick, moist, smooth, large, surface colonies.

Gelatine stab.—A needle growth, and a thick, white surface growth.

Agar streak.—Filiform, raised, smooth, opaque, cream-white to brown.

Lrermentation tubes.—All three sugars are rendered acid, gas is produced, and
there is growth in the closed arm. The amount of gas is not very great.

Bouillon.—A sediment and turbidity, and usually a pellicle.

Milk.—Rendered strongly acid and curdled with gas bubbles.

Potato.— White to straw color, luxuriant.

Grows at both 20° and 37°, better at 37°. Aerobic. Indol is produced.

Some twenty of Harrison’s organisms would belong here. They differ among
themselves very much in their gelatine colonies, and somewhat in their growth
on agar. There are also differences in their powers of fermenting different
sugars. We do not see in these points any good reasons for separating them.
One variety, however, may be properly recognized in accordance with the plan
adopted above.

Variety A.—Produces no indol.
Here apparently belongs 8. Schaffert.

B. coli communis (Esch.) This differs from the last chiefly in producing a
thinner colony on gelatine, which is umbonate, and has a granular, lobate edge.
Its agar growth is smooth and white. Upon potato it does not usually grow
luxuriantly, and indol is produced.

186 STORRS AGRICULTURAL EXPERIMENT STATION.

&. coli is very common in milk, as would be expected from the frequency of
fecal contamination. The distinction between this and the last type is not
sharp, and perhaps should not be recognized as marking separate types. The
organisms which we have called aerocoli seem to be in a measure intermediate
between aerogenes and coh, a fact further suggested by the study of some cul-
tures sent me by Weigmann and labeled aerogenes. These were distinctly motile
and peritrichic when studied by us, suggesting that either the presence of fla-
gella has not always been regarded as diagnostic for separating coli and aero-
genes, or that a non-flagellate type may later: develop flagelli. The typical
characters of this group are as follows:

B. coli communis.

Morphology.—Size, 1v-1.6u x .4u-Iu. No chains, no spores, no capsules.
Gram stain negative. Flagella peritrichic. :

Gelatine colony.—A rather thin, spreading colony, umbonate, with a smooth
center, granular edge, lobate. Litmus gelatine shows a dense white colony,
which is decidedly acid, and may show gas bubbles.

Gelatine sta#—A filiform needle growth, with a spreading, moderately thick
surface growth.

Agar streak,—Filiform, raised, smooth, sometimes lobate, opaque, white,
moist.

Fermentation tubes,—All three sugars are rendered acid and develop gas.
They also show growth in closed arm. The amount of gas is somewhat varia-
ble, and the proportion of hydrogen and carbon dioxide is approximately two
to one, but quite variable.

Bouillon.—An abundant turbidity, a sediment, and’ commonly a ring-formed
pellicle.

Milk.-—Is rendered acid and curdled. The curdling is not absolutely con-
stant, however. Sometimes the milk does not curdle till after it is boiled.
There is never any digestion, but a whey may separate from the curd.

Potato.—A moderate, smooth, gray-white growth, sometimes luxuriant.

Grows both at 20° and 37°, but better at 37°. Aerobic. Indol is produced.

Variety A, Agrees with the type except that it fails to produce gas in sac-
charose or lactose.

Variety B. This differs from the typical coli chiefly in its action on milk,
which it turns acid with a viscous coagulum. This is extremely slimy. Its
colony is umbonate and tenacious, and can only be removed from the gelatine
as awhole. In other respects it agrees with coli. Harrison regards it as
aerogenes, but since it is motile, it must be grouped here,

Ps. coi communis n. s. A gas-producing, non-liquefying Pseudomonas.
This organism and two sub-varieties were all found in cheese made at this
place. They are probably physiological varieties of the same organism. Vari-
eties C and D came from the same colony and are interesting, therefore,. as
showing a possibility of physiological variations from the same culture.

Morphology.—Size, 1u-1.54 x .84-.94. No spores, no chains, no capsules;
Gram stain negative.

CLASSIFICATION OF DAIRY BACTERIA. 187

Gelatine colony.—A round, thick, smooth, or contoured, auriculate colony, of
agray color. On ditmwus gelatine a large surface colony is produced, strongly
acid, and with a gas bubble. Resembles B. dactis aerogenes. In some cultures
it is thinner and of the coli type.

Gelatine stab.—A filiform growth, and an umbonate surface, with a bluish
sheen over the surface.

Agar streak.—Not luxuriant, linear, raised, smooth, gray.

Fermentation tubes.—Acidity, gas, and closed arm growth in all three sugar
bouillons.

Bouillon.—A sediment, decided turbidity, and a flocculent pellicle.

Milk.—Rendered acid and curdled at 37°. No digestion, but a prominent
odor.

Potato,—Moderately luxuriant, thin, spreading, gray.

Grows better at 20° than at 37° Facultative anaerobic.

Variety A differs from the above in the following points:

Variety A. Size, ux .5u. Flagellum very long. Gelatine stab, spreads
from needle track below surface. Agar, luxuriant. Milk, curdled.

Variety B. Size, 1.24x .5u, Gelatine colony, thin, transparent. Not acid
on litmus gelatine. No acidity nor gas in lactose or saccharose, but showing
closed arm growth. Potato, growth thin, cream-white.

These organisms are in nearly every respect identical with B. coli communis,
except that they have one flagellum instead of many. This flagellum is very
long and characteristic.

CLASSIFICATION OF DAIRY BACTERIA.

Bacteria that are spherical in form, Coccaceae.,
Bacteria that are rod-shaped, Bacilliaceae.
Coccaceae.

Coccus forms that do not liquefy gelatine, 108, 188

Coccus forms that liquefy gelatine, II7, 189

Sarcina type, 124

Bacilliaceae.

Non-flagellate rods (Bacterium), I.

Flagellate rods (Bacillus) II.

‘Bacteria and Bacilli which produce acid and gas, - 182
I. Bacteria that liquefy gelatine, 136, I90

Bacteria that do not liquefy gelatine, 125, 189

II. Bacilli with flagella over the whole body (Peritrichic), 164
Bacilli with a single polar flagellum (Monotrichic), 150, 191
Bacilli with a tuft of flagellaat theend (Lophotrichic), 159,191

STORRS AGRICULTURAL EXPERIMENT STATION.

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ay YSIMOTIAA = MOTTE. umolg pet quaosaiony iq
a | l | | l
ul
‘aSOULXIC NI GV ‘aSOULXIC NI GIoy LON

‘2090 SughfanbrT :

STORRS AGRICULTURAL EXPERIMENT STATION.

190

°

6br ‘d ‘wnusew
gbi d‘nunos = gh -d ‘wnyeond
| |

Peploj jou ‘Too —papjoy Auojoo Ler -d
| | : Lei -d ‘umnyeoiny
fbi ‘d ‘suaosaiony ‘slAoiq -OOSIA
| |
juadsaiony JON juadsaiony Aull[s JON eke

| | | |
| ]

€br ‘d ‘nuojysy
zbr cd ‘usanquny

ebi ‘d ‘wnajny itr -d

Pie!-400 pany):. PRE PND Ebr -d'ungye rbr-d‘nypeysseyy ‘wunssynurm = obi “d ‘wnuoiy9
6bz ‘d ‘wmnnooog | | | | | |
| | ayvUM Mood aduzIC asuviO MOT[e4 UOWAT
a on Pipes AA | | | |
. ‘
5 a 0) oobi ‘ding ofr ‘d‘wniqni 6€1 ‘d ‘sauasoiyyA19
gbr “d ‘souadosaenbyy MOTIPA pey Mois YUlg jusosaiony yuULg
| Str -d ‘aevovola | ieee |
iz
sed ON sey gfr‘d‘umasuayg Lei‘d L€r‘d ‘yy sussseioqie gti 'd
| | E ; Str -d bbi-d gerd ‘nuesiysiy ‘umjeounsy LE1 “d‘swaiojyyy ‘wnjewoiy
[ Str ‘d ‘umyeqo] ‘umasrjza19 ‘losnur

ary MN | | | °
ysimorjad y aBueiQ = plozlYY ION ploziyy
| | | |
| |

saiods on saiodg
| : |
‘ |

“GSOULXaAd NI ALIGIOV

soiods on

| |
| aXT] aovyins
oNslejoeIeyo JON xXBIyjUy ssejs punoiry MOTI
| | | |
| |

saiods

“USOULXAC] NI ALIGIOV ON

‘wnigIvg —spor Surtfanby ‘ajo -uony

191

CLASSIFICATION OF DAIRY BACTERIA.

€gx ‘d ‘snapropnsour €gt “d ‘[I] susoseiony
ay MA used zgid ‘snujssnuya =19T “d “y] suaosa10ny
| zgi ‘d ‘[] susosaiony igi ‘d ‘iusisy

gsi ‘d ‘squaStwuesaid | =
6S1‘d ‘enueis gS ‘d ty suewea | \
| | : uaai3 uMmoiq
pa[pano 10N paypino 3A gousosaiony ON jugosel0ny yusose10ny |
| | | | |
6s1 ‘d
LS1 ‘d ‘susoseion] ogi “d ogi ‘d ‘snuedsouds
| ogi ‘d ‘stiepnosjour ‘snynuiu “snadeAl[O |
‘ony ON qusoseiony J | \ \ umoiq
| | anya MOTPA —- W224) yuaossiony
ie | enna | | | |
a
LS1 ‘d ‘sauesoi0e *ONIAATOOIT “ONIATTNOIT-NON
|
se3 ON se ‘ypowg, I0y214qoy Go]
| |
gsi ‘d gst‘d SS1 ‘d ‘eur zS1 ‘d ‘sisojno1eqny “sd «= 1S1 ‘d ‘stwIOsY
‘ane ‘taeqoy «SSI -d ‘enum $$$r ‘d ‘ej10}U09 : | |
| | | | ard AA MONA
MOTPA aya ‘“d4jod ON wuosidfjog | |
| | | | est :d
‘onoejodes ZS “d ‘sIpliA 1$1 ‘d ‘wuaysq OST -d ‘eaine
ploe jou pre uMmoiq usai3
saiods ON saiods AINA AW ‘ONY ION JeosesON] yuaosei0ny I
| A | | | | | ost ‘d
psi ‘d ‘eis esx ‘d Sgr ‘d ‘109 ‘“qumo72|PPLINL
pS1 ‘d ‘sejoingy “eueue | |
juaosoiony JON UMOIG “AON umolg se3 ON set) sed ON sey)
| | | | | ; | \
| | |
poy ple joN —-2s01}.Kep Ul PPV 2 poe ION

| | | |

“ONIAMANOLT % *ONIATINOIT-NON =
spuomopnasd =Ypog IYILAJOUDPY

STORRS AGRICULTURAL EXPERIMENT STATION.

‘192

ggi “d ‘avtajuishp
Lor ‘d ‘snyeojns gor *d ‘suaroejiqni

seiods on saiods ¥
| | : Sgr ‘d ‘t19ysayojo)
Lot ‘d ‘ovnesesy | por ‘d ‘exueZ por ‘d ‘snosejoisiu
Ploe ION pre ATW RTT ION yoriq 19f
| | pgi “d ‘souados1ae . ‘| |
| ggi ‘d ‘sneiy19 Sgr ‘d ‘too Sg ‘d ‘sninqou |
ses ou ‘uy AA MOTPPA sey ploziys jou Auojos) proziys Auojod
| | | |
‘aIy ‘dInV ON

‘ypwug ny nspdag Surtfanbry-uony

193

CLASSIFICATION OF DAIRY BACTERIA.

Be YU sioyue
g4i ‘d ‘snosny LL1 ‘d ‘snyeoryd

| |
Jayanby Mog s1eyanbry pide yy

6L1d‘nyonig gli ‘d‘snura | |
| | |
Awys 4TH PHe ATLA =PpPpe jou ATLA

gL1‘d‘smuo} igi -d Tg1‘d ‘suespna
zgi‘d ‘iqieqoy ‘snsnyip ogi ‘d ‘y ‘avovoja IL. wall |
Igt ‘d ‘snjeayyooo | ogi ‘d ‘avovoja § = 641 “d ‘syuopisuny |
se3 ON yd sep Prozryt "[OD Prozry1 jou "JO
| | | | |
| |
soiods oN soiods
| |
gli ‘d ty ‘suey gf1‘d ‘snsoupees
|
ax Atial JON mas THN
ie
SLi ‘d ‘snqsoystp Sr ‘d ‘sauade1j9}

, | | 641 ‘d ‘stwiossuny

P2TFULIM JON = PeAUUAA prloziyyY zL1 ‘d ‘saprookur

| | | 2L1 ‘d ‘iyjaMpoig 1L1 ‘d ‘saprozry1

‘d 6 ‘d be
yh PLi ‘d ‘sijoejo1se a ca es aye ‘a pease nde
ee oN ig Epa ION ne
| bid ee €Lr-d ee 4 ‘snsnyip €L1 ‘d ar i
Sat AA YSIMoy]a A JOLOLA 4d SU AA YSImoy[o A
| | | | | |
| |
soiods oN seiods
| |
|
PPV Pre ION

oli‘d oft ‘d‘*ay O6g1'd ‘nuoswiepy = ggi ‘d ‘iqna
‘ra31u suasseiony 691 ‘d ‘snuoijio ggt‘d ‘snsorsipoid
ysraorad | | | |
Jo ayy YR Wweoseson,y — AojjeA-uowaT PPy

| | | | |

-wngwg aynaitsay Surtf[anbrvT

€zx ‘d ‘snpiqqe sroey “ww |**|—|=/+]°°|+)-+/+/—-|4+]+/+]+]—-|-|-|-]+/-|-;-]+]-] +/+ Fl- =~ ‘ooeze'11z
zzi -d ‘snsogni sigoey “yy | |—}-E ||] yy] +] -}+]—-]—]—-|-]4)+]+)4+)/4+)/4/4/4iCl-|-|+]- > > (goeeetre
ax ‘d ‘snaquesid syyoey we f° ||] ye] ++] +)—|—|-+|—|+|—|—]}-}-|4+]4+ —|—|+ y+ +) - - - ‘gowze'11z
121 ‘d ‘suerea soe sy ft {—}+ i+] | + i+] 4+)—]4+}—|4+|—|—|—}—}—] 4 4] } 4+] FI) +] 4+]—-|-}-| F] > s - ‘Gosee lez
oz1 ‘d ‘suassaiong sioey “yy | |}—)+)-+] °° }+/4/+/4—|+/— —|+)+/—/—}+]+}+ —|—| - - - ‘IO€ES 12s
ozi ‘d ‘snqje siyoey f+ }—i—}o}-+ i+ }+)—}+)+)+)+ —|—|+/—|—|—] + | —} +] +] -J-]J—- J] - - - ‘SofE 11s
611 -d ‘y snaine stjoey ‘yy f°} J—/-+ 1°} + )+)/4+/—]4+]+/—|+/-|— +)+]/—|—|—| +} +}+ - - - ‘SofCe11z
611 ‘d ‘snuussynurw soe] "wt" * +e] el) +]4/—)+)—]—|-}]—] +] +--+} } 4] J - - - ‘GofEE11z
gir “d ‘aso0y sioey gs ffi] + )+)+f+) 4] }-f—}-i-|-]4}4+)-i-J4]-|+/#I-l-l+}-|- > > ‘sob€e-rre
git ‘d ‘snesyo sroep “sg |**|—l—j—} | F444 |—|— | — |] | J] 4] 4 JI] 44+} +i +i} +i] - - - ‘SofEEIIZ
L11 -d ‘snuosyo susey “we | *|—|—|—]*|—|+)-+]+]+ —|—-|—|—|—|+}4+]-J—|4+]4+]+}4l-j]-|-|-]- - > ‘goeeerrz
£11 ‘d ‘snpiqny story “yy f°] + |—|—}e |] 4+ }4+}+]—|—}—|+)—}—}—-|-]+]+)—|-]+]—|+} °° —|- - - ‘Lo€€e11z
Lui ‘d ‘sauasoiyyAs9 syoey “yy fo ||} fs |i) 4]-4+]4+)—j+ —|—|—]+}+}+]/+)+)—)+]--}-j]-]-|-} - - - ‘IO€EE 11s
bri ‘d ‘snonoey story *s |**|—/+}+/— +{—|—|+|—-|—|—|-| —| + —|+/—}+)+]/—|—|+/—-!} - : - ‘O0GZZ ‘71%
gr ‘d ‘tproe sory ‘yw |: -|—|=|/—|—|—|-|4+] -|44+]4+|-]— apf ded ks oh | | se) - - ‘00%zz'z1z
gir ‘d ‘sears soe, ‘we f+ *|—|+)+/—|—|—] +] —|—| + ]—|- |] —|—I- | +] | - || +] —-|-}+-|-}-|-—-|4)} > - - ‘00zZz'Z1z
S11 ‘d “yy pue ‘y snoyoe, suoey “sg f+ + |—|F/—|—|—|—} + ]—|—} +] 4] —|— |] + J] —| 4] 4] 4 ]—} +] F} > : - SOOEES Zz
€11 ‘d ‘snsoosia syjoey ‘Ss |**|—|+)+/— +i—|+} +} + |—}|—|—}—}—} 4 || 4] —J—|—]+)4+] - - - '00%2e'°Z1z
€xx ‘d ‘snaime syoel “S| *|—| | |—|—|—}+4 J+] 4+] 4) 4+] || jj Hy ey] y+} —-}-] +] +] - = - ‘gozzz'ele%
z11 ‘d ‘sname sroey “Jl |**|—|+) = +] +) +] +)+)—-j— ee ise te |= (1 | = - ‘Gousz 1s
arr ‘d ‘snaqny soy *g fo +|—l+) +|—|—|-]+|-|+]+]4+/—]—|-|-|-|+]+|-|-]+|-|+]+/-|-]+f=]- > > ‘goeeerer2
ort ‘d ‘10joo1ss9a “2 |**)—|F —|—|+]-|-]+)+|-l-|-|-|-]+ FI |-|+|-|+)4/-|-|+]F]- > > ‘ootee ere
or1 ‘d ‘suassaioqie sioey “ |* *|—|—|—|— +/—|—}+/+/—|-—|—}+-|— —|—|+-|+]+]-°}-j- - - - ‘OoESE Zz
6or ‘d ‘9 snsoosta sijoe] “y |** —|—| +) +)+]4+)+)/—-|-—|— +)—|/—|—|+]—|+]+/—j]-|—|-] - - - ‘OOEEE Z1z
6or “d ‘snaey sioe] “IW fat me lem ame Red feat nN Qa el eal ee el ied ee ee a Ol em ee lc OO Ce Oe Gnu i = - ‘goeeeziz
gor ‘d ‘snaxz19 siq9R] "I —|4J—J~)]— J] 4] |} }— JJ] | J] ] 4] 4) +] 4) -] -— - = - ‘So€EE-z1z
gor ‘d ‘snaoesor soe] “IW + J—|—J— Je JJ J J I J] 4] 4} 4+] -}— = - - ‘LOEEE*Z1z%
7) Q nD nn wD
glee /e £1 e)8]9|81 2) ¥] 2] 2 2/2 2 zigiSisteieiel2\s|aigigie
O/B /A A SIAR SS sl AalselalalalSia/e/slal |g |B Sel Blals| es
“Vel |ajelFel@/o)elm(g/2/Flale/e/S/S/F) jal /BlalelFiels
ey yal [Piste eels er /eis|sie) |e) |S/ ei g
a) | IsfPlP isis gi Riga; | [| "|e g
“AWN : 8 4/4 2 Plo) e a 3 ‘WaSWAN ano0¥9
Pye |e 4
: : Z
cant | SB) | AS | cava eo | re8v | ure
“IVOINAHD-OIg "SAUNLVAY IVAYOLTNAD “ADOTOHAYOW

‘avaIvIIO)V —"I WIAV I,

g€1 ‘d ‘q iploe sox “oe |**|—|-+ aE cal ie! cared fad cs —\+)4+)+\— F\— = - ‘oo€ze"z1z
g€r “d ‘3 ippe soe “joerg |**|—)+/-+|— + Ble =F —|+)+)/+i+ +|— < - ‘00tZe'Z1z
S€x ‘d ‘g iptoe stjoey “yoeg [**|—)+|— +)—J]—} ++ ee es Oe eu far ae (em . = “gozectetz
ver ‘d ‘iproe sory ‘yoegq |**|—|/+|+/— +i—|—] + fe es |e Se | a oe . - ‘oozzz'ziz
€€r ‘d ‘wnymbiqn syovy yoeg |**|—/+/+ ers yg as =| ese] e] 0 +4 i = MaggeErere
z€x ‘d ‘ipioe vou syoel “yor |* "| + = +] 0) }-+]-+]—-|— += |—J—]4) 4+] FI -|—] 4] + - = ‘ooze z1z
z€r ‘d ‘wusisy sioey ‘yoeg |°*|—!+|—|—|-+]—|+]°*|+)4+-)+ —|—|—|+| F)-|-|+|—|+|— —|— - = ‘o0zzz'zIz
z€r vd “yop ayes “yorg |**)—|—|—|—|—] F J+) [+] +) +] F]— —|+)#|-J]-'+|-|+ |= - - = ‘ove zz
rér‘d ‘umyjuexudz soe] “yoeg |**|—|-+|—|— +I] AIA] |||] 4 FJ] 4] — - - ‘00%ZZ"Z1z
of1 ‘d ‘yy snaine siyoey ‘yoeg [°° | F/—|—|—|—|—} F} | —]| +] +|—J]—|—|-J]—-|+-]+ bie dl Fes ea RE . = WGoesersre
oft d ‘sisuouayeo story “yoeg [°° |—|F]F Slee) ela le eee ee e - ‘Cote zit
6z1 ‘cd ‘stsuapns syjoe, “yoeg |**|—|+]+]/—|—/—|+]--|—}+) #/—|-—}]—|-}-}+ Rill aie Briel ere len - - ‘Lozes'ziz
gzr ‘d ‘nuuod sje] “yoeg |**|-+|— SSG SS eer | ia] ae ae eS ee = - - ‘Oo€€e*ziz
ger ‘d ‘a iproe sory joerg |**|—|—|— —|+|—|—|+ zl fe reli ar (eal peek en ae x - ‘Oo€EE*zIz
gzr ‘d ‘wnsoosia siovy “yoeg |**)-+ Sd (oer em. bee] —|-|-|4+)—|—|-}+| F]4+]- +/+ = - ‘Oo€£€*ziz
der ‘d ‘7 ‘suaosasoqie siaey “yoeg |**|/+)—|— +] + f—]+}+)—]—} —|—}—] +] — sr a ro 2 - ‘oof€€'ziz
Lex -d ‘unpprojaoAur story “yoeg |**|—|F)—|—|-|—] "|| +] +|-|-|-]+|-|-|* +)—|+) F |-|-|4+)— - ~ ‘00€€E*z1z
gerd ‘yy ‘was syoel joeg |**|—|/—|— i a lal ol ase Sooed i freed fatal Gao feed (Gand aL fal led et Peel fl = - ‘SO€EE-z1z
gzi‘d ‘*y wnaine soe “joerg |**|—|—|— ape ro Fmt Pe) Ea PPA PF re or |e : - ‘Q0€€E‘z1z
Sex ‘d ‘stuowyes sijoe] ‘youg +|—j— [epee pall of [ese | Jed poe | |e - + "LOSE 2tz
ot oaEIKe) Oo) 410 fe) n|D alo
E RlE/E ela lel clei el alata [slelelele ls ale ele glelelglele
PEP SBE ES alo/sle/s)slale|Sia/6/e/B) gl Bela |el alee
B] |BlelPle sles) ®|8/2) Peles elslE| je) |Blalgye leis
te “48 lela | 199 Jaq elo |) 8] le . Z| eB \° i}
5 SPIT ata 2 RB) a |* : “|B 9g
‘aNVN B 2 4 ‘ | |e 3 im e ‘WaHEWON dNnoUD
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sau | Sbee | | Soa | SSS | carta eo | se8v | moe
“TVOINAHO-O1g "‘SAUOLVEY TWUNLINAD “AOOTOHAUIOW

“SuitfanhrT-uopy ‘wnisavg—'t AIA I,

6b -d ‘wnnoov0g soe “yoeg |**|—|-+]—|0}— |) Fy 44+ —|—|+)- +i+|+)+ —|F) - - - ‘o0€€e"11z
6bz ‘d ‘wnusew syoey “joeg f°} ||] | y+] 4] rye] e+ +|/—j|-|— —|+)+]4+]+i—j—|4j)+)- - > ‘oo8eec11z
gbr ‘d ‘nuri0y sie yowg [°°] J—|F] ||] y+] +] 4p] +|—|—|—|+]+}+]+ —|Fl- > > ‘oozes 11z
gbi ‘d ‘wnzeord stor] “joeg see ole df} ]+e|4l+]+]4]—l+] éJ- — —|+)/—|+]#/—|—}+/-] - - - ‘00%ZZ' 12%
Lox ‘d ‘suaosaiong sory ‘joerg [**)+)+]—]°}+]-4) Ff] Fy} éjAi—l— yy 4} I 7] > i - ‘1OZes* 11%
Lord ‘staaiq story “qoegq f° * ||] |) 4} | +] 4] = —|+)/—|—} Fj—|-— ||} - - - ‘00zZz" 1Zz
gti -d ‘umnyzeoinj-oosia sijoel “Jorg {opted |ele ye] 4] 4 I-A] 4A] il] Fl] Fp] |] - - - '0%%G" 11%
gti ‘d ‘souadossenbi] sory joeg |** +l} 4+ I+E]|+)/—|—|-—|-|— oe —j|+)+/+/—|—|-—j-|-}_- - - ‘OOIEI 12%
Shr ‘d ‘avovojo sijoeR] “Rg soft fy 4] 4+] 4] 4] ]— + +)+j)—|+}— —|—| - - - ‘OOIII*11Z
Shr -d ‘wnyeqoy soe] “yoeg [**|—|+]/—]*}A I+] Fy pF] +} —-) +/+/—|—|+)+]+]+\/-—l— - - - ‘Oo€e1zz
Str -d ‘umaoejaio story “yoregq |**|-+|—)+] 0} )A) A) 4AY 4A) 4] +] FI) -}— +|—|—|—]4+)+|4+)+i—|+)/—|4+] - - ‘oozfe IZ
bri ‘d ‘iosnui sioe] “joerg {| fee |e] le el] 4 ]—l—J—} 4 J }—|-]4]—] +] |4+]4)-) 4+) 4] * - - - ‘oo€zz 111
Ebr ‘d ‘wnqye siyor] ‘joerg «flee ete] 4] —|—-] 4 |---| |] 4 |} EI] 7 - - ‘Oo€EE*11z
€r1 -d ‘tuojysy sie] ‘joVg fff ete] ete] e |] ] iy I) y— | it] —|-+]| - - - ‘SofEl*1zz
zbi-d ‘unayny story oR fe} J—te fe ep }] JIE} —AJA il] Py 44 J -|- IIT] 7 - - ‘SofEE'I1z
zbi -d ‘nsinquiy suoey ‘joerg |**|+j—|—] fF eye] +/+/—|—]-—|-]+|-— —|—| - - - ‘oofe£11z
rbr-d ‘iyeysiey stoel “yoeg f°} |—|—]°|-F 4] 4+] +/— —|+)+)—|—|+]+)+]— —!+] - - - ‘So€EE*11z
ib1 ‘d ‘wnunssiyynuriw stjoel ‘joeg [°° |+)|—j—}"*|—|+|F ofa] eff | |] +] J IJ EY] FEL] ]—] +I] > - - ‘Go€ee* iz
Ibi ‘d ‘vy stuoszo sje “joeg fee] |e te} 4p |] 4]—}4+]-]4+)/—j—} 4] 4+} 4)-| +]—|4+ +/—|+4]+i+] - - - ‘Goffe 111
obi ‘d ‘wnuoiqo sis, “yoeg |°*)-+/—| F pete eye |) ]—}4+]—|—l—Aj—} +] 4 y+] -}4}]-}4+}+ +/i—| - - - ‘GofEe*11z
obi ‘d ‘ting sory “jorg fl pe ]—te }—t el }-] +I] 4 J-IAJ— IF FIA] IAFF] 7 - - ‘Lo€€e11z
oti ‘d ‘umiqni siyoey ‘yoeg |°*|-+)—}+] + )+y+ ++) +)/— —) +/+] +i+]4j—l—}}—j—i-}—-| - - - ‘GofEE11z
6€1 ‘d ‘sauaZoayyAra syjoey “yowgq |**|—|—|+] 0) + )Ay yy +] +i 4+ — |) +} + |i] He] +] 4] il} o> - - ‘SofEl1zz
gf ‘d ‘wmaauey sory “org Peto Pteye cf} lelei ty +)4i— —|—|— | + J|—)|—} Fe] +] + j—}4+)—i+] - - - ‘Oofee 111
gtr ‘d ‘nuesiyory soe] ‘joeg eff ede |i pt |p 4]—l4|—l-]—|—j#}—]4)-y +4] 4l-}-]4+}4\7] > i - ‘ooeee 111
L€x ‘d ‘winyzeouna} styoe] ‘yoeg fefefaeye |e ytd 4p )+)—-]4-]—l-—|-l-|—-|-]-) Jy} -} 4] 4] F 2 : - ‘oo€ee* 111
LE1 ‘d “TJ suadsasoqie sijoey “joe fff Pepe lbp} ]4]—]4]-]4+]4]—|—|-—|]4) 4) Fy +l + +/+i+] - - - ‘oozez 111
gf1 ‘d ‘umnjeworyo ste] “JOR_ +/—]+ ley +l ]4+}+]/—}+)]—}+ +l J+ |i y+} - - ‘Goffe Lit
oy loun--Ine) > ols} oO} y ° Din n|O
BRB Bae eeaBeeeeee
S/S[AS/RICElS SBIR Sle ale iS |s/8/g/E)F le B/Zie (e/g |o |e
B) (BlelPigl@ieele ai2|*lalalsieiB/2) (e|° |Blalaie|? [a
o Pla e/2ItlBlo Bm 218 1a) 2 B Ble |S o
. 4 ® og mm} oO [¥ fam tq] 4
=] ow | Qt e 4 ° RIB oO}: f Ss fe Si
B % 3/3 & 0 || : <
‘aWVN " : S| a zi 5 2. a ‘aaaWwoN anowy
E £ :
ont | Sabye| aca TS | ceria. | se8v | wor
“IVOINAHO-O1g ‘SAUNLVAA IWUNALTIAD *ADOIOHAYOW

SurtfanhrT ‘wuniuagovg—e WIV,

“plow jou ‘seg x

6S1 ‘d ‘enuesd sox “sq |**|+}—j—]**|—|+)+|—|—|+]+]+|-|\—|-|-|+ |J—)-|—-]+-]+]+)—|+]4+/-|-] - > ‘ooeee rrr
gsi ‘d ‘tased souasimesaid ‘ov sq [°° }—|+)+] ]4)4I4L yy ye a asa alll Ul As - ‘006 2° 121
gsr ‘d ‘suepea-syoey ‘sq f° }—|+yt] |) 4} |p i+ 4] 4] ] | 4-4 Fi] 4] > - ‘oof€e"11z
LSr ‘d ‘suaosaiony sioey “sq |**|-+|—|+] °°} +) +)+)+)+)+}—j+ Gad Fnac fmf iI | ml - ‘1O€ES 1%
per ‘d ‘y soua8oree snowy ‘sg f° */—|-+]-+]°°}+)+]+)—|—|+]+\—|-|-|-|—-]+|-|-|-]+}4+ +]-|+/-|-|-} - 9 - ‘oorrirez
gSi -d ‘vaine siyoet ‘sq °° |+|—|—]**|—}-+] F]—-] 4+) 4+) 4+\4+)}— ce eal A “Eye aR be Fl - - ‘Goeee 11
gS1 ‘d ‘nyxaqoy soe “sq f° *|+)—|+]°*|-+]+|+/—|+]+)+}—-|—|— +[-]+]+]+]+/4+]+]+/-|-|-|- > ‘ootee-rzz
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STORRS AGRICULTURAL EXPERIMENT STATION.

BACTERIOLOGICAL INDEX.

Bacillus acidificans,

PAGE

- 158

aerolactis, - 174
butyri rubri, - 168
coli communis, - 186
aerogenes, - 185
disenteriae, - 167
fluorescens minutissimus, - 162
janthinus, - 173
lactarius, - 129
lactis acidi, 114, 135
aerogenes, Vy See
amberis, - 178
arborescens II., - 171
circulans, - 174
citreus, - 166
citronus, - 169
cloacae, - - 180
cochleatus, - 181
Colchesterii, - - 165
contorta, > 155
Cromwellii, - 173
diffusus, - 181
distortus, - 175
erythrogenes, - 117
fragariae, - 167
fluorescens, 161-163, 170
fungiformis, - - 179
gelatinosus, - 176
Harrisonii, - 169
Isignii, - 161
minutus, ~ 160
molecularis, - 160
moruloideus, ° + 163
mycoides, - 172
nebulus, - 165
niger, - 170
nigroferous, - 164
olivaceus, - 160
plicatus, - 177
Pruchii, - = 17h 179
rhizoides, - 171
Robertii, - 182
rubifaciens, - 166
sulcatus, - 166
tenuis, - 176
tetragenes, - 175
vinus, - 178
Zenkeri, - 164

Bacillus mesentericus fuscus,
prodogiosus,
subtilis, -
syncyanus,
violaceus,
vulgaris, -
Bacterium lactis acidi,
album,
arboreus,
Ashtonii,
aureum,
brevis,
Burri, -
catenensis,
chromatum,
citreum,
citronus,
cloacae,
Connii,
erythrogenes,
filiformis,
flocculus,
fluorescens,
Genevum, -
Gorinii,
Isignii,
limburgii, -
liquaerogenes,
lobatum,
luteum,
Marshallii,
Michiganii,
minutissimum,
magnuni,
musci,
myceloideum,
non-acidi,
plicatum,
rubrum,
salmonis,
Synxanthum,
truncatum,
% ubiquitum,
viscosum,
rudensis,
siefige milch,
; visco-furcatum,
Galactococcus versicolor,

BACTERIOLOGICAL INDEX.

Micrococcus auranticus,
cinnabaris,
D (Barthel),
Freudenreichii,
lactis acidi,
albidus,
albus,
arborescens,
aureus,
citreus,
citronus, -
erythrogenes,
flavus,
fluorescens,
gigas,
giganteus,
minutissimus,
rosaceus,
tubidus, -
tugosus, -
varians,
viscosus, -
Proteus vulgaris, -
Pseudomonas coli communis,
- fluorescens,
aurea, -
lactis anana, -
aurea,
aerogenes,

PAGE

109
117
109
124
116
123

203
PAGE
Pseudomonas lactis Estenii, - ISt
Eurotas, = 154
filiformis, - 151
granula, - 159
Middletownii, - 150
mina, - 155
nigra, - = 154
Robertii, - - 156
varians, - 158
viridis, - 152
sapolactica,- - 153
pseudo-tuberculosis, - 152
Sarcina lactis acidi, - 125
albus, - 124
aurantica, - 125
lutea, - 124
Stall-luft bacterium, - 122
Staphalococcus mastitis albus, - 123
pyogenes, - 121
pyogenes albus,- - 123
Streptococcus lacticus, II4, II5, 116
lactis aureus, - - 113
citreus, - - 118
fulvus, - 112
Rogeri,- - - 118
viscosus, - 153
pyogenes, oT,
Tyrothrix, - 173

oh ‘ ie e =

PAMPHLET. BINDERS

“oe This is No.1528
f curried in stock in the’ following sizes :
a4 wDg - THICKNESS; HUGH ‘WIDE THICKNES?
1528 9; inches i webes ¥4,neh iS wz ioches 19 inches 34 iach,
«6, = oS wt “«-
«“ Me: 4 4933 14 att aes ad
S Pee. k, oa
: Other 8s made to order’
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