.BACTERIA LM
te
.3ubitted to
MILE.....
faculty of the
OREGON AGRICULTURAL COLLEGE.
for the degree
o
BACHELOR OF SCIECE
In
DOSTIC sI:LcE
A1D ART.
by
Laura
E.
Ruth
J.
Jaekon.
ließe.
June 1910.
APPROVED:
Redacted for privacy
Redacted for privacy
Dean SchQi of
othei
Säiie
anl Art.
II
ITRODUCTIO1'.
Baoterioloy
is one of
t'ne
sciences, only recentl'
The objects with which it deals are so sLcall
developed.
that they carnot be seen by the naked eye;
and the results
of their activity are so comion, as for instance decay,
souring of milk an
diseae, that they were looked upon
as natural and no cause was ever drearned of.
The first microscope was invented by Hans and Zach-
anus Janson
tu
in 1590;
but bacteria were not discovered un-
sowe time later and then by the use of a simple con-
vex lens.
The discovery was made by a man, now c.11ed the
"father o± microscopy", Anton
Holland,
vho was an
Van Leuwenhock of
.i)elft,
apprentice to a cloth merchant and had
for this purpose learned lens grinding.
Van Leuwenhock's
work was interrupted by tue advent of the bubonic plague
through
urope;
but the application of his discovery to
disease was uAade only by his foltowers.
carried
In 1762 Plenciz
the work along the coLwxion lines of decoiposition
again but the
roress
of the work was again side-tracked
by that farnous discussion which lasted a hundred years fro
L50
to
185), when the theory of 'spontaneous generation'
of life was advanced.
Disproving this theory,
pa1lanzani
showed that by prolonged boiling and excLusion of air, a
meat inl.usion could be kept from putrefying.
In the 18th century the science made a decided
The íiicroscope was iLproved, the c..use of blue milk
-1-
advane.
and
te
connection between bacteria and disease were dis-
covered.
The naine,
until
bacteria,
was
not app1ie
to the or-
botanist, showed that
bacteria were a separate group of plants frein the yeasts
and aniinalcules and ap1ied the Greek naue Ba'1 T por,
meaning, "little stick" to them.
The first classification
of bacteria was made about 1786.
Pasteur proved that the
souring of milk was due to bacteria by the ordinar flrocesses of reproduction and growth. He also proved that
specific organisms caused specific diseases and he did the
greatest wori in interesting the world in this important
ganisins
1617 when Hoffinan, a
Science.
Bonnet, in 1762, originated the theory that bacteria
egs in order to account for irregularities in
sterilizing. Schrdder and Dusch first used cotton 'vool
for filtering air in bactrriological experiments.
might have
Pasteur showed a close connection
the souring of milk, it was ot until
Althouh, about 1860,
between
some
bacteria
and
twenty years ago that theì were found to have a very
close relation to dairying, by M1ler of openhagen, who
divided them into two classes "monas" and "vibrio".
Further o1assifiation as uade by hrenberg, in 18.33, who
with the aid of a better microscope divided them into four
classes, "bacterium", "vibrio", "sprillum' and 'apirochaeta.
Bacteriology is such a broad subject that a life-time
spent in its perusal would fail of an entire knowledge of
the subject, so the authors have chosen a field which is
-
-
certainly one that ought to interest every individual because it pertains to milk, one of the most important foods
of the world.
The advancement in decreasing the death-
rate of infants in the large cities by scientific methods
of handling milk is worthy of attention; and it is the
purpose of this thesis to help in the work of disseinating the knowledge of these methods throughout the country
and especially the dairy districts of Oregon.
The sam-
ples of milk tested were taken from the mixed milk of
dairies and the milk of private cows in and about Corvallis, Oregon.
Those presented here will probably be found
most common to all milk.
BODY OF THESIS.
Bacteria are unicellular plants o± nicroscovic diuensions multiplying
fission and commonly found to be de-
b
void of chiorophyl.
teria
Sorne
be gained from
idea
of'
the ninuteness of bac-
the fact that there íiay easily
be iiore than one billion in a single drop of milk.
average diameter of a transverse section is .001
i
micro
iii.
The
..
or
The forms of bacteria have been corn-
illiaietre.
pared to lead-pencils, billiar-ba1ls and cork-screws.
Bacteria consist of an internal part or proto)las
ed by a cell wall or
less but some
inbrane.
surround-
any bacteria are iuotion-
ove freely about by weans of thread like
flagella and are thus enabled to come in contact with their
Bacteria reproiluce by means of fission but soie
food.
have the power to produce spores.
The spores however are
not a means of reproduction but a means of resisting ad-
verse conditions.
spores require a temperature of
iiany
dry heat for fifteen minutes to destroy them.
perature of from 10°
C.
to 400 C.
is most conducive
A temto
their growth and for most bacteria a temperature of 10
C.
is destructive.
0
Although at the moment of secretion,
milk from a healthy udder is said to be absolutely free
from bacteria, it is a very difficult problem to prove it.
About thirty years ago experiments were made with some
success to prove that wilk drawn with special precautions
into sterilized vessels was practically free from b:cteria.
However,
in recent years,
-4-
even with all our improvements
in aseptic precautions,
it has been
impossible to draw
any quantity of milk which was free from bcteria.
Vlhile
teria.
the milk is in the glands,
it
is free from bac-
This has be n proven by making a bacteriological
examination of the udder directly after the animal has been
slain.
One of the sources of contamination is from the dis-
eased udder.
It has been found that
the tubercie bacil-
lus sometimes locates itself in the mammary glands and
causes infection.
be so slight
:.any times,
however,
that there is no external indication.
milk secreted under these condition
tena.
the lesion may
The
is laden vith bac-
The question is often raised as to whether, from
a cow suffering from a bacterial öiseasr,
not find their way into the milk.
t}e bacteria do
Conn says,
iiacteria
press through the gland only when it is itself the seat of
the
inflammatory process.
Yet a more common source of contamination of the milk
w:ile yet in the 'udder is the entrance of bacteria from
the outside through the teats.
ally would be supposed,
'.hen tnis
Occurs, as natur-
the milk drawn first contains the
greatest number of organisms, while the last will be almost
sterile.
Thus the practice of not using the first fev
jets of milk is justified.
ihe most peculiar occurrence
connected with the bacteria present
the constancy of their numbers,
in
in the
milk-ducts is
though the time of milk-
may differ widely thus, although milk is not drawn
-5-
for several days,
occur
i
souri,
no ba3terial chance, as
the udder.
will
This indicates that there is sozie
check to their growth, iost probably,
erties of the udder fluid.
gericida1 prop-
the
Another circuistance worthy
of notice is thut although the lactic acid bacteria are
norually found in coziparatively
milkin, they
1are numbers shortly after
This
are very seldom found in the ducts.
shows that they are an exterior contamination.
This
brings us to the numerous causes of contaiinations which
take place after the uiik is drawn;
the stable,
the cow,
the iidlker and the uiilking utensils theiselves.
Here, again the cow is an important factor.
the best conditions,
and
ilì
Even under
dust will be present on the cow's body
ay fall into the
in uiovinc about, loose hairs and dust
.
the air or
The likelihood of contamination is much
the cow is not kept in the best condition.
auwented if
If hay is dis-
the air may be
tributed during or just before milking,
filled with myriads of dust particles and with it the
teria which have previously grown upon the hay.
er,
bac-
The milk-
although it is unnecessary, is often a source of con-
taination.
His hands and clothing
and many times of a pathogenic type.
xnay
It
carry bacteria
is usually by
this means and by the water in which the milk vessels are
washed that such a disease as typhoid fever is spread.
Sedgwick and batchelder found on an
obtained in a clean stable,
frorn a
verae, "that milk
well kept cow, by
milking in the usual way into a sterilized bottle, con-
tamed
530 bacteria per cubic centithetre".
"When, how-
ever,
the uAilk man used the ordinary iri1k pall of flaring
form,
seated himself with more or loss disturbance of bed-
ding, and vigorously shook the udder over the rail during
the usual process of milking,
the numbers were very much
higher, on an average 30,500 per cubic centimetre,
mediately after milking."
Sternberg says,
lin-
"the average
of fifteen samples taken from the tables of persons living
in the suburbs of boston was 69,143
per cubic centimetre.
The average of fifty-se!en samples of Boston milk, obtained
directly from the milk wagons and plated at once
2,3b5,500 per cubic centimetre.
T'ae
;as
average of sixteen
sam)les from groceries in the city of Boston was 4,577,000
per cubic centimetre"
some cities,
since milk-inspection has becn adopted,
condemn milk containing over 200,000 bacteria per cubic centimetre.
Great stress must be laid on the sources of contam-
intion and everything should
bilities of contamination.
be
done to reduce the possi-
Still even a more i1portant
coint to be emphasized is the care of the milk after milking.
The lack of good management in this respect is well
illustrated by the enormous increase of bacteria of the
common varieties in sample IX having a count of bl6,00
per cubic ceìtiinetre which was milked and planted in rela-
tively the same time as sample VI having a count of 19,000
per cubic
c
ntimetre.
The former having been left standing
-7-
itchen of the house while the latter had
exposed in the
been carefully handled and kept
cool..
The first step taken to study the bacteria in
ben
after it has
5ilutions of the
brought
to
the laboratory,
ljAilk,
ake
it to
i1k with sterilized water.
A one tenth dilution is made by taking one cubic
centimetre of the milk and adding to it nine cubic centThen for one one-thousandth
imetres of sterile water.
dilution,
is added
one cubic centimetre of the one tenth dilution
to
ninety-nine cubic centiwetres of water.
one-hundreth dilution, one cubic centimetre of
a one
?or
he
milk is added to ninety-nine cubic centimetres of sterile
w at er.
This diluting is done because the milk is so opaque
that it
?.ou)..d
be
plate cultures.
impo.ible
to
see the colonies in the
ihe milk is then plated out.
his
consists in adling one cubic centimetre of one of the
?nown milk dilutions to about ten cubic centimetres of
agar agar, that has been cooled to body temperature,
and after vigorously shaking pouring it into a petrie
dish.
:he agar agar is a nutrient medium used for grow-
ing bacteria.
hese plates are then allov.ed to stand
until the agar solidifies vhen they are
1aced in an
incubator and left for about twenty-four hours at a temp'rature of ninety-eight degrees Fahrenheit.
While in the incubator the organisms multiply rapidly and at the end of
the twenty-four hours it is pos-
sible to count the colonies.
-t
-
A colony is the result of multiplication of one or-
ganism.
Thus the count of the number of colonies rep-
resent
the origir.al number of organisrns in the milk.
A count
of two thousand colonies on a one one-hundredth
dilution plate would have to be multiplied bj one hun
dred and it would then represent the number of organisms
in one cubic centimetre of the milk.
Then by the aid of a sterilized platinum needle the
colonies are segregated and transmitted to the various
media employed.
The kinds of nutrient media used are agar slants,
cause they can be incubated; gelatine,
to show whether
the organism liqueties or not;
to show
the organism develops spores;
potato,
be-
whether
litmus milk, to show whether
the organism produces an acid or alkaline reaction;
plain
milk, to show whether the organism coagulates and re-
dissolves the casein.
Then by the aid of a microscope the morphology of
the organism is examined in a
haning
drop and on a smear
preparat ion.
The motility is shown in a hanging drop.
his is
made by taking with a platinum loop one icopful of liuefled gelatine or some liquid culture, on a cover glass.
he cover glass with its hanging drop is inverted over
the cavity of a hollow slide and
is then examined to
ascertain the motility of the organisms and the character
of their movements.
-9-
To make a smear preparation a straight platinum
needle is used, and the
oranims
the inedia on which they are
are taken from any of
,rowing.
As thin a film as
possible is smeared on the cover glass and allowed to
After it is dry it is passed three
dry spontaneously.
times through a flame, of a Bunsen burner, to fix the
filai
on the cover slip and it is then covered with suit-
able staining solution which is allowe
the organisms are sufficiently, stained.
thorouhly washed
dried.
It
to remove superfluous
is then
to remain until
Then it is
stain and again
mounted with ¿anada Balsam on a
mi-
croscope slide.
The slide is then placed on the stage of the micro-
scope,
a drop o1 cedar
oil being placed on the cover class.
The objective is immersed
in the oil and it is ready for
exam mati on.
The smear preparation is for the purpose of deter-
mining the morphology of the organism.
-1
-
I.
i
':
:,
:..:
',:
I
I
I
,
s?''
s'
-
b
,
,
'.
JI
;
I.
.
,
,
*
j
I
P
'q,
:
:;*i;
;S
/
FIG.
Photograph of i/ic
A.
dilution plate.
ficial,
Showing super-
deep, chromogenic, non-
chrozlioen1c, ani spreading
colonies.
r
I
"\4
,_J"
FIG.
B.
Photograph of agar-
agar plate upon which four hairs
were placed while the agar-asar
was melted.
The photograph shows
the growth o± the organisms which
were upon the hairs at the time
of planting.
150 organisms.
Upon one hair are
This illustrates
the amount of contamination intro-
duced bi one hair falling into
milk.
V
.1-
Painting of the plate
photographed in Fig. B. to show
different colorin6 of colonies.
FIG. C.
%.
n-
'fr
.
:
(
J
;fr ;l
(
ì'
.
q
L
FIG.
D.
2hotoraph of very un-
sanitary methods of treating
and
lii1
bad conditions surrounding the
animals from which tEe milk was
obtaine'L
FIG.
E.
Photograph showing
well entirely too close to the
stable.
eepae of water from
the stable to the well is almost
certain at this iistance and the
milk vessels washed in this water
are sure
to be
contaminated.
n
FIG.
F.
Photobrajh of College
barn where every thing is kept
as sanitary as possible.
__
F
FIG.
G.
Photograph of sanitary
cow-lot with clean, well kept cows.
C'
Owing
A'T
th..tLAj
to a
1
..e
1acing of the milk
in hot agar-agar, the noriia1
number of colonies did not develop and the sample was dise ara e cl.
-11-
SA.PLE II.
Lov.
L:ilked 5:00 A.
10e,
1909.
M.
.1anted 3:30 P. M.
Average count of plates 5:4,O00
per cubic centimetre.
12-
Sorri pIe lu.
m,coccu5
Cotonij I.
F.
Tervido.u5.
y re Z
rrçic rc, bcapeTsc'lci.
OCJQV
r\tH
r
-
/
j
/
Llrn
toto
L
'..i
fltjI
Colony
I,
figure
Micrococus Fervidosus.
I.
::icrococci--non-rnotile-- non-liquefying.
Aar-aar
plate.
Superficial,
raised, smooth, round,
brown colony with darker brown center by trans-
mitted light.
::ilkywhite with heavier white
center by reflected light.
Atar-agar slant.
mooth,vihite,
thin growth along the
line of inoculation with irregular borders some-
what thickened.
Lon-liouefying--arobjc, facultative--an.-
Gelatine.
arobic.
Potato.
Dry, yellow, warty growth.
tato dark grey or violet.
Milk.
Apparently no reaction.
-i
-
Turns the po-
le
C>iLnj ir
s
s
s'
s,
'
e
rcro scopeTieIc
vi
ccjv5ioit
CJ(IC,tjI
t
Lrnusfl¼.
L.
.
Jc
k
a
Il.
BacilLus Ochraceus.
Colony II, Figure II.
thick rod--slightly ictile--liquefying.
Agar-agar plate.
shiny, yellow, raised colony
hound,
with darker center by reflected light an
cloudy
yellow by transmitted light.
Agar slant.
Shiny,
yellow, thick, spreading growth a-
long the line of inoculation.
elatine stab culture.
funnel shape.
aised.
Liquefaction occurs at first in
Forms thick, brown
icroderìna on
top with streaks o± clear and cloudy gelatine.
Potato.
Thick, granular,
orange
rowth.
Does not dis-
color the potato.
Milk.
Coagulates and afterwards reclissolves.
Litmus milk.
Produces an acid.
-14 -
SOIT\Fte ]I.
Lic)ufocer'S.
CcorLjIV
-
r'
fl1vosc.oreF'e d
Oo
V
i'
Tcte
-
-V,
/
-
t-
Oaønt
--
C)Ioftne
íflùk
l..Tim
tflI&
L.t.JocISQ'.
sse
Colony III was the
Colony IV, Figure III.
faciens.
od
lt.
Bacillus Fluorescens Licjue-
shapes--actively motile.
Thin,spreading colony with scalloped
Agar-agar plate.
edges.
as Colony
Green florescence with reflected 1iht.
Almost transparent in transmitted light.
Spreads over the entire surface of
Agar-agar slant.
the slant and the agar-agar gains a green florese ene e
Liquefies in cup shapo,rapidly spreading to
Gelatine.
Liquefies about one-half inch in five
the edges.
Floor of gelatine has brownish precipitate
days.
and white cicroderma floats on top with a cloudy
layer next it.
Potato.
Turns potato light lavender.
Varty yelLow-
18h growth.
Lilk.
Coagulates the thilk and rediseolves.
LItmUB milk.
Lo acid formed.
-15-
Dec.
SALPLE III.
Li1ked 3:00
P.
2lanted 2:00
P.
.
5,
1909.
Dec.
L.
5.
Dec. 6.
Average count of plates, 18,000
per cubic centithetre.
-1
3
-
r'
C'
a.
e
IUICj'J
L15
Cj u
-
I
il
0,
crocar
FYed
Q cor Tjc.e
O,5crt
CjeIcsTine
Totato
L.fmu
i»
£
fl'»(
j,,,,
Colony A, E'igure IV.
iediu
Bacillus Figurans.
long rods--rounded ends---non-motile--lique-
fying.
Agar-agar plate.
superficial,
light.
Agar slant.
Round, shiny,
smooth,
edged colony.
raised and cream colored by reflected
Brown by transmitted lieht.
Iaised,canary yellow growth just along
the iaipfrish.
Gelatine stick culture.
Liquefies in funnel shape
with heavy microderma on top.
Leaves the gelatine
somewhat cloudy.
Potato.
Heavy canary growth which later turns darker
yellow.
itu.us
:il.
illk.
Potato later turns a dirty grey or violet.
son-acid forming.
£IQ5 not coagulate the casein.
-17-
5cn-ri
cocnLj
r
í3cc;(i45 fleblts
.
Tiuve
'
'I'
Cc)CY T'ote
A1
.1
L
i,
2
CY
31oct
-
Tc'ro
¿tJ.ckn
Colony B, Figure
V.
Bacillus
ebilus.
Short rods with rounded ends--non-motile--liquefying.
Agar-agar plate.
Iound, white colony with radiating
lines toward the outside.
Creau by transmitted
light.
Agar slant.
Raised, heavy, white growth along the
line of inoculation.
Potato.
Light brown siiear.
Turns potato nearly the
same color.
Gelatine.
I1ilk.
Liquefies and makes gelatine cloudj.
Does not coagulate the casein.
Litmus milk.
non-acid forming.
-16-
5
CÓ1-
E.
I,- f,
C.
T
cj
u e
'
flic
YósopeF1
Tio-e
Cj eÌTTe
Li
ri
Colony
C,
Figure VI.
Licrococcus
lavus Liquefaciens.
'.:icrococcus ,non-motile---liquefying.
Agar-agar
plate.
LreaLa by
Agar slant.
Oval, yellow,.shiny, raised
colony.
transmitted light.
Smooth, raised, dark
yellow or orange
growth along line or inculat ion.
Gelatine stici
Liquefies
culture.
with heavy
microäera
on top.
Potato.
Slight,
discolor the
iilk.
t)ith,
canary yellow growth.
potato.
Does not coagulate the casein.
Litmus milk.
ìon-acid
forming.
-19-
Does not
C0 Ion D. J3ociii
T%cve
3erfc
L,)
;cIs.
.
'
-s
-s."s
-s
NN
fi
CIcjc
5Iort
ro
co re
Fielci.
Ticre
Lrn..s Çflk
L.Jcck5on
iacillus uperficialis.
tion like it had a teruinal flagella.
Colony D, Figure VII.
hod--slow
Involution forms found on potato. Liquefies.
Vthite, raised colony with thickened
Agar-agar plate.
raised sezrated edges, Vhite with thick grey edge
by transimmitted
light.
slant. V.'hite growth just along the line of inoculation.
Gelatine stick culture.
Iiquofies with heavy white
iaicrodermna on the top and suspended heavy floculi.
Very slight yellow growth just along the
Potato.
line of inoculation. Does not discolor the
Agar
potato.
Milk.
Does not
i.itmnus uailk.
coagulate the casein.
ÂCjd forthing.
-
o-
C&.,
E.
1flcroCLC;t.yc,,,1Icict.s.
2:111:
2
.\
:;
c5ccre
Fd
Colony E, Fiure VItt.
Liierococcus
liquefying.
Agar-agar plate.
of'
1icrococcus Aurantiacus.
staphlo type--non-motile--nonGrows best in incubator.
raised,
oun9,
orange colonies in
bunches.
Agar-agar slant.
Crows just along the iipfrish in
little hard knobs.
t.elatine stab culture.
Potato.
non-liquefying.
Grows well along the iipfrish in little knobs.
Does not discolor.
Li1k.
Does not coagulate.
Litmus milk.
Forms acid.
-21-
SAPLE
Jan.
IV.
1i1ked Jan.
12, 1910.
11 at 5:00 P. M.
Planteò Jan 12 at 3:00
P.
M.
Average count 1650 per cubic
centimetre.
22-
5c:.lc
j
Ccfc-.j I-
Ocjcii-
Ooy
Icjft
fT1u5coides
T'Icte
o
Lt-1
iflk
n-1
L
Bacillus ::uecoiaes.
Short rods with rounded ends--actively thotile.
IonSpores are formed at the ends of the rods.
liquefying.
White, spreading colony, grows in fancy
Agar-agar plate.
shapes like ferns.
'Vhite, spreading colony vith fern like proAgar slant.
Colony
I, Figure
IX.
ject ions.
Dirty brown with tiny raised creases in a),.l
directions, spreading and turns the potato a dirty
white.
ion-liquefying.
Gelatine stab culture.
Does not coagulate the casein.
Liilk.
Lon-acid fori.ina.
LitnLus uìilk.
Potato.
-23 -
SPE
V.
ìi1ked
6:0') A.
Jan.
26,
1910.
M.
Plantod 3:00 P. M.
Average count 54,000 per cubic
Centithetre.
-24-
5cripie.
C oIrt'j O. J3cciIIu
I)W'
QYO (Oc'.5
..
t.
I
1flicrtd.
T'çcte
¿
Ï
ZIc,rl
j-
(3elottfl
L
f
n
i
i
k
i
muS rn1 1k
L.t.,Jek Son.
Bacillus Aerolactis.
Colony A, Figure X.
(Conn)
Ioòs small with rounded ends--non-motile--prolific
spore producer--liquefying.
Agar-agar plate.
Agar slant.
Thite,
spreading, wrinkled colony.
hite, rather streaked growth along the
iwpfrish.
Gelatine stab culture.
Potato.
Milk.
Liquefies and forms a floculi.
Dirty yellowish brow:.
Coagulates the casein.
Litmus mil1.
on-aciä forming.
25-
Discolors the potato.
5csrr
C
3;u
Ocac'i
t
.ive
t'
/
C(c)cr
-iote
V
L14
s
4-
L\_
Oov
5I,,t
ThuR
LEicucI.uscuu,
I
colony B
Colony
saiie
,
as
C.
acillus Acidi Lactici.
Figure XI.
arge rod with rounded ends.
Foruis spores in
Brownian
the center of the rod.
AOt1Ofl.
iOfl-
li u e fyi ng.
Agar-agar plate.
Vhite, aluiost transparent, deep
growing colony.
Agar slant.
Transparent white colony gro'ing only
along the impfrish.
e1atine stab culture.
Potato.
Non-liquefying.
Spreading, white growth--raised, wrinled and
tenacious and turns the potato yellowish violet.
iblilk.
Coagulates and redissolves the casein.
Litmus milk.
Acid
forming.
-26-
SAL1PLE VI.
Jan.
Lilked 6:00
Planted 3:00
P.
P.
M.
M.
3,
1910.
Jan.
3.
Jan.4.
Average count 19,0D per cubic
centimetre.
e)
L.
LLjJ7
n
I
O
A
LtLL Ql
¡J_
I
/
L
L
'
/
r-'
11z
Çr,z,,
°-1Lu
jaI1tuG-
oIc)
.r-n
Colony W, Figure XII.
Micrococcus Aurantiacus.
Very athall Lcicrococ3us--facultative anaerobic,
non-uotile--non liquefying.
Agar-agar plate.
Agar slant.
Round, greenish
'ellow colony.
Very pretty greenish yellow growth along
the line of inoculation.
Gelatine stab culture.
1on-liquefying.
Good growth
along irnpfrish.
Potato.
Shiny yellow growth just along the line of
inoculation.
iik.
1itus
Does not disco.or the potato.
Coagulates and redissolves the casein.
iiilk.
Acid forzidng.
-?
3
-
5rnf1e
ce-'
i
y
,-
E.
mcrocc.. Ccdt
xiii
/
:
//
flhicvOzcÖ?iieic.
L)
-
L,iu
Ik
Colony V, Figure
XIII.
::icrococcus Candicans.
;iicrococci, facultative anaerobio ,non-inotile--lique-
fying.
Agar-agar plate.
cream colored diamond
shaped
Crearnish yellow with heavy
edges.
Deep
colony.
Agar-agar slant.
elatine stab culture.
Liquefies in cup shape and dries
out as fast as it liquefies.
Potato.
Milk.
Verd slight growth.
Does not discolor.
Does not coagulate the casein.
Litmus milk.
1on-acid forming.
'-'n
fl
Z
:x:Ir.
4.
íflicocorTlId
Ci
C)
or
A
H,
'i
090 r5/ort
-v,1-ct
G
1k
LTi,i..5 flilk
LLJack,or
Colony
Z,
Figure XIV.
Saccharoiyces
ossceus.
Large round bodies about three idcrons long.
àppeai
granular inside.
Agar-agar plate.
ound,
pink colony with dark pink
center.
Agar-agar slant.
Heavy, shiny, sali.on-pink growth along
the line of inoculation.
(e1atine stab culture.
ion-liquefying.
Button like
growth at the top and slight growth along the line
or inoculation.
Potato.
Heavy
paint.
:11k.
pini:
growth In two days.
Looks like wet
Iot shiny.
Does not coagulate the casein.
Litmus milk.
ion-acid forming.
This 'east makes a bread with a very delicate crust.
-30-
SAMPLE VII.
iarch 11, 1910.
11ked 7:00 A. M.
Planted 2:30
P.
M.
iarch 11.
:.:arch
11.
Average count 600 per cubic
centimetre.
:ost1y BaciLlus Luscoides.
new colonies.
-:-
Lo
March
SALPrJE VIII.
Li),kecl
6:00 1. L.
Planted 3:00
P.
M.
6,
1910.
íarch 5.
March
6.
Average count 45,000 per cubic
centimetre.
-Z2-
VII
CcloI) T
i
I45 C'youlc,1-5
iicire
I'
-
7c
í11yo
t
milk.
Colony
I,
Figure
V.
Bacillus Circulans.
Small rod about one and one-half microns long.
Prolific spore producer.
Actively zotile.
Liquefying.
Agar-agar plate.
iaised, white, shiny superficial
colony with Lan
filauents.
Brown by trensiitted
light.
Agar slant.
iaised,flesh colored, slightly wrinkled
growth just along the
Gelatine.
ixzipfrish.
Liquefies very slowly in cup shape and dries
out as fast as it liquefies.
Potato.
i'aised,
wrinkled, spreading
the potato.
Milk.
Does
Litmus milk.
nt
coaulate the casein.
ìon-acid forming.
rowth and discolors
SALPLE IX.
klLilkeã
April 11, 1910.
5:CO P. M.
Planted 3:00
P.
April
i..
9.
April ii.
Average count 516,300 per cubic
centimetre.
o
new cdonies.
SAIPLE X.
April 11, 1910.
Milked 6:00
A.
Planted 3:00
Average
count
centitLetre.
I.
P.
M.
April 11.
April 11.
650 per cubic
The
count vas so
low that a preservative was
pee ted.
-35-
sus-
SAii?LE XI.
i1ked 6:00
Lpril 15, 1910.
A.
April 15.
M.
Planted 3:00 P. M.
April 15.
Same milk as sample X.
ed
Test-
for preservative but none
was found.
io colonies on the
1/loo or the 1/1000 dilution
plate.
15,600
iiate.
Irregularit' not
ed for.
on i/iC dilution
ecount-.
1
tTT
9fl
0
UI
'2
OPT
6061
TTH
COO'O9'T ETUOTOO
tIflO3
T
AO
0
-ç-
o
rncvcocu
u
CH
r
C)oCo ?FIe4.
C.orT'
L-'
IIC.
__)Q
-
--- )-leç
's'
Description of Figure
Plate II.
Coloni II;
I.
iicr000ccus Albue.
By tranaadtted light on agar plate.
Small,
raised, smooth, homogenous mass.
oval,
By reflected light.
superficial,
Light brown.
Luilky white.
Thin, white film showing inany cross mark-
Agar slant.
i ng s.
Dry, yellow,
Potato.
varty growth.
Growth only along line of inoculation.
ender.
Gelatine stab.
1on-liquefying aerobic facultative
anaerobic.
Litmus milk.
Milk.
Form.
o
Turns potato lay-
io action.
action.
Small micro-cocci.
-39-
LIí3
SnW 4t
)jt'UJ
II
40kLAQ(zD
'I
r
3AI2a
s.0
I
Su
çA,cr
t
-1[cLTLEIu
OQ7
Description of Figure
Colony III, Plate II.
II.
Bacillus Flurescens Liquefaciens.
By reflected light on agar plate.
colony.
Agar slant.
Greenish white, deep
Loss-like growth with many filaments.
Vhite, spreading growth.
Agar assumes
green florescence.
Potato.
Yellow, warty growth.
Turns potato lavender.
Liquefies gelatine precipitate on floor
Gelatine stab.
of gelatine.
Litmus milk.
Coagulates and redissolves casein.
Acid
react ior.
Milk.
Coagulates
Form.
Short,
ad
redisolves casein.
thick rods usually in pairs and slightly
motile.
-40--
-
(cQy5 cç+
Cyotu-te
!CQfl5
LiTflu.sfl)i 1k
fl1
1k
Description of Figure III.
Colony IV, Plate II.
Bacillus Dchracens.
By reflected lieht on agar plate.
Small,
round,
smooth,
yellowish i;hite, raised, shins colony having darker
center.
By transmitted light.
Dee; brovn center.
Outer part
lighter brovn.
Agar slant.
Smooth,
shiny; yellowish growth.
Growth
only along line o± inoculation.
Potato.
Orange colored granular growth.
Growth only
along line of inocul3.tion.
Gelatine stab.
In gelatine stab cultures liquefaction
occurs in funnel form.
Forms a brown uicroderma
on floor of gelatine and a brown 9eposit at point
of funnel.
Cloudy on floor of gelatine then clear
streak then cloudy strea:.
Litmus ilk.
Coagulates and redissolves casein.
reaction.
:.i1k.
Form.
Coagulates and redissolves.
hort slender bacillus.
-41-
Motile.
Acid
SAMPLE III.
I1i1ked
3:00
Plates
ae
From 1ing's
P.
2:00
Count 18,000.
to pick up
M.
P.
iry.
Dec.
5,
£&.
Dec.
1909.
6,
It was impossible
anzi
colonies as there
was a white filmy colony over the
entire plate.
-
o
.
¿
-
SAIOELE IV.
i1ke
A.
.'roni
I.
Dec.
.1ates made P. I.
-'-
O.
A.
15,
Dee.
.
Larin.
19O.
15, 1909.
Scirnp)eI.
CoI.-Ip
1L,ye
].
Ifl.crocc,reTfcj.
r
CIorlrft
Toi-ot(,
t.
li-n
'-'S
rTl
k
mn
Description of Figure IV.
Colony
I,
Bacillus Fluivis.
2late III.
By reflected light on agar plate.
Light brown.
By transmitted light.
slant.
Raised, smooth, shiny, lemon yellow growth.
only
Growth
along
lino of inoculation.
Lemon yellow, shiny colony.
Fotato.
yello'i.,
colony.
raised,, superficial
Agar
Small,oval, shiny,
line of inoculation.
rowth
only along
Vrinkled with deep fissures.
Turns çotato dirty v1hite.
Liquefies gelatine.
Gelatine stLb.
tive anerobjc.
ir.
Form.
Liuefaction very slow.
It
occurs
cup shaped form.
Litmus milk.
iD-.
Aerobic faculta-
i:o
o
action.
action.
Short rods.
Transverse divisions.
-44-
ion-motile.
"H
r
I'UJ
1LIW9flLU4t
A
7LA
.z"__-
oA'(,(j
rj
T_j
Ji1iuoi-,
Description of
igure
Colony II, Filate III.
V.
::icrococcus Lactis Aureus.
By reflected light on agar plate.
round,
Small, raised,
shiny, flesh colored, superficial colony.
By transmitted light.
Agar slant.
growth.
potato.
Smooth,
.aight brown.
raised,
shiny,
orange colored
Growth only along line of inoculation.
Shiny, orange colored growth.
dirty grey.
Gelatine stab.
Growth only alon
Turns potato
line cf inoculation.
Loes not liquefy gelatine.
microderma on floor
Litmus milk.
(Conn)
Orane
Acid reaction.
o1
yellow
gelatine.
microderiia on floor or udlk.
Action very slow.
Coagulates
milk.
Mill.
Sliaht yellow growth around tube at upper surface of milk.
'orm.
Coagulates milk.
Ledium small mlcTo-c000i.
-45-
Sörn pIe!E.
C0j091 ?JI flhICYCøCCLJ5
Tcre t
L-oc
¡
Voy
s
t
O
p.
f'
Oqy.$)onf
L1rru
Vflilk
1flk
T.JJ1ess.
Descrition of Figure
Colony
I,
VI.
.iicrococcus Lactis Varians.
Plate II.
By reflected tight on agar plate.
mall, raised, shiny
colony, with serrated edges.
By transmitted 1ig1t.
Agar slant.
Light brown.
Light ochrans yellow growth.
aised an
wrinkled with deep fissures.
Dull flesh colored granular growth.
2otato.
only along line
01'
inoculation.
rowth
urns potato
dirty grey.
Gelatine stab.
Liquefies gelatine in cup shaped form.
Aerobic facultative anaerobic.
Litmus
m11ì:.
milk.
illk.
Yellow inicroderma formed on floor of
Coagulates milk.
Coagulates milk.
Yellow microderma formed on
floor of milk.
Form.
Very small
micro-cocci.
-46-
fl iCi'OCOCCLJ FYV1cto15
TuveYJL
CoIor.jjrpJj
C)
5ict
T
to
te
rrtIk
TJ.)1esj.
Description of Figure VII.
Co1oni II, I?late II.
ìicrococcus ?ervidosus.
Small,
By reflected light on agar plate.
smooth,
shiny,
superficial, white colony.
By transwitted light.
Light brown.
Smooth, shiny, white growth.
Agar slant.
Growth only
along line of inoculation.
hite, shiny growth.
Potato.
Turns potato dirty white.
Growth very scant.
Celatine.
Does not liquefy gelatine.
scant.
Litmus milk.
1'o
action.
LlIilk.
10 action.
Form.
Diplo-òocci--non-ciotile.
-47-
rowth very
5rrpIei
C tioThy,., D;cIt-.
TCjUYe
ILLL.
r
rn
5iont
rie
LI-
c
ro
c
pe'T; e'
-
r-11k
Description of Figure VIII.
Colony III,
'late
II.
Ulaothrix Dichatoma.
By reflected light on agar plate.
ficial, shiny colony.
Smooth, white,
super-
Opacjue with dull white
center.
By transmitted light.
Asar slant.
bight brown
vith white center.
aised, white, warty growth.
iives off
odor of mould or decaying wood.
Same growth as on agar slant.
Potato.
Gelatine.
Liquefies gelatine in cup shaped form.
Aerobic facultative anaerobic.
LitLLus milk.
Casein is coa-ulated and redissolved.
Acid reaction.
milk.
Light yellow growth around tube on surface of
milk.
Form.
Coagulates an9 redissolves milk.
Long thread -like filaments having group of
spores at the ends.
The filaments are branching.
From Irvine's, Private cow.
SAMPLE V.
;.:ilked
A.
.
Jan 14, 1910.
Platee made 2:30
Count 19,200.
-49-
P. M.
Jan.
14.
rnI te
cOIon)]rPJn
rÑure
li
J3ciIusUbu1tL-
X
Çllico
opeFec.
Qc1y PIote.
JI
F
-
Cic)cr5IOflt
fl-1ik\
Cjlcitine
Øto1C)
1.J
t-ls
Description or Figure
late III
Colony II,
X.
Bacillus Ubiquitous.
By reflected light on agar plate.
Deep white, spread-
ing, zioss-like growth, with many filaments.
Clear, white, soread1n
Agar slant.
growth, with many
filaiients.
lear, yellowish white, wrin1led, spreading
Potato.
Turns potato grey.
growth.
Gelatino.
white,
scanty growth.
)oes not liquefy
gelatine.
Litmus milk.
Coagulates and recliscolves casein.
Milk.
Coagulates and redissolves casein.
Form.
3hort thick rods.
Ion-uìotile.
-50-
rr
re
rj1tpjiZ
i-i
re iT
I
/
Field
-
I
-J
Lírnu
rn
k
m1tk.
-WJ
1le35
Description of "igure XI.
Colony III,
late ILl.
acillus iyocanus.
3y rcflected lig1t on agar plate.
round,
Greenish white,
raise1 colony.
By transuîtted light.
ight brown,
dee;er brown center.
:ased, shiny, greenish white, spreading
Agar slant.
growth.
Gives agar green florescence.
Brownish white, granular growth.
Potato.
along line of inoculation.
Gelatine.
V.hite
iquefies gelatine.
Turns potato grey.
Action very slow.
öeposit or floor or gelatine.
along line of inoculation.
Litthus ¡xilk.
Dissolves casein.
L:ilk.
Dissolves casein.
form.
Long rods.
otile.
-51-
Growth only
canty growth
CoIeflj.TJJZC
.b
(/L5
I,
,
l
Corple
2
-
5Icrt
C)eIttr
Votot
jJ reS.
Description
oÍ'
:igure XII.
Colony IV, Plate lit.
l3acillus Cancliccus.
By reflected lignt on agar pla:e.
raised,
shiny clony.
By transmitted light.
Agar slant.
alon
¿mocth, white, round,
Light brown.
hite, raised, shiny growth.
(rowth only
line o± inoculation.
Dirty brownish white, granular grow±.
Potato.
only along line of inoculation.
elatine.
Does not liquefy gelatine.
Growth
Turns potato grey.
Growth scanty
along line or inoculation.
Litmus mili,
Coagulates mi1I.
Light brown deposit
around tube.
::i1k.
orra.
coagulates
miii:.
Short thick rods.
i.on-,otile.
organ's private cow.
VI.
::i1ked A.
Plates
Te
Feb.
7,
ado 2:30 P.
/i.
.
1910.
Feb.
plates made were not goon.
7.
SAI1FLE VII.
Li1ked
P.
Froza Szxiith's
Larch
M.
Plates maae
P.
Count 45,000.
-54-
i.
6,
private 30W.
1910.
March
7,
1910.
-
._cnieyjj_
C&orij]JL PI
JJc,c;j/.,3 Li-ii k,arisC!c ,ds
Loctici
flre zur
i
I
/
lorie
LnrzIk
R-J t-ies.
Description of Figure XIII.
Colony III, Plate
Bacillus Liznbatus Acidi Lactici.
I.
Clear, white, round,
By reflected light on agar plate.
deep colony.
By transmitted light.
Agar slant.
Light
Light brown.
ïel1ow,shiny,
raised growth.
Brownish yellow, moist, shiny, slightly
Potato.
Ing growth.
(e1atine.
Turns potato grey.
Does not liquefy gelatine.
Scant Growth
along line of inoculation.
.itus
milk.
sjread-
Acid reaction.
Milk.
Dissolves casein.
Form.
Short bacillus.
Dissolves casein.
ion.-motile.
-55--
¶3orrçtJ
r1icrococ'S°'
/,-
1/
Ji
JI
crccc,e
o-PIcrte
\
c
inilk
B.L1E5S.
-
Description of Figure XIV.
Colony
Plate III.
I,
icr000ccus Auranticus.
By reflected light on agar plate.
iiaised,
shiny, light
yellow, diauond shaped colony.
By transthitted li;ht.
.4ight
brown.
Orange yellow, with lichter yello.w eages,
Agar slant.
smooth, shiny colony.
ioist,
Potato.
light brown, pink tint, shiny Lrowth.
Growth only along line of inoculation.
Does not liquefy gelatine.
Gelatine.
along line of inoculation.
Litmus milk.
iilk.
orm.
o
3lightly acid.
action.
iicro-cocci.
-56-
Scant growth
1
-S orn
Coso-)
moeocc
11
Coricent%IcL1.
liC)UY
mIcroscope
clocfl,
PIc,fE
II;'
ne
LThII
R.J. 14e$5
Description
Colony IL, Plate
iy
oÍ'
Figure
V.
icrocoocus Concentricus.
III..
reflected light on agar plate.
white,
Agar slant.
Light brown.
Thin film, white, spreading growth.
Dirty white, raised, granular growth.
only along line of inoculation.
Gelatine.
Does not iiiuefy gelatine.
scant along line of in;'culation.
Litmus milk.
Llilk.
Form.
oval, raise3,
Shiny colony.
By transrnitted light.
Potato.
small,
o
Lo action.
action.
Micrococci
sinLly or in paire.
-57-
rov,'th
Turns potato grey.
Growth very
Smtle1.
CIor-1)usVffr rncecs Cat reus.
FIc)uve
i
flmc'occ,ye Fi&td.
CI 90 VP 0Te.
Lrrn
IpIk
tflIk
Description of Figure
iicrooccus
Colony ILL, 2late III.
itrens.
Snal1,
by reflected li,ht on agur plate.
oval, raised,
shiny, yellow colony.
By transnitted light.
Light brown.
Pinkish yellow, shiny growth.
Agar slant.
trovth
very scant.
Orange colored growth, very slight along line
Potato.
of inoculation.
Gelatine.
Does not li:uefy gelatine.
Slight white
growth along line of inoculation.
Litzus
¿iilk.
io action.
.Iilk.
.o action.
Foru.
Cocci
varied in size.
threes or in mosses.
-56-
$ingl,
in pairs,
in