Bacterial flora of the alimentary tract of Grylloblatta campodeiformis campodeiformis Walker
by Albert L Burroughs
A THESIS Submitted to the Graduate Committee in partial fulfillment of the requirements for the
degree of master of Science in Bacteriology at Montana State College
Montana State University
© Copyright by Albert L Burroughs (1941)
Abstract:
A study has been made of such bacteria as may be found in the alimentary tract of Grylloblatta
campodeiformis campodeiformis Walker.
This insect is a cold-loving creature which lives at temperatures near freezing. With due precautions to
avoid intruders, cultures were made from the alimentary tract by plating in dextrose agar. Incubations
were made aerobically and anaerobically at 4°C, 18°C and 28°C. No growth was obtained
anaerobically. Colonies from the aerobic plates were picked and streaked on dextrose agar. The
optimum temperature for all but one culture was found to be room temperature (25°C). This one culture
grew best at 17°C to 22°C.
Morphological studies of each of the cultures was made and inoculations were made into various media
in an attempt to classify the strains of bacteria.
Fifty strains of organisms obtained from two specimens were included in this study. Sixteen (32 per
cent) were found to be gram-positive cocci of the genus Micrococcus, 16 (32 per cent)were
gram-positive rods of the genus Bacterium, and 18 (36 per cent) were gram-positive spore-formers of
the genus Bacillus.
Five of the genus Micrococcus were classified to speoies. Three of these proved to be strains of
Micrococcus epidermidis (Kligler) Hucker, one was variant of Micrococcus subflavus Bumm, and one
a variant of Micrococcus freudenreichii Guilleteau. None of the genus Bacterium could be classified to
species. Of the genus Bacillus, two were members of the Aerobacillus Group. Three strains were of the
Bacillus adhaerens Group, one of which was possibly a variant of Bacillus panis Migula. One strain
was a member of the Bacillus circulans Group and was possibly a variant of Bacillus serusitidus La
Coste. One was a member of the Miscellaneous Group. The other eleven organisms were members of
the Bacillus subtilis Group. BACTERIAL FLORA OF THS ALIMENTARY TRACT OF GRYLLOBLATTA
CA PODBIF',R IIS CA iPODEIFOR T S WALKER
by
ALBERT L . BURROUGHS
A THESIS
S u b n itte d to th o Graduate C oianittee
in
a r t i a l f u l f i l l n e n t o f th e requirem enta
f o r th e d ecre e o f
a s t e r o f S c ie n c e in
'a c te r io lo g y
at
Itontana S ta te C o lle g e
Approved:
In Charge o f
a jo r Work
Jh aim an , Examining Committee
a n , 'rraduate CoTi) lt t e e "
Boze a n , Montana
June, 1941
N
3 7 ^
2
TAULE OF CO:-. TB: TS
A b s t r a c t ....................................................................... ..
In tro d u c tio
lx s r i o n t a l
re s u lts
.................................................................... ....
a t 1 o d s .............................................. ..
................................................ ..
Pa^e
S
4
9
12
D i s c u s s i o n ................................. ................................ ..
40
Sum a r y
45
* ................
L i t e r a t u r e C ite d
..........................
Ii
Sr=*
CD
C3
60105
46
-3
ABS TRACT
A stud y has been made o f such b a c t e r ia a s may be found i n th e
a lim en ta ry t r a c t o f G r y llo b la t t a oam podeiform is cam podeiform is W alker.
T his in s e c t i s a c o ld - lo v in g c r e a tu r e which l i v e s a t tem p eratu res near
f r e e z in g . With due p r e c a u tio n s to a v o id I n tr u d e r s , c u lt u r e s w ere made
from th e a lim en ta ry t r a c t by p la t in g in d ex tro se a g a r . In cu b a tio n s were
made a e r o b ic a lly and a n a e r o b ic a lly a t 4°G, IS0C and 28°C. No growth
was o b ta in ed a n a e r o b ic a lly . C o lo n ies from th e a ero b ic p la t e s were p ick ed
and strea k ed on d e x tr o se a g a r . The optimum tem perature f o r a l l but one
c u ltu r e was found to be room tem perature (2 6 °C ). This one c u lt u r e grew
b e s t a t 17°C t o 22°C.
M orp h ological s t u d ie s o f each o f th e c u ltu r e s was made and
in o c u la t io n s were made i n t o v a rio u s m edia in an attem pt to c l a s s i f y th e
s t r a in s o f b a c t e r ia .
F i f t y s t r a in s o f organism s o b ta in ed from two specim ens were in clu d ed
in t h i s s tu d y . S ix te e n (32 per c e n t) w ere found to be g r a m -p o s itiv e c o c c i
o f th e genus M icro co ccu s, 16 (32 per cen t)w ere g r a m -p o s itiv e rods o f th e
genus Bacterium , and 18 (SG per c e n t) were g r a m -p o s itiv e sp o re-fo rm ers o f th e
genus Iiaol I IusT
F iv e o f th e genus M icrococcus were c l a s s i f i e d to s p e c i e s . Three o f
th e s e proved to be s t r a in s o f Hio ro o o co u s ep id erm id is ( K lig le r ) Huoker,
one was v a r ia n t o f M icrococcus SuljftIavus Bumm, and one a v a r ia n t o f
M icrococcus fr e u d e n r e io h ii G u ille b e a u . None o f th e genus Bacterium co u ld
be c l a s s i f i e d to s p e c i e s . Of the genus B a c i llu s , two were members o f th e
A e r o b a o illu s Group. Three s t r a in s were o f the Baoi l l u s adha eren s Group,
one o f w hich was p o s s ib ly a v a r ia n t o f B a c illu s p anis M ig u la . One s t r a in
was a member o f th e B a c illu s o ir o u la n s Group and was p o s s ib ly a v a r ia n t o f
B a c illu s s e r u s it id u s La C o ste . One was a member o f th e M isc e lla n e o u s Group.
The o th er e le v e n organism s were members o f th e B a c illu s s u b t i l i s Group.
-4 „
BACTERIAL FLORA OF 'HE ALIMEHIAHY TRACT OF GRYLLOBLATTA CAMPODEIFORMIS
CA.MPODBIFORf.il S UALKER
INTRODUCTION
This in v e s t ig a t i o n was undertaken as an attem p t to i s o l a t e and stud y
th e b a c t e r ia l f lo r a o f th e a lim en ta ry t r a c t o f G r y llo b la tta cam podeiform is
oam p od elforais Y alker.
This i n s e c t was d isc o v e r e d i n th e Canadian R ookies
i n 1913 by Dr. E. M. t a lk e r .
th e i n s e c t .
Crampton (1 9 1 5 ) has th e f o llo w in g to say o f
ltG r y llo b la tta cam podeiform is com bines w ith in i t s e l f c h a r a c te r s
common to a number o f ’ O rthopteroid* i n s e c t s .
Indeed in many r e s p e c t s i t
may be c o n sid e r e d a s a v e r i t a b l e li v i n g f o s s i l , and from th e p o in t o f view
o f th e stu d y o f i n s e c t phylogeny i t i s one o f th e m ost im p ortan t o f r e c e n t
p to ry g o ta n fo r m s .”
Gurney (1 9 3 7 ) s t a t e s th a t the d is t r ib u t io n a l data r eg a r d in g G r y llo b la tta
a re s t i l l v e r y fra g m en ta ry .
The ev id e n c e p o in ts to th e p resen ce o f many
w id ely s e p a r a te d , more or l e s s i s o l a t e d , u n it s o f p o p u la tio n .
The th ree
forms in th e p a c i f i c c o a s t s t a t e s a re d i s t i n c t and th e n a tu r a l b a r r ie r s c l e a r .
G r y llo b la tta s o u lle n l occu rs in Oregon.
G r y llo b la tta oampo d elfo rm !s
p o o ld e n t a ils o ccu rs on Mt. Baker, W ashington.
G r y llo b la tta b arb er! occu rs
i n th e S ie r r a Nevada M ountains i n Plumas County, C a lif o r n ia , and i s
sep a ra ted from i t s n e a r e s t Cascade r e l a t i v e by v a lle y s i n th e w atershed o f
th e Klamath R iv e r .
G r y llo b la t t a cam podeiform is cam podeiform is occu rs a t
B a n ff, A lb e r ta and G a lla t in County Montana.
G r y llo b la t t ld a e , G a llo is ia n a occu rs i n Japan.
Another genus o f the
This genus i s rep re se n ted by
two su b gen era, G a llo is ia n a and I s h ia n a , n lp p o n en sis b ein g th e on ly s p e o ie s
o f G a llio s ia n a and r e p r e se n te d by one a d u lt m ale and two nymphs.
The
-6 subgenus I sh ia n a i s r e p r e se n te d by one s p e c ie s , n o t a b i l i s .
A s in g le male
nymph has been tak en o f th e l a t t e r .
M ills and P ep p er, (1 9 3 7 ) found th a t th e tem perature m ost d e s ir e d by
th e s e i n s e c t s was S .7 °C j t h a t s t a r t in g from 3 .7 °C , upon c o o lin g th e r e was
a r e d u c tio n in a c t i v i t y a t OeI 0G and t h a t t h i s le t h a r g ic c o n d itio n co n tin u ed
to - 5 .6 ° C .
At t h i s p o in t th e r e was in c r e a s e d a c t i v i t y f o r a sh o r t tim e ,
p r o s tr a tio n en su in g a t - 6 .2 ° C .
s t a t e b e fo r e p r o s t r a t io n .
A pparently th e in s e c t went in t o no dormant
In t r a v e l lin g up th e tem perature s c a le from
3 .7 °C , th e f i r s t change n o ted was an in c r e a s e in a c t i v i t y a t 1 6 .5 °C .
There
was p a r t i a l p a r a ly s is o f th e le g s a t 2 4 .9 °C , q u ie sc e n se a t 26.4°C and f i n a l
h e a t p r o s tr a tio n a t 2 7 .8 °C .
This i n t e r e s t i n g work on th e tem perature
r e la t io n s h ip s o f t h i s i n s e c t c l e a r ly dem on strates th a t a tem perature which
i s com p atib le f o r m ost i n s e c t s w i l l produce death in th e g r y l l o b l a t t i d .
I t a ls o shows th e g r y l l o b l a t t i d i s v ery a c t iv e a t tem p eratu res a t which
m ost i n s e c t s w ould go in t o dormancy.
A c tiv e g r y l l o b l a t t i d s a re o fte n
c o l l e c t e d from b eneath o b j e c t s which a re fr o z e n to tiie e a r t h .
I n d ic a tio n s from W alker’ s la b o r a to r y in Toronto a re t h a t th e l i f e
c y c le o f th e i n s e c t ta k es about 10 y e a r s .
I t i s unusual i n i t s developm ent
i n th a t th e r e i s a p r e - a d u lt s t a g e , th e su b-im ago.
This i s v e r y u n u su al,
th e Epheme r id a e or M a y -flie s b ein g th e o n ly o th e r in s e c t s w ith such a s t a g e .
During th e sub-im ago th e i n s e c t i s w h ite , l i v e s in th e ground and does n o t
appear a t th e s u r f a c e .
c o lo r .
A fte r th e l a s t m oult i t assumes a honey y e llo w
This in fo rm a tio n has n o t been p u b lish e d to d ate b u t was brought
o u t in a c o n v e r s a tio n betw een Dr. H. B. M ills and D r. E. M. W alker.
An i n s e c t which r e q u ir e s such a low tem perature s c a le m ight p o s s ib ly
-6 be th e so u rce o f a p s y o h r o p h ilio b a c t e r i a l f l o r a .
With t h i s in mind th e
i n v e s t ig a t i o n o f th e f l o r a o f th e a lim en ta ry t r a c t was begun.
L ite r a tu r e on th e s u b je c t o f i n s e c t - b a c t e r i a l r e la t io n s h ip s i s
s u r p r is in g ly li m it e d .
I t would be supposed t h a t , from th e p o s s i b i l i t i e s
o f b i o l o g i c a l c o n tr o l o f i n s e c t s , more in v e s t ig a t i o n s in t h i s f i e l d would
have been made $ a ls o t h a t w ith regard to th e im portance o f i n s e c t s in th e
tr a n s m is sio n o f d is e a s e co m p a ra tiv ely few s p e c ie s have been examined as to
th e ir b a c te r ia l f lo r a .
S tein h a u s (1940) has r e c e n t ly su rveyed th e f i e l d
and p r e s e n ts th e b e s t co m p ila tio n o f p rev io u s work done on t h i s su b je c t
w ith regard to a l l p h ases o f th e b io lo g ic r e la t io n s h ip s e x i s t i n g between
i n s e c t s and th e b a c t e r ia th ey h a rb o r.
Leydig (1 8 6 7 ) n o ted th e p resen ce o f organism s i n th e o a e o a l appendages
o f a p en tatom id .
Forbes i n 1882 d isc o v e r e d numbers o f b a c t e r ia i n th e
oaeoae o f c e r t a in H e te ro p tera .
Glasgow (1 9 1 4 ) w h ile w orking on th e f l o r a o f th e g a s t r ic caeoae
d evelop ed an a s e p t ic method fo r rem oving th e oaeoae as f o l l o w s .
"The
in s e c t i s f i r s t l i g h t l y ch loroform ed to p rev en t s t r u g g lin g , th e w ings are
c lip p e d o f f near th e b a se and th e w hole body m o isten ed w ith a lc o h o l to remove
th e f ilm o f a i r and a llo w th e p e n e tr a tio n o f th e b ic h lo r id e s o lu t io n which
was u s u a lly u sed i n th e p ro p o rtio n o f 1 -6 0 0 .
Ihe m ercuric c h lo r id e s o lu t io n
i s b e s t a p p lie d w ith a b i t o f ab sorb en t c o tto n h e ld i n a p a ir o f o ld f o r c e p s .
In t h i s way th e e n t ir e body o f th e in s e o t can be th o ro u g h ly scrubbed w ith
th e d i s i n f e c t a n t , so t h a t any f o l d s , suoh as th o se betw een the body
segm en ts, w i l l c e r t a in ly be m o isten ed .
A fte r th e b ic h lo r id e s o lu t io n has
co m p lete ly d r ie d , which may be v ery w e ll h a sten ed ty p a s s in g th e in s e c t
7baok and fo r t h b e fo r e a Bunsen fla m e, th e f l a t edges o f th e abdomen are
o lip p e d o f f , from n ear th e p o s t e r io r end up to th e th o r a x , w ith a
f i n e s c i s s o r s which have been p r e v io u s ly fla m ed .
a ir o f
The to p o f th e abdomen
im m ed iately back o f th e th orax may be o u t a c r o s s w ith s t e r i l e s c is s o r s and
th e r e s u lt i n g f la p formed o f th e e n t ir e d o r s a l w a ll o f th e abdomen may th en
be l i f t e d back w ith a p a ir o f flam ed f o r c e p s , le a v in g th e abdominal v is c e r a
exp osed ."
Glasgow was a b le to g e t good b a c t e r ia l smears from th e caeoae o f
c e r t a in s p e c ie s o f H e te ro p tera , but c o u ld n o t o b ta in c u lt u r e s on ordinary
m ed ia,
ti/ith o th e r s p e c ie s he o b ta in ed lu x u r ia n t growth i n n u t r ie n t b ro th .
A squash v in e in f u s io n b roth composed o f n u t r ie n t b roth and a d eco ctio n
from 160 gra s o f squash v in e per l i t e r was s u c c e s s f u lly u sed i n c u lt i v a t in g
o a eo a l organism s from th e squash bug Anaaa t r i s t i s .
Glasgow found th a t th e
caeoae o f th e Hem iptera o f any one d e f i n i t e s p e c ie s in v a r ia b ly co n ta in ed a
pure c u ltu r e o f b u t one s p e c ie s o f b a cteriu m .
The b a c t e r ia from d if f e r e n t
h o s ts v a r ie d g r e a t l y , from sm all co ccu s l i k e organism s to la r g e s p ir a l
form s.
However, " in w hatever form th e y occu r th ey a re m o rp h o lo g ic a lly
c h a r a c t e r is t ic f o r th e p a r t ic u la r s p e c ie s h arb orin g them."
Glasgow
b e lie v e d th a t th e s e normal b a c te r ia appear n o t o n ly to i n h i b i t th e d evelop ­
ment o f f o r e ig n b a c t e r ia b u t to ex clu d e them a lt o g e t h e r .
This i s a v ery
i n t e r e s t i n g o b s e r v a tio n .
P ie r c e (1 9 2 1 ) g iv e s a summary o f th e reco rd s w hich have been made o f
th e organism s i s o l a t e d from co ck ro a c h e s.
F if t e e n s p e c ie s o f b a c te r ia are
l i s t e d as b ein g e i t h e r i s o l a t e d from th e f e c e s o f t h i s i n s e c t , or th e
a lim en ta ry t r a c t .
P ie r c e a ls o l i s t s 53 s p e c ie s o f b a c t e r ia th a t have been
-8 i s o l a t e d from th e h o u s e - f ly , some from th e s u r fa c e o f th e f l y , and o th e r s
from in t e r n a l o r g a n s.
Much o f the b a c t e r i o lo g ic a l work done on th e house­
f l y has b een w ith abnormal f l o r a .
A ttem pts to e x p e rim en ta lly in tro d u ce
pathogens in t o th e a lim en ta ry t r a c t o f th e f l y and r ec o v er th e v ir u le n t
pathogen from th e f e c e s have been numerous.
S tein h a u s (1941) has made a su rvey o f th e b a c t e r ia l f l o r a o f c e r t a in
i n s e c t s and f in d s " th a t m ost o f th e major ty p es o f b a c t e r ia are r e p r e se n te d .
These in c lu d e g r a m -p o s itiv e and g ra m -n eg a tiv e sh o r t r o d s , g r a m -p o sitiv e
sp ore-form in g b a c i l l i , and g r a m -p o sitiv e c o c c i .
The g ra m -n eg a tiv e sh o r t
rods p redom inate, com p rising s l i g h t l y more than 50 per c e n t o f th e b a c t e r ia l
f l o r a o f th e i n t e s t i n a l t r a c t s o f th e i n s e c t s s tu d ie d ."
He a ls o found "a
number o f b a c t e r ia which w ere e l l i p t i c a l in shape and w hich one i s prompted
to d e s ig n a te as o o o o o b a o ill i.
They v a r ie d in t h e ir p h y s io lo g ic ch a r a c te r ­
i s t i c s as w e ll as in t h e i r r e a c t io n to th e g r a m -sta in .
These form s, however,
may be pleom orphic ty p e s o f th e f a m ilia r sh o r t rods and c o c c i."
A lto g e th e r , S tein h a u s made an in v e s t ig a t io n o f th e b a c t e r ia l f lo r a o f
30 s p e c ie s o f i n s e c t s from 7 ord ers o f th e c l a s s Hexapoda.
He is o l a t e d a
t o t a l o f 83 s t r a in s o f b a c t e r ia , and made s t u d ie s upon them in d e t a il w ith
regard to t h e ir m o rp h o lo g ic, c u lt u r a l, p h y s io lo g ic and p a th o g en ic ch a r a c te r ­
is tic s .
He found th a t o f th e 83 s t r a in s 44 (63 per c e n t) were g ram -n egative
sh o r t r o d s , 17 (2 0 .5 per cen t)w ere g r a m -p o s itiv e spore fo rm ers.
S tein h au s
s t a t e s , how ever, t h a t " th e se p ercen ta g es a re g iv e n m erely f o r th e purpose o f
making approxim ate com parisons and n o t a s th e p ercen ta g es o f th e v a r io u s groups
o f b a c te r ia as they may a c t u a lly be found throughout th e c l a s s Hexapoda."
A lso th a t th e s e f ig u r e s a re s u b j e c t to change depending on th e in t e r p r e t a t io n
-9 made o f th e data p r e se n te d in h is p ap er.
T h irty-tw o o f th e s t r a in s s tu d ie d
did n o t conform to d e s c r ip t io n s g iv e n in B ergey’ s m anual.
were new s t r a in s and 5 were u n id e n t if ie d .
Of t h e s e , 27
From th e 27 new s t r a in s , 21 new
s p e c ie s were e s t a b lis h e d .
F iv e s p e c ie s o f O rthoptera were in c lu d e d in S te in h a u s ' in v e s t ig a t i o n .
B l a t t e l l a g e m a n io a L ., Ueombius f a s o i a t u s , v a r . f a s o ia t u s DeG.,
ConooeophaluB f a s o i a t u s , v a r . f a s o ia t u s DeG., Dlapheromera fem orata Say,
and an u n id e n t if ie d member o f th e T e ttig o n lid a e were th e i n s e c t s which
composed t h i s grou p .
These y ie ld e d 19 s t r a in s o f b a c t e r ia .
F if t e e n o f
th e s e were s t r a in s o f g ram -n egative sh o r t r o d s, 2 were s t r a in s o f gram­
p o s i t i v e c o c c i , and 2 were s t r a in s o f g r a m -p o sitiv e sp o r e -fo r m e r s.
EXPERIMENTAL METHODS
L iv in g specim ens o f G r y llo b la tta oampo d el form! s ca m p o d eifo m is were
c o l l e c t e d in th e F a iry Lake r e g io n o f th e B rldger Range n o rth o f Bozeman,
Montana,
Upon b r in g in g th e specim ens to th e la b o ra to ry th ey were p la ced in
th e r e f r ig e r a t o r so t h a t th ey would s u r v iv e u n t i l needed f o r th e exp erim en t.
TWo a d u lt specim ens were s e le c t e d fo r d is s e c t io n and t r e a t e d as f o llo w s :
The l i v e i n s e c t was p la c e d in a v i a l o f 55 per ce n t a lc o h o l fo r two to th ree
m inutes to remove th e a ir f ilm and a llo w p e n e tr a tio n o f th e d is in f e c t a n t .
S u b seq u en tly i t was p la c e d i n a v i a l o f 1 -1 ,0 0 0 m ercuric c h lo r id e fo r two
to th ree m in u te s, th en in a v i a l o f s t e r i l e w ater and r in s e d w e l l .
Next
the i n s e c t was p la ce d in a s t e r i l e p e t r i d ish w ith a beeswax la y e r in the
bottom .
I t was d is s e c t e d w ith a lc o h o l-fla m e d in str u m e n ts.
A s l i t was made
v e n t r a lly from th e p o s te r io r p a rt o f th e abdomen to th e a n t e r io r part o f th e
-1 0 thorax and th e r e s u lt i n g f la p s pinned to th e s id e s , ex p o sin g th e a lim en tary
tr a c t.
The e n t ir e a lim en ta ry t r a c t was th en removed and d iv id e d in to f o r e ­
g u t, m id -g u t, and h in d -g u t.
Each p o r tio n was p la ced in a s t e r i l e 10 o c .
w ater blank and e m u ls if ie d w ith a s t e r i l e r o d .
The e m u ls if ie d p o r tio n s and
w ater w ere then d is t r ib u t e d among a number o f p e t r i d ish e s and th e d ish e s
poured w ith d e x tr o se a g a r .
Both a e r o b ic and an aerob ic p la t e s o f each
p o r tio n o f th e t r a c t were in cu b a ted a t 4°C, 18°C, and 28°C .
A fte r th ree
days th e p la t e s were examined and i t was found th a t a l l an a ero b ic p la t e s
were d evoid o f grow th.
The a e r o b ic p la t e s in cu b a ted a t 28°C had cop iou s
growth and th o se a t IB0C f a i r grow th .
A fte r a few weeks o f fu r th e r
in c u b a tio n a few c o lo n ie s appeared in th e p la t e s b ein g in cu b a ted a t 4°C.
C o lo n ies were p ick ed from a l l p l a t e s , w ith an attem pt to make a
r e p r e s e n t a t iv e s e l e c t i o n o f th e organism s p r e s e n t, and s tr e a k s were made
on d e x tr o se agar s l a n t s .
A fte r growth was o b ta in ed on th e s la n t s each
c u ltu r e was r e p la te d to f u r t h e r in s u r e pure c u lt u r e s .
The optimum
tem perature f o r a l l b u t one o f th e c u ltu r e s was found to be room tem perature
(2 5 °C ).
One c u lt u r e grew b e s t a t l7°C to 22°C .
From th e m o rp h o lo g ica l s tu d ie s made on t h e s e c u ltu r e s i t became
ap p arent th a t a d iv e r s e group o f organism s was p r e s e n t, and t h a t w ith in
th e li m it a t io n s o f t h i s paper i t would b e w is e to attem p t m erely an
i d e n t i f i c a t i o n o f th e organism s from th e two in d iv id u a ls and to make no
attem p t to determ ine w hether t h is f l o r a i s uniform in a la r g e number o f
sp ecim en s»
M orp h ological s t u d ie s were made in d e t a i l and much tr o u b le was
e x p erien ced due to pleom orphism .
I n o c u la tio n s were made in t o v a rio u s
-1 1 m edia and h ere a g a in s t r a in v a r ia t io n became e v id e n t .
W ith few e x c e p tio n s
a l l in o c u la t io n s were made in d u p lic a te and o f t e n in t r i p l i c a t e or
q u a d r u p lic a te .
In cu b a tio n s were made a t room tem perature e x c e p t w ith g e l a t i n , which
was in cu b a ted i n th e 20°C in c u b a to r .
R egarding in c u b a tio n tim e , sta r c h
h y d r o ly s is t e s t s w ere made a f t e r 5 to 7 d a y s.
6 w eek s.
G e la tin s ta b s w ere in cu b a ted
A ll o th e r media were in cu b a ted f o r a t l e a s t two w eek s.
From tim e to tim e a l l c u lt u r e s were examined fo r p u r ity by r e p la tin g
and making g r a m -s ta in s .
At th e c l o s e o f t h i s in v e s t ig a t i o n a l l c u ltu r e s
in c lu d e d in t h is paper w ere a g a in examined m ic r o s c o p ic a lly from 24 hour
d e x tr o se agar s la n t s and th e morphology checked w ith th e d ata p r e v io u s ly
o b ta in e d f o r each c u lt u r e .
A t o t a l o f 90 s t r a in s o f organism s was o r i g in a lly i s o l a t e d from the
two i n s e c t s .
i n t h i s p ap er.
Of t h e s e , 60 s t r a in s o f E u b a o te r ia le s were f i n a l l y in clu d ed
Most o f th e o th e r s were e lim in a te d as A o tin o ir y o e ta le s,
or o f d o u b tfu l p u r it y ; s e v e r a l d ied a f t e r a few t r a n s f e r s on d ex tro se
aLa r , on w hich th ey grew i n v a ry in g d eg rees a t f i r s t .
f i r s t i n s e c t examined a re la b e l le d ' A '•
UbeUsi 1B1
C u ltu res from the
Those from th e secon d in s e c t are
-12
RESULTS
The f i f t y organism s is o la t e d from th e a lim en ta ry t r a c t o f
C r y llo b la tta cam ^odeiform is a re l i s t e d below w ith th e m o rp h o lo g ica l and
p h y s io lo g ic a l c a r a c te r s found by th e a u th o r . The r e s u l t s recorded wore
c o n f i n e d by rep ea ted t e s t s , and in th o s e c a s e s where th e y d id n o t check
th e t e r i v a r ia b le was u s e d .
Al
Iicro co o cus s p .
I o c c ii 0 .6 t o 1 .0 m icron in d ia m eter , o ccu rrin g s i n g l y , in p a ir s ,
and in m a sse s. E o n -m o tile . G r a m -p o sitiv e.
C e la ti s t a b i S tr a tifo r m li q u e f a c t io n .
' o x tr o s e agar s l a n t i Abundant, s p r e a d in g , r a is e d , g l i s t e n i n g , sm ooth,
p in k is h .
Litmus n i l ; S lo w ly becoming a l k a l i n e .
Iydrogen s u l f i d e p ro d u ctio n s l i g h t .
K oser’ s c i t r a t e p o s i t i v e .
S tarch h y d r o ly s is p o s i t i v e .
I i t r i t e s produced from n i t r a t e s .
In dol n o t produced.
ogoe-P r o skauor n e g a t iv e .
Acid from d e x t r o s e . A cid may or ay n o t be produced from m a lto se .
o
a o i from l a c t o s e , su c r o se or m a n n ito l.
P otato t S ca n ty , e o h in u la t e , r a is e d , g l i s t e n i n g , sm ooth, p in k ,
medium n o t darkened.
A ero b ic.
a b l t a t i P o r e -g u t.
A2
Iicroooccu s s p .
C oooii 0 . 8 to 1 .4 m icrons in d ia m e te r , o ccu rr in g s i n g l y , in .a ir s and
in m a sse s, C rai- p o s i t i v e . M on-m otile.
G e la tin s t a b i ' ap iform li q u e f a c t io n .
D extrose a, ar s l a n t i Abundant, f i l i f o r m , co n v ex , g l i s t e n i n g , sm ooth,
oreamy y e llo w .
Litmus m ilk i S lo w ly becoming w eak ly a l k a l i n e .
hydrogen s u l f i d e v a r ia b le .
K oser’s c i t r a t e p o s i t i v e .
S tarch n o t h y d ro ly ze d .
N i t r i t e s produced from n i t r a t e s .
In dol n o t produced.
V oges-Proekauer n e g a t iv e .
Acid ir d e x tr o s e and a l t o s e , Po a c id from s u c r o s e , la c t o s e or
m a n n ito l.
P o ta to : S ca n ty , e o h in u la t e , f l a t , g l i s t e n i n g , sm ooth, y e llo w , medium
unchan e d .
A erobic *
KabI t a t ; Por e - g u t .
B
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-1 5
AS
Iic ro c o c c u s sp»
C o co ii 1 .1 t o 1*4 TTiierons in d ia n e t e r , o ccu rr in g s i n g l y , in p a ir s ,
and in m s s s s . ] o n - m o t ile . G r a m -p o sitiv e.
G e la tin s t a b ; S tr a tifo r m , n ap iform or ln fu n d ib u lifo r m li q u e f a c t io n ,
e x Urose a^.ar s la n t : Abundant, f i l i f o r i, r a is e d , g l i s t e n i n g , sm ooth,
y e llo w .
Litmus iiilk : S lo w ly becoming a llc a lin e , reduced in bottom .
ydrogen s u l f i d e n e g a t iv e .
K oser's c i t r a t e p o s i t i v e .
S tarch h y d ro ly ze d •
i t r i t e s produced from n i t r a t e s .
In d ol n o t produced.
Voj e s -P ro s’ auer n e g a t iv e .
Vo a c id fro : ca rb o h y d ra tes.
Potato I
© d erate, sp r e a d in g , r a i s e d , d u l l , s l i g h t l y v e r r u c o s e , d u ll
y e llo w , medium d arkened.
A ero b ic.
a b i t a t j F o r e -g u t.
A4
ic r o c o c c u s s u b fla v u s
C o cci: 0 .8 to 1 .6 m icrons in d ia m e te r , o ccu rrin g s i n g l y , in p a ir s , and
in m a sses. Ko n - m o t ile . G r a m -p o sitiv e.
G e la tin sta b * L iq u e fa c tio n n a p ifo r i o r s t r a t if o r m .
D extrose agar s l a n t : Abundant, f i l i f o r m , co n v ex , g l i s t e n i n g , sm ooth,
p ale y e llo w .
Litmus i l l ': S lo w ly becoming a l k a l i n e ,
hydrogen s u l f i d e n o t form ed.
Kosar's c i t r a t e p o s i t i v e .
S tarch h y d r o ly s is v a r i a b l e .
i t r i t e s n o t produced from n i t r a t e s .
In dol n o t produced.
V oges-P roskauer n e g a t iv e .
Fo a c id from d e x t r o s e , m a lto se , l a c t o s e , s u c r o s e , or aim I t o l .
P o ta to : S ca n ty , f i l i f o r m , co n v ex , d u l l , sm ooth, y e llo w , nedium
unchanged.
A ero b ic.
H a b ita t * F o r e -g u t.
-14'
AS
lc ro c o c c u s e p id e r iild ls
C o o cit 0» t o 1#4 m iorons in d ia m e te r , occu rrin g s i n g l y , in p a ir s
and in M asses* G ra m -p o sitiv e * N on-m otlie*
G e la tin sta b t Ne liq u e f a c t io n *
D ex tro se agar s l a n t t M oderate, e o h in u la t e , r a is e d , g l i s t e n i n g ,
sm ooth, w h ite*
Litmus m ilk : S lo w ly becoming a cid * No curd*
ydrogen s u l f i d e n o t formed*
K oser*6 c i t r a t e v a r ia b le *
S tarch n o t h ydrolyzed*
i t r a t e s reduced t o n i t r i t e s •
Indol n o t produced*
V oges-P roskauer n e g a t iv e .
Acid in l a c t o s e , s u c r o s e , d e x t r o s e , and m a lto s e , no a cid in m annitol*
P o ta to : S ca n ty , e o h in u la t e , r a i s e d , d u l l , s l i g h t l y r u g o s e , g ra y ,
medium n o t darkened*
A erobic »
H a b ita t i F ore-gu t*
AS
M icrococcus ep id erm ld is
C o cci; 0*4 t o 0 .8 m icron in d ia m eter , o ccu rrin g s i n g l y , in p a ir s , and
in m asses* G ram -p ositive* lo n - m o t ile .
G e la tin sta b s No liq u e f a c t io n *
D extrose a, ar s l a n t : M oderate, e o h in u la t e , r a is e d , g l i s t e n i n g , sm ooth,
w h it e •
Litmus m ilk : S lo w ly becoming a c id * Ho c o a g u la tio n *
M yrogen s u l f i d e n o t produced*
K oser' s c i t r a t e n e g a tiv e *
S tarch n o t hydrolyzed*
M t r i t e s produced from n it r a t e s *
In d ol n o t produced*
V oges-Proekauer n e g a tiv e *
Acid from d e x t r o s e , m a lto s e , s u c r o se and la c to s e * No a c id from
m an n itol*
P o ta to : S ca n ty , e o h in u la t e , r a i s e d , g l i s t e n i n g , sm ooth, g r a y is h -w h ite ,
medium n o t d is c o lo r e d *
A e r o b ic , f a c u lt a t iv e *
h a b ita t: F ore-gu t*
-1 5
A7
^ icro o o co u s ep ld er m id is
C o c e ii 0*2 to 0*7 m icrons in d ia m eter , o ccu rr in g s i n g l y and in p a irs*
Gra - j o s it iv o * Hon-cmatile*
G e la tin s t a b : Ko liq u e f a c t io n *
D extrose a;;ar s la n t * S ca n ty , b ead ed , co n v ex , g l i s t e n i n g , sm ooth,
w h it e .
Litmus m ilk s S lo w ly becoming w eak ly a c i d .
Hydrogen s u l f i d e n o t produced.
K ocer’s c i t r a t e p o s i t i v e .
S tarch n o t h yd rolyzed*
K itr a te s reduced t o n i t r i t e s .
In d o l n o t produced*
V oges-P roskauer n e g a t iv e .
Acid from d e x t r o s e , s u c r o s e , l a c t o s e , and m a lto s e . To a c id from
m a n n ito l.
P o t a t o t S ca n ty , e c h in u la t e , r a i s e d , s l i g h t l y d u l l , s l i g h t l y v e r r u c o sa ,
l i g h t g r a y , p o ta to unchanged.
A erobic *
H ab itat* F o r e -g u t.
AS
ic r o c o c c u s s p .
C occi* 0 .5 to 0*4 m icron i n d ia m e te r , o ccu rrin g s in g ly and in p a ir s .
G ram -p ositive* I o n -m o tile*
G e la tin sta b * S tr a tifo r m li q u e f a c t io n •
D extrose a ar s la n t* M oderate, e c h in u la t e , f l a t , g l i s t e n i n g , sm ooth,
grayish -b row n •
Litmus m ilk *
ed u ctio n and curd in low er part* P ep ton ized in upper
part*
Hydrogen s u l f i d e n o t produced*
K oser's c i t r a t e n e g a tiv e *
S tarch n o t h y d r o ly z e d .
T i t r i t e s produced from n i t r a t e s .
In d o l n o t produced.
Vogas-ProsH auer n e g a t iv e •
Acid from l a c t o s e , s u c r o s e , d ex tro se* Io a c id from m an n itol*
P otato*
b d e r a te , sp r e a d in g , r a i s e d , g l i s t e n i n g , sm ooth, g ra y .
A erobic *
H a b ita t * F o re-g u t*
16'
B9
lc r o c o c c u e sp*
CoeclT 0*8 t o 1*4 m icrons in d ia m e te r , o ccu rrin g s i n g l y
Ir
p a ir s and snassas* G ram -p ositive* io n -m o tile *
G e la tin s t a b t S tr a tifo r m liq u e f a c t io n *
D extrose agar s l a n t t M oderate, e c h in u la t e , r a is e d , g l i s t e n i n g ,
sm ooth, cream co lo red *
Litmus m ilk j S lo w ly becoming a lk a lin e w ith r e d u c tio n in bottom .
Hydrogen s u l f i d e n o t produced.
K oser*s c i t r a t e p o s it iv e *
S tarch n o t hyd rolyzed*
T it r a te s reduced t o n i t r i t e s *
In d ol n o t produced*
Voges -Proslcauer n e g a tiv e »
A ero b ic, f a c u lt a t iv e *
H a b ita ti F o r e -g u t.
A cid in d e x t r o s e , s u c r o s e , l a c t o s e , m altose. Ko a c id in m annitol*
AlO
M icrococcus sp*
C o c c ii 1 .1 t o 1*6 m icrons in d ia m e te r , o ccu rrin g s i n g l y , in p a ir s
and in m asses* lo n - m o t il e . G r a m -p o sitiv e.
G e la tin sta b * Ho li q u e f a c t io n .
D extrose a ar s la n t * M oderate, e c h in u la t e , r a i s e d , g l i s t e n i n g ,
sm ooth, l i g h t o ra n g e.
Litmus m ilk i S lo w ly becoming a lk a lin e *
Hydrogen s u l f i d e n o t form ed.
K oser*s c i t r a t e v a r ia b le .
S tarch h y d r o ly s is v a r ia b le .
i t r a t e s reduced t o n i t r i t e s *
In d ol n o t p roduced.
V ogus-Proskauer n e g a t iv e .
P o ta to : Ho grow th .
A ero b ic.
■oak a c id produced from d e x t r o s e , l a c t o s e , s u c r o s e , m a lt o s e . Ko
a c id from m a n n ito l.
-17.
A ll
M iorooooous s p .
C oooii 0 .6 to 1 .0 m ioron in diam eter o ccu rr in g s i n g l y , i n p a ir s
and in m a sse s. N o n -m o tile . G r a m -p o sitiv e.
G e la tin s ta b : S tr a tifo r m li q u e f a c t io n .
D extrose a^ar s l a n t i M oderate, b ead ed , r a is e d , g l i s t e n i n g , smooth,
g r a y is h -w h ite .
Litmus m ilk i S low ly becom ing a lk a li n e , reduced in bottom .
hydrogen s u l f i d e n o t produced,
K oser1K c i t r a t e p o s i t i v e .
S ta rch h y d r o ly s is v a r ia b le .
N itr a te s reduced to n i t r i t e s .
In d o l n o t produced.
V oges-Proskauer n e g a t iv e .
A cid from d e x tr o s e , s u c r o s e , m a lto se and l a c t o s e . No a c id from
m a n n ito l.
P o t a t o i S ca n ty , e o h in u la t e , f l a t , g l i s t e n i n g , sm ooth, oran ge,
medium unchanged.
A ero b ic.
H a b ita t: P o r e -g u t.
B12
M icrococcus s p .
C occi i 0 .9 to 1 .4 m icrons in d ia m eter, occu rrin g s i n g l y , in p a ir s and
in m a sse s. G r a m -p o sitiv e. N o n -m o tile.
G e la tin a t a b i Napiform l i q u e f a c t io n .
D extrose agar s l a n t t M oderate, e o h in u la t e , r a is e d , d u l l , smooth,
crea m -co lo red .
Litmus m ilk i S lo w ly becoming w eakly a lk a li n e .
Ifydrogen s u l f i d e n o t produced.
K oser1s c i t r a t e p o s i t i v e .
S ta rch n o t h y d r o ly z e d .
N i t r i t e s produced from n i t r a t e s .
In d ol n o t produced.
V oges-P roskauer n e g a t iv e .
A cid from d e x tr o s e , s u c r o s e , m a lto s e , and l a c t o s e . No a c id from
m a n n ito l.
A ero b ic.
H a b ita t: H ind-gut
16AlS
M icrooooous sp«
C oooit 0 ,2 to 0 ,7 m icron in d ia m eter, o ccu rr in g s i n g l y , i n p a ir s , and
in m a sse s. Gra m - p o s it iv e . N o n -m o tile.
G e la tin s t a b : S tr a tifo r m li q u e f a c t io n .
D ex tro se agar s la n t : Abundant, sp r e a d in g , r a is e d , g l i s t e n i n g , sm ooth,
tr a n sp a r e n t w h it e .
Litmus m ilk I S low ly becoming p e p to n iz e d .
Hydrogen s u l f i d e n o t produced.
K osert S c i t r a t e p o s i t i v e .
S ta rch h y d r o ly s is s l i g h t .
N it r a t e s n o t reduced to n i t r i t e s .
In d o l n o t produced.
V oges-Iro sk a u er n e g a t iv e .
Weak a c id p ro d u ctio n from d e x tr o s e . No a c id from m a lto s e , la c t o s e ,
s u c r o s e , or m a n n ito l.
P o t a t o i Abundant, s p r e a d in g , r a is e d , g l i s t e n i n g , sm ooth, d ir ty g ra y ,
medium darkened.
A ero b ic.
Habi t a t i F o r e -g u t.
A l4
M icrococcus s p .
C occi i 0 .7 to 1 .6 m icrons in d ia m eter, o ccu rrin g in p a ir s . Nonm o t il e . G r a m -p o sitiv e.
G e la tin s t a b i S tr a tifo r m li q u e f a c t io n .
D extrose agar s l a n t i Abundant, e o h in u la t e , co n v ex , g l i s t e n i n g ,
sm ooth, g r a y is h -w h ite .
Litmus m ilk i S low ly becoming a l k a l i n e , reduced in bottom .
Hydrogen s u l f i d e v a r ia b le .
K oser’ s c i t r a t e p o s i t i v e .
S tarch n o t h y d ro ly ze d .
N i t r i t e s n o t produced from n i t r a t e s .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
No a c id i n c a rb o h y d ra tes.
P o t a t o : No grow th.
A ero b ic.
H abit a t t F o r e -g u t.
19A16
Mlorooooous s p .
C oooii 0 .3 to 1 .2 m icrons In d ia m eter , o ccu rr in g s in g ly and in m a sse s.
O r a m -p o sitiv e. M on -m otile.
G e la tin sta b i S tr a tifo r m l i q u e f a c t io n .
D extrose agar s la n t : Abundant, e o h in u la t e , r a is e d , g l i s t e n i n g ,
sm ooth, w h ite .
Litmus m ilk : S low ly becoming a o id . Reduced in low er tw o -th ir d s .
S o f t curd on bottom .
Hydrogen s u l f i d e n o t form ed.
K oser’ s c i t r a t e p o s i t i v e .
S taroh h y d ro ly zed s l i g h t l y .
N itr a te s n o t red u ced .
In d o l n o t produced.
V oges-Proakauer n e g a t iv e .
Aoid in d e x tr o s e , l a c t o s e , s u c r o s e , m a lto s e , and m a n n ito l.
P o ta to : Abundant, s p r e a d in g , r a i s e d , g l i s t e n i n g , sm ooth, d ir t y g ra y ,
medium darkened s l i g h t l y .
A ero b ic.
H a b ita t: F o r e -g u t.
AlS M iorocooous fr e u d e n r e io h ii
C o o o i: 0 .6 i n d ia m eter , o ccu rr in g s i n g l y , in p a ir s , and in m a sses.
N o n -m o tile. G r a m -p o sitiv e.
G e la tin s ta b : I n fu n d ib u lifo r a li q u e f a c t io n .
D extrose agar s la n t : S ca n ty , f i l i f o r m , sm ooth, r a i s e d , g l i s t e n i n g ,
w h ite .
Litmus m ilk : S lo w ly becoming a o id .
Hydrogen s u l f i d e n e g a t iv e .
S tarch h y d r o ly s is n e g a t iv e .
K oserf S c i t r a t e p o s i t i v e .
N itr a te s n o t red u ced .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
Aoid in d e x tr o s e , l a c t o s e , s u c r o s e , m a lto s e , and m a n n ito l.
P o ta to : Abundant, sp rea d in g , r a i s e d , sm ooth, d u l l , g r a y is h c e n te r ,
l i g h t orange e d g e s , p o ta to darkened.
A erobic f a c u l t a t i v e .
H a b ita t: F o r e -g u t.
20A l7
Haoterium s p .
S h ort r o d s i Very p leo m o rp h ic. Some la r g e and p ea r-sh a p e d . 0 .6 by
1 .1 m icr o n s, o ccu rr in g s in g ly and in p a ir s . A sporogenous. G ra m -p o sitiv e.
N o n -m o tile.
G e la tin sta b s No li q u e f a c t io n .
D extrose agar s la n t s M oderate, e c h in u la t e , r a is e d , g l i s t e n i n g , sm ooth,
g r a y is h -w h ite .
Litmus m ilk s S lo w ly becoming a l k a l i n e ,
ffydrogen s u l f i d e n o t produced.
K oser’ s c i t r a t e v a r ia b le .
S ta rch n o t h y d r o ly se d .
N it r a t e r e d u c tio n v a r ia b le .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
Acid from d e x tr o s e , l a c t o s e , m a lto s e , s u c r o s e , or m a n n ito l.
P o ta to I S ca n ty , e c h in u la t e , r a is e d , g l i s t e n i n g , sm ooth, g ra y , medium
unchanged.
A ero b ic.
H ab itats P o r e -g u t.
AlS Sac herium s p .
R odss Very p leom orp h ic. 0 .8 to 1 .6 m icro n s, o c c u r r in g s in g ly , and
i n p a ir s . G r a m -p o sitiv e . N o n -m o tile. A sporogenous.
G e la tin sta b s No li q u e f a c t io n .
D extrose agar s la n t s Abundant, b ead ed , con vex, g l i s t e n i n g , smooth,
cre a m -c o lo red .
Litmus m ilk s S lo w ly becoming a lk a li n e in upper p o r t io n , reduced
b elo w .
Hydrogen s u l f i d e n e g a t iv e .
K oser’ s c i t r a t e v a r i a b l e .
S ta rch n o t h y d ro ly ze d .
N itr a te s reduced to n i t r i t e s .
In d o l n o t produced.
V oges-Proskauer n e g a t iv e .
Aoid from d e x tr o se and s u c r o s e . No a c id in m a lto s e , l a c t o s e , or
m a n n ito l.
F o ta to s S ca n ty , e c h in u la t e , r a i s e d , g l i s t e n i n g , sm ooth, g ra y , medium
n o t changed.
A ero b ic.
H a b ita ts F o r e -g u t.
A l9
up.
plwom orphio* O.S m ioron w ide by 1 .1 to 1 .7 m iorons lon«;,
o o o u rrin r o^o^ly nnd in om lr*. fJrwm -poaitiTe. Non-mo t i l e . Aaporogenous.
G elfttln atftbt S tr a tifo r m li q u e f a c t io n .
D ex tro se agar s l a n t « Abundant, f i l i f o r m , r a is e d , g l i s t e n i n g , sm ooth,
Crear^r-W hite c o l o r .
Litmus m ilk j S low ly becom ing p e n to n ie e d .
rt’-drogen a u lf id e n e g a t iv e .
R oger's c i t r a t e p o s i t i v e .
S tarch h y d r o ly se d .
N itr a te s reduced t o n i t r i t e s .
In d o l n o t p rod u ced .
V oges-Proekauer n e g a t iv e .
A cid in d e x tr o se and s u c r o s e . No a o id in l a c t o s e , m a lto s e , or m an n itol
P o t a t o i Abundant, sp r e a d in g , r a i s e d , g l i s t e n i n g , sm ooth, g r a y is h brown c o l o r , medium darkened.
A ero b ic.
H a b ita t; F o r e -g u t.
B20
Baoterium s p .
Sm all rods* 0 .3 to 0 .5 m icron w ide by 0 .8 to 1 .6 m icrons lo n g ,
o ccu rr in g s i n g l y , in p a ir s and sh o r t c h a in s . G r a m -p o sitiv e. N o n -m o tile.
A eporogenous.
G e la tin stab* S tr a tifo r m li q u e f a c t io n .
D extrose agar s la n t* Abundant, sp r e a d in g , r a is e d , g l i s t e n i n g , sm ooth,
w h ite .
Litmus m ilk* S lo w ly becoming p e p to n iz e d .
'tydrogen s u l f i d e n e g a t iv e .
K oser'e c i t r a t e p o s i t i v e .
S tarch n o t h y d r o ly z e d .
N itr a te r e d u c tio n v a r ia b le .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
A cid i n d e x tr o se and s u o r o se . No a c id in la c t o s e , m a lto s e , or m an n itol
P o ta to * Abundant, sp r e a d in g , r a i s e d , g l i s t e n i n g , sm ooth, d ir ty g ra y ,
medium darkened.
A ero b ic.
H ab itat* H in d -g u t.
22B21
Baoterimn ap .
S h ort rod s i P leom orp h lo. 0 .7 to 0 .8 miorOn w ide by 0 .8 to 1 .7 m icrons
lo n g , ooou rrin g s in g ly and i n p a ir s . G r a m .p o sitiv e . N o n -m o tile.
A eporogenous.
G e la tin sta b s S tr a tifo r m li q u e f a c t io n .
D extrose agar s la n t s Abundant, sp rea d in g , con vex, g l i s t e n i n g , sm ooth,
cream y-w h ite.
Litamis m ilk s S lo w ly becoming a lk a li n e w ith r e d u c tio n in bottom .
hydrogen s u l f i d e p ro d u ctio n v a r ia b le .
Kosort s c i t r a t e p o s i t i v e .
S tarch h y d ro ly ze d .
N i t r i t e s produced from n i t r a t e s .
In d o l n o t produced.
V oges-Proskauer n e g a t iv e .
A cid from d e x tr o se and s u c r o s e . Weak a c id from m a n n ito l. No a o id in
m a lto s e , or l a o t o s e .
P o ta to s Abundant, sp rea d in g , r a i s e d , d u ll, sm ooth, d ir t y g ra y , medium
darkened.
A ero b ic.
H ab itats P o r e -g u t.
A22
Bao terium o p .
S h ort rod s i 0 .3 to 0 .6 m icron w ide by 0 .6 to 1 .4 3 miorono lo n g ,
o ccu rr in g s in g ly and in p a ir s . G r a m -p o sitiv e . N o n -m o tile. Asporogenous.
G e la tin sta b s S tr a tifo r m li q u e f a c t io n .
D ex tro se agar s la n t s Abundant, e o h in u la t e , r a i s e d , g l i s t e n i n g , sm ooth,
g r a y is h -w h ite .
Litmus m ilk s S lo w ly becoming p e p to n iz e d in upper h a l f o f tu b e,
reduced in low er p o r tio n w ith r o f t cu rd .
Hydrogen s u l f i d e p o s i t i v e .
K osert B o i t r a t e p o s i t i v e .
S tarch s tr o n g ly h y d r o ly z e d .
N i t r i t e s produced from n i t r a t e s .
In d ol n o t produced.
V oges-Proskauer n e g a t iv e .
Aoid in d e x tr o s e , s u c r o s e , l a c t o s e . No a o id i n m a lto se or m a n n ito l.
P o ta to s Abundant0 sp r e a d in g , r a i s e d , d u l l , smooth, g r a y ! s h -brown,
medium darkened.
A ero b ic.
H a b ita ts F o r e -g u t.
-2 3 B23
Bacterium sp«
Sm all r o d s i 0 .3 to 0 .5 mioron w ide by 0 .8 to 1 .4 m icrons lo n g ,
o ccu rr in g s in g ly and i n p a ir s . G r a m -p o sitiv e. N o n -m o tile. A sporogenous.
G e la tin s ta b : S tr a tifo r m li q u e f a c t io n .
d ex tro se agar s l a n t : Abundant, sp r e a d in g , r a is e d , g l i s t e n i n g ,
sm ooth, creamy w h it e .
Litmus m ilk : S lo w ly becoming p ep to n ized i n upper p o r t io n . S o ft curd
and reduced b elo w .
Hydrogen s u l f i d e p ro d u ctio n v a r ia b le .
K oser1S c i t r a t e p o s i t i v e .
S ta rch h y d r o ly z e d .
N itr a te s reduced to n i t r i t e s .
In d o l n o t produced.
Voge s - Proskauer n e g a t iv e .
A cid from d e x tr o s e . A cid p ro d u ctio n v a r ia b le from s u c r o s e . No a c id
produced from l a c t o s e , m a lto s e , or m a n n ito l.
P o ta to : Abundant, s p r e a d in g , co n v ex , sm ooth, g l i s t e n i n g , d ir t y
g r a j , medium unchanged.
A ero b ic.
H a b ita t: H in d -g u t.
A24
Bacterium s p .
S h ort r o d s : 0 .8 4 to 1 .1 2 m icrons w ide by 1 .4 to 1 .6 m icrons lo n g ,
o c c u r r in g s in g ly and in p a i r s . Gra m - p o s it iv e . N o n -m o tile. A sporogenous.
G e la tin s ta b : S tr a tifo r m li q u e f a c t io n .
D extrose agar s l a n t : Abundant, s p r e a d in g , r a is e d , g l i s t e n i n g , sm ooth,
w h it e .
Litmus m ilk : S lo w ly becom ing p ep to n ized w ith r e d u c tio n i n bottom .
Hydrogen s u l f i d e v a r ia b le .
K oser1s c i t r a t e p o s i t i v e .
S tarch h y d r o ly z e d .
N itr a te s n o t red u ced .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
No a o id in m a n n ito l, l a c t o s e , d e x tr o s e , m a lto se , or s u c r o s e .
P o ta to : Abundant, sp r e a d in g , r a i s e d , g l i s t e n i n g , sm ooth, d ir ty
gray medium darkened.
A ero b ic.
H a b ita t: F o r e -g u t.
24A26
Daoterium s p .
S h ort r o d st 0 .5 to 0 ,6 m icron wide by 0 ,5 to 1 ,0 m ioron lo n g ,
ooou rrin g s in g ly and in p a i r s , N o n -m o tile . G ra m -p o sitiv e. A sporogenous.
G e la tin sta b t S tr a tifo r m l i q u e f a c t io n .
D ex tro se agar s l a n t : Abundant, sp r e a d in g , r a is e d , g l i s t e n i n g , sm ooth,
cream c o lo r e d .
Litmus m ilk : S lo w ly becoming p ep to n ized i n upper p o r t io n , lower
p a r t red u ced .
Hydrogen s u l f i d e n e g a t iv e ,
Koaer1B c i t r a t e p o s i t i v e .
S tarch h y d r o ly s is v a r i a b l e .
N i t r i t e s n o t produced from n i t r a t e s ,
In d o l n o t produced,
V oges-P roskauer n e g a t iv e ,
Aoid from d e x tr o s e and s u c r o s e . No a c id from m a lto s e , la c t o s e , or
m a n n ito l.
P o ta to : Abundant, s p r e a d in g , r a i s e d , g l i s t e n i n g , sm ooth, d ir t y g ra y ,
medium darkened.
A ero b ic.
H a b ita t: F o r e -g u t,
B26
Bacterium s p .
Sm all r o d s : 0 ,3 to 0 .7 m icron w ide by 1 ,1 to 1 .5 m icrons lo n g ,
ooou rrin g s in g ly and i n p a i r s . G r a m -p o sitiv e . N o n -m o tile . A sporogenous.
G e la tin s ta b : S tr a tifo r m li q u e f a c t io n .
D extrose agar s l a n t : Abundant, sp r e a d in g , con vex, g l i s t e n i n g ,
sm ooth, creaky w h it e .
Litmus m ilk : S lo w ly p ep to n ized i n upper p o r tio n w ith r e d u c tio n
b elow .
Hydrogen s u l f i d e p o s i t i v e .
K o ser's c i t r a t e p o s i t i v e .
S tarch s tr o n g ly h y d r o ly z e d .
N itr a te s n o t red u ced .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
Aojd from d e x tr o s e , l a c t o s e , s u c r o s e , and m a n n ito l. No a c id from
m a lto s e .
P o ta to : Abundant, s p r e a d in g , r a i s e d , g l i s t e n i n g , sm ooth, d ir ty
gray medium darkened.
A ero b ic.
H a b ita t: F o r e -g u t.
25
A27
Bao terium s p .
R odsi P leom orp h ic. 1 .0 to 1 .3 m iorons w ide by 2 .0 m icrons lo n g ,
o c c u r r in g s i n g l y , and i n s h o r t c h a in s . G r a m -p o sitiv e. M o tile .
A sporogenous.
G e la tin s t a b i C rateriform li q u e f a c t io n .
D extrose agar s la n t : M oderate, e o h in u la t e , r a is e d , g l i s t e n i n g ,
sm ooth, g r a y is h -w h ite .
Litmus m ilk i S lo w ly becoming a l k a l i n e .
Rydrogen s u l f i d e n o t produced.
K o ser's c i t r a t e p o s i t i v e .
S ta rch n o t h y d r o ly se d .
N it r a t e s n o t red u ced .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
Weak a c id from d e x tr o s e , s u c r o s e , and m a lto s e . No a c id from la c t o s e
or m a n n ito l.
P o t a t o i M oderate, s p r e a d in g , r a i s e d , g l i s t e n i n g , sm ooth, g ra y ,
medium darkened.
A ero b ic.
H a b ita ti P o r e -g u t.
A28
Bacterium s p .
R odsi 0 .3 t o 0 .6 m icron wide by 1 .0 to 1 .4 m icrons lo n g , o ccu rrin g
s in g ly and i n p a ir s . M o tile . G r a m -p o sitiv e. Asporo g en o u s.
G e la tin s t a b i S tr a tifo r m li q u e f a c t io n .
D extrose agar s l a n t i M oderate, e o h in u la t e , r a is e d , g l i s t e n i n g ,
smooth, g r a y is h - w h it e .
Litmus m ilk : S low ly p e p to n ise d in upper th r e e -q u a r te r s o f tu b e,
s o f t curd and reduced b elo w . D u p lic a te a lk a li n e in upper h a l f , reduced
i n th e low er h a l f .
Rydrogen s u l f i d e n o t form ed.
K osert S c i t r a t e p o s i t i v e .
S tarch h y d ro ly ze d .
N i t r i t e s n o t produced from n i t r a t e s .
In d ol n o t produced.
V oges-P roskauer n e g a t iv e .
A cid produced i n d e x tr o s e , s u c r o s e ,m a n n ito l, and m a lt o s e . No a c id
in l a c t o s e .
P o ta to : M o d era te ,•sp rea d in g , r a i s e d , d u ll, v e r r u o o s e , y e llo w , medium
darkened.
A ero b ic.
H a b ita t: P o r e -g u t.
-2 6 A29
Baoterium sp#
R odsi 0 .2 to 0 ,6 m ioron w ide by 1 .4 to 2 .2 m icrons lo n g , occu rrin g
s in g ly and i n p a ir s . M o t ile . G r a m -p o sitiv e. A eporogenous.
G e la tin s ta b : S tr a tifo r m li q u e f a c t io n .
D extrose agar s l a n t : Abundant, sp r e a d in g , r a is e d , g l i s t e n i n g ,
smooth, d u ll y e llo w .
Litmus m ilk : P e p to n iz a tio n i n upper t w o -th ir d s , low er p a r t reduced
and s o f t curd form ed.
% dro gen s u l f i d e p o s i t i v e .
K oser1S c i t r a t e p o s i t i v e .
S ta rch n o t h y d r o ly se d .
N i t r i t e s produced from n i t r a t e s .
In d o l n o t produced.
V oges- Proskauer n e g a t iv e .
No a c id from ca rb o h y d ra tes.
P o ta to : Abundant, sp rea d in g , r a is e d , g l i s t e n i n g , sm ooth, d ir ty
oran ge, medium darkened.
A e r o b ic ,f a c u lt a t iv e .
Habit a t : F o r e -g u t.
A30
B a cter iu m s p .
Rods: 0 .2 to 0 .3 m ioron wide by 0 .9 to 1 .6 m icrons lo n g , o ccu rrin g
m o stly s i n g l y , some i n p a i r s . G r a m -p o sitiv e. M o t ile . A sporogenous.
G e la tin s ta b : S tr a tifo r m li q u e f a c t io n .
D ex tro se agar s la n t : Abundant, sp r e a d in g , r a is e d , g l i s t e n i n g ,
sm ooth, d u ll y e llo w .
Litmus m ilk : Rapid p e p to n iz a tio n f i n a l l y throughout e n t ir e tu b e .
Hydrogen s u lf id e produced.
K oser’ s c i t r a t e p o s i t i v e .
S ta rch n o t h y d ro ly ze d .
N itr a te s reduced to n i t r i t e s .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
No a c id from ca rb o h y d ra tes.
P o ta to : Abundant, sp rea d in g , r a is e d , g l i s t e n i n g , sm ooth, d ir ty
o ra n g e, darkened.
A ero b ic, f a c u l t a t i v e .
H a b ita t: F o r e -g u t.
-2 7 A31
Baoterium s p .
Sm all rods i 0 .2 m icrons w ide toy 0 .8 to 1 .4 m icrons lo n g , o ccu rrin g
s i n g l y . G r a m -p o sitiv e. M o t ile . A sporogenous.
G e la tin s t a b i S tr a tifo r m li q u e f a c t io n .
D extrose agar s la n t ! M oderate, e o h in u la t e , r a is e d , g l i s t e n i n g ,
sm ooth, g r a y is h -w h ite .
Litmus m ilk i S lo w ly becoming a l k a l i n e , reduced i n bottom .
Hydrogen s u l f i d e n e g a t iv e .
K oser1s c i t r a t e v a r ia b le .
S ta rch n o t h y d ro ly ze d .
N it r a t e s n o t red u ced .
In d o l n o t produced.
V oges-Proskauer n e g a t iv e .
No a c id from ca rb o h y d ra tes.
P o t a t o i Abundant, sp r e a d in g , r a i s e d , g l i s t e n i n g , sm ooth, g r a y ish brown, medium darkened.
A ero b ic.
H a b ita ts F o r e -g u t.
AS2
Bacterium s p .
Hods s 0 .2 to 0 .7 m icron w id e by 0 .8 to 1 .4 3 m icrons lo n g , o ccu rrin g
s in g ly and in p a ir s . A sporogenous. G r a m -p o sitiv e. N o n -m o tile.
G e la tin s ta b : No li q u e f a c t io n .
D extrose agar s la n t s Abundant, s p r e a d in g , r a is e d , g l i s t e n i n g ,
sm ooth, p in k is h .
Litmus m ilk s I n e r t .
Ttydrogen s u l f i d e n o t produced.
K oser's c i t r a t e n e g a t iv e .
S taroh h y d ro ly ze d .
N i t r i t e s n o t produced from n i t r a t e s .
In d o l n o t produced.
V ogee-Proskauer n e g a t iv e .
Aoid i n d e x tr o s e , m a lto s e , and l a c t o s e . No a c id in su c r o se or
m a n n ito l. No a c id i n r a f f i n o s e .
.
P o ta to s S l i g h t , beaded to sca n ty f i l i f o r m , r a is e d , g l i s t e n i n g ,
smooth, y e llo w , medium n o t d is c o lo r e d .
Twaperature r e l a t io n s s Grows a t room tem p eratu re. Grows b e s t a t
17°C to 22°C .
A ero b ic.
H a b ita ts F o r e -g u t.
-2 8 A33
A ero b a o illu s Group
Large r o d s i 0 .6 to 1 .0 m icron w id e by 2 .1 to 3 .4 m icrons lo n g ,
o c c u r r in g s i n g l y , in p a ir s , and in sh o r t c h a in s . Spores c e n t r a l , e lo n g a t e .
Sporan gia c y l in d r ic a l to s l i g h t l y b u lg e d . M o tile . G r a m -p o sitiv e.
G e la tin s ta b : S tr a tifo r m li q u e f a c t io n .
D extrose agar s l a n t : M oderate, beaded, convex, g l i s t e n i n g , smooth,
tr a n sp a r e n t w h it e . Adherent to medium.
Litmus m ilk : S lo w ly becoming p ep to n ized in upper o n e - t h ir d , lower
p a r t s o f t curd w ith in d ic a t io n s o f gas p ro d u c tio n .
hydrogen s u l f i d e n e g a t iv e .
K oser1s c i t r a t e v a r ia b le .
S ta rch h y d ro ly ze d .
N i t r i t e s produced from n i t r a t e s .
In d o l n o t produced.
V oges-P roskauer v a r i a b l e .
A cid and gas produced from l a c t o s e , d e x tr o s e , s u c r o s e , m a lto s e , x y lo s e ,
1 -a r a b in o s e , r a f f i n o s e . I n e r t i n m a n n ito l.
P o ta to : Abundant, sp rea d in g , r a is e d , g l i s t e n i n g , d ir t y g ra y , medium
darkened.
A ero b ic, f a c u l t a t i v e .
A34
A e r o b a o illu s Group
Medium r o d s : 0 .3 to 0 . 6 micron i n w id th by 2 .8 to 3 .6 m icrons
lo n g , o ccu rr in g s in g ly and in p a ir s . Spores c e n t r a l, e lo n g a t e . Sporangia
s l i g h t l y b u lged to c y l i n d r i c a l . G r a m -p o sitiv e . M o tile .
G e la tin s t a b : S tr a tifo r m l i q u e f a c t io n .
D extrose agar s la n t : Abundant, beaded, con vex, g l i s t e n i n g , v e r r u o o se ,
tra n sp a r en t w h it e . Growth ad herent to medium.
Litmus m ilk : S o f t curd and red u ced . L ater becoming p ep to n ized in
upper h a l f o f tu b e , low er h a l f hard curd w ith ev id en ce o f gas fo rm a tio n .
Hydrogen s u l f i d e p ro d u ctio n v a r i a b l e .
K oser1s c i t r a t e p o s i t i v e .
S ta rch h y d r o ly z e d .
N i t r i t e s produced from n i t r a t e s .
In d o l n o t produced.
V oges-P roskauer v a r i a b l e .
A cid and gas from l a c t o s e , su c r o s e , d e x tr o s e , m a lto s e , x y lo s e ,
1 -a r a b in o s e , and r a f f i n o s e . No a c id from m a n n ito l.
P o ta to : Abundant, sp r e a d in g , co n v ex , g l i s t e n i n g , r u g o s e , gray.
Medium n o t changed. E vidence o f gas fo rm a tio n in bubbles a t s id e o f s l a n t .
A ero b ic, f a c u l t a t i v e .
29.
A36
B a o illu s adhaerens Group
S h ort r o d s : 0 .3 to 0 .7 m iorons w id e by C.8 to 1 .4 m iorons lo n g ,
o ccu rr in g s in g ly and in p a i r s . Spores c e n t r a l , sm all and round. Sporangia
c y l i n d r i c a l . G r a m -p o sitiv e. N o n -m o tile.
G e la tin s ta b : No li q u e f a c t io n .
D extrose agar s l a n t : M oderate, e o h in u la t e , r a is e d , g l i s t e n i n g ,
sm ooth, g r a y is h -w h ite . N on-adherent to medium.
Litmus m ilk : S lo w ly becom ing p e p to n iz e d .
Ifydrogen s u l f i d e n o t produced.
K oser1s c i t r a t e p o s i t i v e .
S tarch n o t h y d ro ly ze d .
N itr a te s reduced t o n i t r i t e s .
In d o l n o t produced.
V oges-Proskauer n e g a t iv e .
No a c id in c a rb o h y d ra tes.
P o ta to : M oderate, sp rea d in g , r a is e d , d u l l , r u g o se, p a le g r a y ish oran ge, medium darkened.
A ero b ic•
H a b ita t: F o r e -g u t.
B36
B a c illu s adhaerens Group
Large r o d s : 0 .8 4 to 1 .4 m icrons w id e by 2 .9 to 4 .3 m iorons lo n g ,
o c c u r r in g s i n g l y , i n p a ir s , and sh o r t c h a in s . Spores c e n t r a l , o v a l.
Sporangia c y l i n d r i c a l . G ra m -p o sitiv e. N o n -m o tile.
G e la tin s ta b : S tr a tifo r m li q u e f a c t io n .
Agar s l a n t : Abundant, sp rea d in g , r a i s e d , d u ll, r u g o s e , g ra y ,
ad h eren t to medium.
Litmus m ilk : S lo w ly becoming p e p to n iz e d .
Ifydrogen s u l f i d e p ro d u ctio n v a r ia b le .
K oser’ s c i t r a t e p o s i t i v e .
S ta rch h y d ro ly ze d .
N i t r i t e s produced from n i t r a t e s .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
Weak a c id p ro d u ctio n from d e x tr o s e , s u c r o s e , and m a n n ito l. No a c id in
m a lto s e , l a c t o s e , 1 -a r a b in o s e , x y lo s e , or r a f f in o s e .
P o ta to : Abundant, sp r e a d in g , r a i s e d , g l i s t e n i n g , sm ooth, d u ll o ran ge,
p o ta to darkened.
A ero b ic.
H a b ita t: F o r e -g u t.
30AS7
iia o illu B adhaeran Group
Medium r o d s i 0 .6 to 0 .8 m ioron w ide by 1 .4 to 2 .1 m icrons Ionjt,
o ccu rr in g s in g ly and in p a ir s . Spores lo n g er than b road, c e n t r a l .
Sporangia c y l i n d r i c a l . G r a m -p o sitiv e. Non-mo t i l e .
G e la tin s t a b i S tr a tifo r m li q u e f a c t io n .
D ex tro se agar s l a n t i Abundant, sp r e a d in g , r a is e d , d u l l , ru g o se,
d ir ty g r a y , n on -ad h eren t to medium.
Litmus m ilk : P e p to n iz a tio n commencing e a r ly and g o in g to com p letion
throughout th e tu b e .
Mydrogen s u l f i d e s tr o n g ly produced.
K o ser's c i t r a t e p o s i t i v e .
S ta rch h y d r o ly s is v a r ia b le .
N i t r i t e s produced from n i t r a t e s .
In d ol n o t produced.
V oges-Proskauer v a r i a b l e .
A cid from d e x tr o s e , s u c r o s e , and m a n n ito l. No a c id from m a lto se ,
l a c t o s e , x y l o s e , l- a r a b in o s e , or r a f f i n o s e .
P o ta to i Abundant, sp r e a d in g , r a is e d , g l i s t e n i n g , r u g o s e , g r a y is h brown, p o ta to darkened.
A erobic to s l i g h t l y f a c u l t a t i v e .
H ab itat t H in d -g u t.
B38
B a c illu s o ir o u la n s Group
R ods: 0 .5 to 0 .8 m icron wide by 2 .1 to 4 .2 m icrons lo n g , o ccu rrin g
s in g ly and in p a ir s . Spores term in al to s u b -te r m in a l, lo n g e r than broad.
Sporangia p l e o t r i d i a . G r a m -p o sitiv e. M o tile .
G e la tin s ta b : C rateriform l i q u e f a c t io n .
D extrose agar s l a n t : S ca n ty , e o h in u la t e , r a is e d , g l i s t e n i n g , smooth,
tra n sp a r en t g r a y , n on -ad h eren t to medium.
Litmus m ilk : S low ly becoming p e p to n iz e d .
Mydrogen s u l f i d e p ro d u ctio n s l i g h t .
K o ser's c i t r a t e n e g a t iv e .
S ta rch n o t h y d ro ly ze d .
N itr a te s n o t red u ced .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
A cid n o t produced from ca rb o h y d ra tes.
P o t a t o : S ca n ty , e o h in u la t e , r a is e d , d u l l , v e r r u c o sa , grayish -b row n ,
medium darkened.
A ero b ic.
H a b ita t: H in d -g u t.
-3 1 B39
Dao l H ub M isce lla n eo u s Group
Rodei 0 .3 to 0 .6 raioron w ide ty 1 .2 to 2 .8 m iorons lo n g , o ccu rrin g
s in g ly and i n p a ir s . Spores c e n t r a l , s n a i l , s l i g h t l y lo n g er than broad.
Sporangia s l i g h t l y b u lg e d . G r a m -p o sitiv e. M o tile .
G e la tin s ta b : Napiform to s t r a t if o r m li q u e f a c t io n .
D extrose agar s la n t* Abundant, sp r e a d in g , co n v ex , g l i s t e n i n g ,
sm ooth, cre a m -c o lo red , n on -adh eren t to medium.
Litmus m ilk : A lk a lin e .
Hydrogen s u l f i d e p ro d u ctio n v a r ia b le .
K oserf s c i t r a t e p o s i t i v e .
S ta rch n o t h y d ro ly zed .
N it r a t e s n o t red u ced .
In d o l produced.
V oges-P roskauer n e g a t iv e .
A cid produced from d e x tr o s e , 1 -a r a b in o s e , x y lo s e , and weakly from
m a n n ito l. No a c id produced from s u c r o s e , l a c t o s e , m a lto s e , or r a f f in o s e .
P otato* M oderate, sp rea d in g , r a i s e d , g l i s t e n i n g , r u g o s e , r e d d ish brown grow th, darkening o f medium.
A ero b ic.
MO
B a c illu s s u b t i l i s Group
Medium to la r g e rods * 0 .7 to 1 .1 m iorons w ide by 2 .4 to 4 .3 miorons
lo n g , o ccu rr in g s i n g l y , i n p a ir s , and i n sh o r t c h a in s . Spores c e n t r a l,
o v o id . Sporangia c y l i n d r i c a l . G r a m -p o sitiv e. M o tile .
G e la tin stab * S tr a tifo r m li q u e f a c t io n .
D extrose agar s la n t* Abundant, s p r e a d in g , r a is e d , r u g o s e , d ir t y g ra y , a d h eren t to medium.
Litmus m ilk* S lo w ly becoming p e p to n iz e d .
Hyi'rogen s u l f i d e p ro d u ctio n v a r ia b le .
K oserf s o i t r a t e p o s i t i v e .
Staroh s tr o n g ly h y d ro ly ze d .
N itr a te s reduced to n i t r i t e s .
In d o l n o t produced.
V oges-Proekauer n e g a t iv e .
Weak a o id p ro d u ctio n from m a n n ito l. No a o id produced from d e x tr o se ,
l a c t o s e , m a lto s e , s u c r o s e , x y lo s e , 1 -a r a b in o s e , or r a f f i n o s e .
P otato* Abundant, sp rea d in g , r a i s e d , g l i s t e n i n g becoming d u ll,
sm ooth, d u ll o ra n g e, medium darkened.
A erob ic.
H ab itat* F o r e -g u t.
32B41
H a o illu s s u b t i l i s Group
Large r o d s i 0*9 to 1 .1 mioron w id e by 1 .6 to 5 .7 m icrons lo n g ,
o ccu rr in g s in g ly , in p a ir s , and sh o r t c h a in s . Spores c e n t r a l , o v a l and
la r g e . Sporangia c y l i n d r i c a l . G r a m -p o sitiv e. M o tile .
G e la tin s ta b : S tr a tifo r m li q u e f a c t io n .
D extrose agar s la n t : Abundant, sp r e a d in g , r a is e d , g l i s t e n i n g ,
smooth to s l i g h t l y v e r r u o o se , g r a y is h -w h ite , n on -adherent to medium.
Litmus m ilk : S lo w ly becoming p ep to n ized w ith r e d u c tio n on th e bottom .
Hydrogen s u l f i d e produced.
K o ser's c i t r a t e p o s i t i v e .
S ta rch s l i g h t l y h y d ro ly ze d .
N i t r i t e s n o t produced from n i t r a t e s .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
No a c id from ca rb o h y d ra tes.
P o ta to : Abundant, sp rea d in g , r a i s e d , d u l l , smooth, d u ll orange,
medium darkened.
A ero b ic.
H a b ita t: H in d -g u t.
B42
B a c illu s s u b t i l i s Group
Large r o d s: 0 .7 t o 0 .8 m icron w ide by 1 .7 to 5 .7 m icrons lo n g ,
o ccu rr in g s i n g l y , i n p a ir s , and in sh o r t and lo n g c h a in s . Spores c e n t r a l
to s u b - c e n t r a l, o v a l. Sporangia c y l i n d r i c a l . G r a m -p o sitiv e. M o tile .
G e la tin s ta b : S tr a tifo r m li q u e f a c t io n .
D extrose agar s l a n t : Abundant, sp r e a d in g , con vex, g l i s t e n i n g ,
sm ooth, g ra y , n on -ad h eren t t o medium.
Litmus m ilk : S lo w ly becoming p e p to n iz e d .
Eydrogen s u l f i d e p roduced.
.
K oser’ s c i t r a t e p o s i t i v e .
S tarch h y d rolyzed s l i g h t l y .
N itr a te s n o t reduced to n i t r i t e s .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
No a c id from c a rb o h y d ra tes.
P o ta to : No grow th.
A ero b ic.
H a b it a t :
H in d - g u t •
—33
B43
B a c illu s e u b t i l i s Group
Large ro d s: 1 .4 to 1 .5 m icrons w ide by 4 .6 to 5 .0 m icrons lo n g ,
o ccu rr in g s in g ly , in p a ir s , and i n sh o r t and lo n g c h a in s . Spores c e n t r a l,
lo n g er than b ro a d . Sporangia c y l i n d r i c a l . G ra m -p o sitiv e. M o tile .
G e la tin s t a b : S tr a tifo r m l i q u e f a c t io n .
D extrose agar s la n t : Abundant, s p r e a d in g , convex, g l i s t e n i n g ,
smooth to v e r r u o o s e , g r a y is h -w h ite , n on -adh eren t to medium.
Litmus m ilk : S lo w ly becoming p ep to n ized above w ith r e d u c tio n b elo w .
Tfydrogen s u l f i d e p ro d u ctio n v a r ia b le .
K oser’ s c i t r a t e p o s i t i v e .
S tarch h y d r o ly s is v a r ia b le .
N itr a te s n o t red u ced .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
No a c id from ca rb o h y d ra tes.
P o t a t o : Abundant, sp rea d in g , r a i s e d , g l i s t e n i n g , v e r r u o o s e , d u ll
o ra n g e, medium darkened.
A e r o b ic .
Habita t : H in d -g u t•
B44
B a c illu s s u b t i l i s Group
Large r o d s : 0 .8 to 1 .6 m icrons w ide by 2 .2 to 4 .3 m icrons lo n g ,
o ccu rr in g s i n g l y , in p a ir s , and in lo n g c h a in s . Spores c e n t r a l ,
lo n g er then broad . Sporangia c y l i n d r i c a l . G ra m -p o sitiv e. M o tile .
G e la tin s ta b : S tr a tifo r m li q u e f a c t io n .
D extrose agar s l a n t : Abundant, s p r e a d in g , con vex, g l i s t e n i n g , smooth
to v e r r u o o s e , g r a y is h -w h ite , n on -ad h eren t to medium.
Litmus m ilk : S lo w ly becoming p ep to n ized w ith r e d u c tio n in bottom o f
tu b e .
Ifydrogen s u l f i d e p ro d u ctio n v a r ia b le .
K oser’ s c i t r a t e p o s i t i v e .
S ta rch s l i g h t l y h y d ro ly ze d .
N it r a t e s n o t red u ced .
In d o l n o t produced.
V oges-Proskauer n e g a t iv e .
No a c id from ca rb o h y d ra tes.
P o t a t o : Abundant, sp rea d in g , r a i s e d , d u l l , smooth, d ir t y oran ge.
A ero b ic.
Habit a t : Hind -g u t.
34A45
B a c illu s s u b t i l i s Group
Medium r o d s i 1 .2 to 1 .4 m iorons w id e by 2 .2 to 2 .9 m icrons lo n g ,
o ccu rr in g s in g ly and i n p a ir s . G r a m -p o sitiv e. M o tile . Spores c e n t r a l,
narrow , lo n g . Sporangia c y l i n d r i c a l .
G e la tin sta b s S tr a tifo r m li q u e f a c t io n .
D extrose agar s la n t s Abundant, sp rea d in g , r a i s e d , d u l l , ru g o se,
g r a y , ad h eren t to medium.
Litmus m ilk s Becoming p ep to n ized q u it e r a p id ly and f i n a l l y go in g
to co m p letio n throughout e n t ir e tu b e.
Ifydrogen s u l f i d e s t r o n g ly produced,
K osert S c i t r a t e p o s i t i v e .
S tarch s tr o n g ly h y d ro ly ze d .
N i t r i t e s produced from n i t r a t e s ,
In d o l n o t produced.
V oges-P roskauer v a r i a b l e .
Acid produced from d e x tr o se and s u c r o s e . Weak a c id p ro d u ctio n in
m a lto se and m a n n ito l. No a c id from l a c t o s e , x y lo s e , 1 -a r a b in o s e , or
r a ffin o s e .
P o t a t o s Abundant, sp r e a d in g , r a i s e d , d u l l , r u g o se , grayish -b row n ,
medium darkened.
A erob ic, f a c u l t a t i v e .
H a b ita ts H in d -g u t.
A46
B a c illu s s u b t i l i s Group
R odss 1 .4 to 1 .7 m iorons w ide by 3 .0 to 4 ,4 m iorons lo n g , occu rrin g
s i n g l y , i n p a ir s , and s h o r t c h a in s . Spores c e n t r a l , lo n g e r than broad.
Sporangia c y l i n d r i c a l . G ra n t-p o sitiv e. M o tile .
G e la tin sta b s S tr a tifo r m li q u e f a c t io n .
D ex tro se agar s la n t s Abundant, sp r e a d in g , f l a t , d u l l , s l i g h t l y
v e r r u o o s e , g ra y , n on -ad h eren t to medium.
Litmus m ilk s R a p id ly becoming red u ced i n bottom o f tu b e w ith fo rm a tio n
o f a s o f t cu rd . F in a lly p ep to n ized in upper tw o - th ir d s .
Ifydrogen s u l f i d e p o s i t i v e .
K osert s c i t r a t e p o s i t i v e .
S ta rch s tr o n g ly h y d ro ly ze d .
N i t r i t e s formed from n i t r a t e s .
In d o l n o t produced.
V oges-P roskauer n e g a t iv e .
A cid formed from d e x tr o s e , s u c r o s e , l a c t o s e , and m a lto s e . No a c id
i n m a n n ito l, x y lo s e , 1 -a r a b in o s e , or r a f f i n o s e .
P o ta to s Abundant, sp r e a d in g , r a is e d , d u l l , v e r r u o o se , crea m -co lo red ,
medium s l i g h t l y darkened.
A e r o b ic , f a c u l t a t i v e .
Habit a t s H in d -g u t•
35A47
B a o illu s s u b t i l l s Group
Large r o d s « 0 .8 to 1 .4 m icrons w ide by 2 .8 to 4 .3 m icrons lo n g ,
o ccu rr in g s in g ly and i n p a ir s . Spores la r g e and c e n t r a l , lo n g er than
b road . Sporangia c y l i n d r i c a l . G r a m -p o sitiv e. M o tile .
G e la tin stab* S tr a tifo r m li q u e f a c t io n .
D extrose agar s la n t* Abundant, sp rea d in g , r a is e d , d u l l , smooth,
g r a y , n on -ad h eren t to medium.
Litmus m ilk : P e p to n iz a tio n commencing e a r ly and go in g to com pletion
th rou gh -ou t tu b e .
Ifydrogen s u l f i d e produced.
K oser1S c i t r a t e p o s i t i v e .
S ta rch h y d r o ly s is s t r o n g ly p o s i t i v e .
N itr a te s reduced to n i t r i t e s .
In d o l n o t produced.
V oges-Pro skauer n e g a t iv e .
A cid from s u c r o s e , d e x tr o s e , and m a lto s e . No a c id i n l a c t o s e ,
m a n n ito l, x y l o s e , l- a r a b in o s e , or r a f f i n o s e .
P otato* Abundant, sp rea d in g , co n v ex , d u l l , v e r r u o o s e , g ra y , medium
n o t d is c o lo r e d .
A ero b ic, f a c u l t a t i v e .
H abitat* H in d -g u t.
848
B a c illu s s u b t i l l s Group
Medium r o d s: 0 .8 4 to 1 .1 m icrons w id e by 2 .2 to 2 .9 m icrons lo n g ,
o c c u r r in g s in g ly and in p a i r s . Spores c e n t r a l to s u b - c e n t r a l, lo n g er
than b road . Sporangia w ith rounded e n d s, v ery s l i g h t l y b u lg e d . Gramp o s i t i v e . M o tile .
G e la tin stab* S tr a tifo r m l i q u e f a c t io n .
D ex tro se agar s la n t* M oderate, e c h in u la t e , r a is e d , g l i s t e n i n g ,
sm ooth, c r e a m y w h it e , n on -adh eren t to medium.
Litmus m ilk* S lo w ly p e p to n iz e d .
Ifydrogen s u l f i d e p o s i t i v e ,
K oser’ s c i t r a t e p o s i t i v e .
S ta rch s tr o n g ly h y d r o ly z e d .
N i t r i t e s produced from n i t r a t e s .
In d o l n o t produced.
V oges-Proskauer v a r i a b l e .
A cid in d e x tr o s e , s u c r o s e , x y lo s e , and m a n n ito l. No a c id from
l a c t o s e , m a lto s e , l- a r a b in o s e , or r a f f i n o s e .
P otato* Abundant, sp r e a d in g , r a i s e d , d u l l , r u g o se, d ir t y gray,
medium darkened.
A erob ic.
H abitat* F o r e -g u t.
-3 6 A49
B a o illu s s u b t i l i s Group
Medium r o d s « 1 .1 to 1 .4 miorone w id e by 2 .6 to 2 .9 m iorons lo n g ,
o c c u r r in g s i n g l y , in p a ir s and in sh o r t and lo n g c h a in s . Spores c e n t r a l,
to s u b -c e n tr a l, lo n g e r than b road . Sporangia s l i g h t l y s w o lle n , ends
tr u n c a te . G r a m -p o sitiv e. M o tile .
G e la tin s t a b : S tr a tifo r m li q u e f a c t io n .
D extrose agar s la n t : Abundant, sp r e a d in g , co n v ex , g l i s t e n i n g ,
v e r r u o o s e , tr a n s p a r e n t w h it e , n on -ad h eren t to medium.
Litmus m ilk : S lo w ly becoming p ep to n ized in upper o n e - t h ir d , reduced
i n low er p o r tio n .
Ttydrogen s u l f i d e s l i g h t l y produced.
K oser1s c i t r a t e p o s i t i v e .
S tarch h y d ro ly ze d .
N itr a te s reduced to n i t r i t e s .
In d o l n o t produced.
V oges-Proekauer n e g a t iv e .
A cid i n d e x tr o s e , s u c r o s e , and m a n n ito l. No a c id i n l a c t o s e , x y lo s e ,
m a lto s e , 1 -a r a b in o s e , or r a f f i n o s e .
P o ta to : Abundant, sp rea d in g , r a i s e d , g l i s t e n i n g , sm ooth, creamyg r a y , medium darkened.
A ero b ic.
B60
B a c illu s s u b t i l i s Group
Large r o d s: 1 .1 to 1 .4 m icrons w id e by 2 .8 6 to 4 .3 m icrons lo n g ,
o c c u r r in g s i n g l y , i n p a ir s and i n sh o r t and Ionf c h a in s . Spores c e n t r a l,
o v o id , la r g e . Sporangia c y l i n d r i c a l . G r a m -p o sitiv e. M o t ile .
G e la tin s t a b i S tr a tifo r m li q u e f a c t io n .
D extrose agar s l a n t : Abundant, sp r e a d in g , con vex, g l i s t e n i n g ,
sm ooth, g r a y is h -w h ite , non -adh eren t to medium.
Litmus m ilk : S lo w ly becoming p ep to n ized w ith r e d u c tio n i n the
bottom .
Ttydrogen s u l f i d e produced,
K oser1s c i t r a t e p o s it iv e *
S ta rch h y d r o ly s is s l i g h t to n e g a t iv e .
N i t r i t e s produced from n i t r a t e s .
In d o l n o t produced.
V oges-Proskauer n e g a t iv e .
A cid from d e x tr o s e , s u c r o s e . No a c id produced from l a c t o s e , m a lto s e ,
m a n n ito l, x y lo s e , 1 -a r a b in o s e , or r a f f i n o s e .
P o ta to : Abundant3 sp rea d in g , r a i s e d , g l i s t e n i n g , sm ooth, d u ll o ra n g e,
medium darkened s l i g h t l y .
A ero b ic.
H a b ita t: H in d -g u t.
-3 7
TABLE I
J
Il
t + i si
I
i
* ♦ I V
- t + %Z
:
- Z + I *
I
(
*
- I _ * .
- t - * -
-
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I
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89 t
t
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t
t
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B12*
Al 4
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t
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:
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t
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*
*
*
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+
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I
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t
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I
I +
*
* V
i
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I
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*
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*
*
*
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♦
Z
-
:
-
i
I Z
-
Z
•
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S
-
J
I
:
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:
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B
BR
: + t A : - * Z
8
Z
8
2A 8 - 8 I + *
* - * -
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:
S
A
:
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SIs i A
ip ep /iO
BR
I V *
8
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I
8
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I
+
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I
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8
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8
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S
♦
I
+
-
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M
* V t 2
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2
8 A 8 - 2
* - * - *
Sfierocoocus
M icrococcus Spe
M icrococcus Spe
Z
+
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(GRA'^-POSITIVE)
Litmus
Milk
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to
COCCI
+
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ARC
+
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V
8
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8
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8
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8
8
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A *
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A 8 - 8- I - I
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- 8
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2 A
S
2
8
8
ic r o c o c c u s su b fla v u s
ic r o c o c c u s ep id erm id is
ic r o c o c c u s ep id erm id is
ic r o c o c c u s ep id erm id is
M icrococcus Spe
M icrococcus S p e
M icrococcus
Spe
M icrococcus
M icrococcus
Spe
Spe
ic r o c o c c u s
Spe
ic r o c o c c u s
Spe
ic r o c o c c u s
Spe
2
8
! —
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'ic r o c o c c u s fr e u d e n r e ic h ii
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S S
SI
CO S
A l?
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Al 8 t $
B19 t
B20 i
B21 S *
B22 I B23 I A24
A25
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A28 « ♦ t + i 2
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A30 t ♦ I + S +
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8
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A32 2 - 8 — 2 -
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: pep I - < A *
i
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tpep/ftC S ♦ I A Z
*pep/RC I ♦ Z A I
8
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$
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A I 2
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2
8
A z
A
Bacterium
Bacterium
Bacterium
Bacterium
acteriu m
sp*
sp*
sp*
sp*
sp*
8
Bacterium sp*
Hacterium sp*
2
8
8 A z
A Z
A I A :
2
2
8
2 ■ 8 - 8
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Bacterium sp*
Z
Bacterium sp*
Bacterium sp*
B act rium sp*
Bacterium sp*
•35'
TABLE I I I
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*
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3
C
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:s l ; i
I
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43
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5
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+
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B59
MO
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M2
MS
B44
A45
A46
A47
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A49
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8
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s i :
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11
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pep/RCG 2 + IAG *AG tAG tAG t - tAG tAG tAG
*
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2
t
t
t
t
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2
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*
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2
*
t
t
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P0 p 2 + $ A 2 - 2 A 2 - 2 A 2 - t - 2 2
8 1
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x
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pap i T t A t - t A t - t A i - 1 - 1 .
2
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t
t
t
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1
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t
8 ®
3 •8 C
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! *! ■ I 2 Litrm
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* A er o b a e illu s
* Asro ^aci i r ts
* ""
* _B* Ad-'acro-s
I
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1
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1 2»
*
2 B.
1 3.
: B.
* B.
*
I*
t
+
POP
RA/pep
pop
pop
pep/R
t
A
A
s u b tilis
s u b tilis
S U b t ilis
subtiIj g
s u b t i Ii &
: B. su’itilis
: B. subtilis
2 B. subtilis
s 3 . subtilis
subtilis
t Be subtilis
Grp.
Grp.
Grp.
G rp .
Grp.
Grp.
r .
Grp.
rp.
rp.
Grp.
Grp.
Grp.
Grp.
Grp.
-4 0 DISCUSSION
!h ere i s an o b v io u s la c k o f g ram -n egative sh o r t rods in th e b a c t e r ia l
f l o r a o f th e a lim en ta ry t r a c t o f G r y llo b la tta oam podeiform is as compared
w ith th e f l o r a o f o th er i n s e c t s as found by S tein h a u s ( 1 9 4 1 ) .
Over
60 per c e n t o f th e b a c t e r i a l f l o r a o f th e in s e c t s examined by t h a t worker
was made up o f g ra m -n eg a tiv e sh o r t r o d s .
S tein h a u s s t a t e d th a t he had
g iv e n th e p e r c e n ta g e s m erely fo r th e purpose o f making approxim ate
com parisons and t h a t he d id n o t in te n d t h a t t h i s data sh o u ld be in te r p r e te d
as th e a c tu a l p erce n ta g e s o f th e v a r io u s groups o f b a c t e r ia found in i n s e c t s
in g e n e r a l.
A p o s s ib le e x p la n a tio n o f g ra m -n eg a tiv e organism s in th e g r y l l o b l a t t i d
i s to be found in th e e c o lo g y o f th e i n s e c t , s in c e in r e s p e c t to h a b ita t
i t d i f f e r s so w id e ly from o th e r i n s e c t s .
S tein h a u s (1941) r e p o r te d no g ra m -n eg a tiv e sh o r t rods from any o f th e
s p e c ie s he s tu d ie d o f th e ord ers Odonata, Homopt e r a , or IKmenopte r a .
However from th e order O rthoptera ( t o w hich G r y llo b la tta oam podeiform is
b e lo n g s) he i s o l a t e d g ra m -n eg a tiv e sh o r t rods from 4 s p e c ie s o f th e o r d er,
nam ely, B l a t t e l l a germ anloa L ., Neombius f a s o i a t u s , v a r . f a s c ia t u s DeG.,
Conocephalus f a s o i a t u s , v a r . f a s c ia t u s DeG., and Diapheromera fe n o r a ta , Say.
Of th e 60 s t r a in s o f b a c te r ia in c lu d e d in t h i s pap er, IG (32 per c e n t)
were c o c c i o f th e gen u s M icrococcu s, 16 (32 per cent) were g r a m -p o sitiv e
asporogenous rods o f th e genus B acterium , and 18 (36 per c e n t ) were gram­
p o s i t i v e sporogenous rod s o f th e genus B a c i l l u s .
The m o rp h o lo g ic a lly d i s t i n c t s e r i e s o f organism s found in t h is
in v e s t ig a t i o n appear to f a l l n a t u r a lly in t o a s e r ie s o f c l o s e l y r e la t e d
-4 1 grou p s•
The members o f t h e s e groups may d i f f e r s l i g h t l y from one an oth er
b u t th ey appear more c l o s e l y r e la t e d both m o r p h o lo g ic a lly and p h y s io lo g ic a lly
to each o th e r than th ey do to members o f any o th e r grou p .
The v a rio u s
members o f each group w hich d i f f e r to v a r y in g degrees w ith regard to
morphology and p h y s io lo g ic a l r e a c t io n s e r e p robably s t r a in s o f the seme
s p e c ie s .
Of th e o o c c i th e r e appear to be 11 groups o f s t r a i n s , a l l o f which
b elo n g to th e genus M icrocooou s.
S tr a in s A6, AS, and A7 appear to be
s l i g h t v a r ia n t s o f M icrococcus e p id e n a id is ( K lig le r ) Huoker.
S tr a in A4
appears to be a v a r ia n t o f M iorooooous su b fla v u s Bumm, s t r a in AlS a
v a r ia n t o f M icroooocus fr e u d e n r e io h ii G u ille b e a u .
None o f th e o th er
s t r a in s o f th e M ioroooooi co u ld be c l a s s i f i e d to s p e c ie s .
The 16 s t r a in s o f g r a m -p o s itiv e asporogenous rods appear to f a l l
in t o 9 gro u p s.
A ll o f t h e s e are p la c e d in th e genus Bacterium sin o e th ey
d id n o t correspon d w ith th e d e s c r ip t io n o f any o th er g en u s, and th ey d id
n o t s e r io u s ly v i o l a t e th e g e n e r ic d e s c r ip t io n c f B acterium , which i s n o t,
how ever, a sh a rp ly d e fin e d grou p .
Of 18 s t r a in s o f th e genus B a c illu s th e r e appear to be 8 groups o f
s tr a in s .
S tr a in s A33 and A54 s r e members o f th e A e r o b a o illu s Group.
S tr a in s A55, B3f>, and AS? a r e members o f th e B a o illu s adhaerens Group.
AS? i s p o s s ib ly a v a r ia n t o f B a o illu s p a n is M igula.
B38 b e lo n g s to th e
B a o illu s e ir o u la n s Group and i s p o s s ib ly a v a r ia n t o f B a c illu s s e r u s it id u s
La C o ste .
1339 b elo n g s to th e M isce lla n eo u s Group.
B40, B41, B42, B43, and
B44 are probably s t r a in s o f a s in g le s p e c ie s o f th e B a o illu s s u b t i l l s Group,
A45, A46, A47, B48, A49 and B60 are s t r a in s o f another s p e c ie s o f the
>-42->
!.Iacillu s s u b t i l i s Group.
S tr a in s A l? , A l? , 319, 321, and A27 w ere very pleom orphic forn$
w hich a cco rd in g to some a u th ors m ight bo d e sig n a te d as c o o o o b n o illu s .
In
some o f th e s e c u lt u r e s th e r e appeared la r g e , p ear-sh ap ed organism s and
r o d s , w h ile In o th er c u lt u r e s th e r e appeared rods and c o c c i .
As th e c u lt u r e s
r e ta in e d t h i s c h a r a c t e r is t ic a f t e r b ein g r e p la te d f i v e tim e s i t was d ecided
to be pltiomerphism ra th o r th an co n ta m in a tio n .
D iese c u lt u r e s were p la ced
in th e genus Bacterium s in c e th e r e was a preponderance o f d e f i n i t e rods in
a l l in s t a n c e s .
Ifo s i g n i f i c a n c e can be a tta c h e d to th e f a c t th a t th e f l o r a o f th e two
specim ens o f i n s e c t s in d ic a t e d by 1A1 s t r a in s and ' B' s t r a in s o f organism s
d i f f e r a p p r e c ia b ly s in c e , a s p r e v io u s ly s t a t e d , no attem p t was made to
determ ine w hether th e f l o r a o f th e i n s e c t was c o n s ta n t.
R egarding th e d is t r ib u t io n o f th e organism s in th e a lim en ta ry t r a c t ,
a la r g e m a jo r ity o f th e c o c c i and n o n -sp o r u la tin g rods were found in th e
a n te r io r p o r tio n o f th e t r a c t , w hereas a m a jo rity o f th e e p o r u la tin g rods
were found i n th e p o s te r io r p o r t io n .
Ihe 50 s t r a in s r e f e r r e d to r e p r e s e n t a much sm a lle r number o f s p e c ie s .
The number o f s p e c ie s , w ould , no doubt, vary w ith th e in d iv id u a l making th e
in t e r p r e t a t io n depending on how l i b e r a l or c o n s e r v a tiv e he in te r p r e te d th e
d ata p r e s e n te d .
Perhaps th e most i n t e r e s t i n g o b se r v a tio n made in t h is
in v e s t ig a t i o n i s
th a t so many d if f e r e n t k in d s o f organism s can be i s o l a t e d
from two in d iv id u a ls o f a s in g le s p e c ie s o f i n s e c t , so few o f which can be
i d e n t i f i e d as p r e v io u s ly d esc r ib e d s p e c i e s .
Other in v e s t ig a t o r s may w e ll
tak e is s u e o f t h i s r e p o r t t h a t 50 s t r a in s o f organism s can be i s o la t e d from
-4 3 aiqr suoh s o u r c e , so few o f w hich can be d e f i n i t e l y c l a s s i f i e d to s p e c ie s
i n Bergey*s Manual.
The f i f t h and l a t e s t e d it io n o f B ergeyt B Manual
c o n ta in s d e s c r ip t io n s o f 1335 s p e c ie s w ith a r e fe r e n c e to th e o r ig in a l
p la c e o f p u b lic a t io n o f 5 ,6 0 0 d e s c r ip t io n s .
Probably no b a o te r io lo . 1 s t
w i l l q u e s tio n th e o p in io n t h a t th e r e a r e v ery numerous s p e c ie s o f
u n d escrib ed b a c t e r ia .
As has been p r e v io u s ly s t a t e d in v e s t ig a t i o n o f th e
b a c t e r i a l f l o r a o f i n s e c t s a re few , c l a s s i f i c a t i o n o f organism s found even
fe w e r .
The v a r io u s s c a t t e r e d r e p o r ts r eg a r d in g th e b a c t e r ia l f lo r a o f th e
a lim en ta ry t r a c t s o f i n s e c t s e x h ib it a d e f i n i t e la c k o f a ttem p ts to
a c c u r a te ly id e n t if y th e organism s tu d ie d , w ith th e e x c e p tio n s o f S tein h au s
(1 9 4 0 ),
ifhite (1 9 0 6 ), and v ery few o t h e r s .
in c lu d e many h it h e r t o u n d e so r ib ed .
E v id en tly th e s p e c ie s found
D escrib ed s p e c ie s o f b a c t e r ia i s o la t e d
from i n s e c t s are co m p a ra tiv ely few in B ergey1s Manual.
L i t t l e i s known o f th e fo o d h a b it s o f G r y llo b la tta oam podeiform is.
M ills and Pepper (1 9 3 7 ) s t a t e , "GryI l o b l a t t i d s which were k e p t fo r
o b s e r v a tio n were fe d a d i e t o f f l i e s and w h ite bread .
They a ls o fed on
decaying o r g a n ic m atter g le a n e d from th e moss or mulch in w hich th ey were
k e p t.
The f l i e s w hich th ey were fe d were k i l l e d b e fo r e b ein g p la c e d w ith
them .
L ive or even s l i g h t l y moving f l i e s e x c it e d th e g r y l l o b l a t t i d s
g r e a t l y , and th ey oou ld n o t be induced to ta k e o th er than co m p letely
m o tio n le s s p r e y .”
I f d ecayin g o rg a n ic m atter i s a p a rt o f th e d ie t o f th e
i n s e c t , or i f i t con tam in ates t h e ir fo o d , one would e x p e c t a more v a r ie d
b a c t e r i a l f l o r a in t h i s i n s e c t than in phytophagous and ju ic e -s u c k in g
in s e c t s .
Ttie author does n o t in te n d t h a t t h is paper s h a ll be in t e r p r e t e d a s a
-4 4 com p lete stud y o f th e b a c t e r ia l f lo r a o f G r y llo b la tta oampod p iform !s .
TO
accom p lish such a stu d y a la r g e r number o f specim ens o f th e i n s e c t would
have to be exam ined.
On th e b a s is o f 50 s t r a in s o f organism s from two
specim ens o f th e i n s e c t one can r e a d ily s e e how th e number o f s t r a in s o f
organism s would in c r e a s e w ith each a d d it io n a l specim en exam ined.
This
would in c r e a s e th e scop e o f th e problem to such an e x t e n t t h a t i t would be
a f u l l tim e problem fo r a g r e a te r p erio d o f tim e than one norm ally spends
a c q u ir in g a m asters d eg ree .
An attem p t would a ls o have to b e made to
o b ta in a medium more s u it a b le fo r th e growth req u irem en ts o f many o f th e
s t r a in s which d ie a f t e r a few weeks on d e x tr o se agar though t r a n s f e r s are
fr e q u e n tly made.
-46'
SUMMARY
1.
F i f t y s t r a in s o f b a c t e r ia were i s o l a t e d from th e a lim en tary
t r a c t o f th e o rth o p tera n G r y llo b la tta oam podeiform ls campod elfo rm !s
W alker.
2.
M orp h ological and p h y s io lo g ic a l s t u d ie s were made o f th e s e 60
s t r a in s w ith an attem p t t o i d e n t i f y each o f them.
3.
Of th e 50 s t r a i n s , 16 (32 per c e n t ) were g r a m -p o s itiv e c o c c i o f th e
genus M icrococcu s, 16 (32 per c e n t) were g r a m -p o sitiv e asporogenous rods o f
th e genus Bacterium , and 18 (36 per c e n t) were g r a m -p o sitiv e sporogenous r o d s .
!IVro o f th e s e l a t t e r s t r a in s were o f th e A e r o b a o illu s Group, th r e e o f th e B a c illu s
adhaerens Group, one o f th e B a o lllu s c ir o u la n s Group, one o f th e Mis c e lla n e o u s
Group, and th e rem aining 11 s t r a in s com prised two s p e c ie s o f th e B a c illu s
s u b t i l i s Group.
4.
Other in t e r p r e t a t io n s may be made o f th e d ata in c lu d e d in t h is
paper r e s u lt i n g in a somewhat d if f e r e n t grouping o f th e organism s s tu d ie d .
The p h y s io lo g ic a l c h a r a c t e r is t ic s upon w hich one would la y m ost s t r e s s i s
an im p ortan t f a c t o r in d eterm inin g w hether 2 v a ry in g s t r a in s sh ou ld be
grouped to g e th e r or n o t .
How much a s t r a i n may vary from a d escrib ed s p e c ie s
and s t i l l bo c l a s s i f i e d as t h a t s p e c ie s i s a v e r y im p ortan t f a c t o r a l s o .
5.
W hile th e fin d in g o f p r e v io u s ly u n desorib ed s p e c ie s o f b a c te r ia
in th e i n t e s t i n a l t r a c t s o f in s e c t s i s to be a n t ic ip a t e d , ju d g in g from
p rev io u s in v e s t i g a t i o n s , th e number o f such s p e c ie s i s o l a t e d from
G r y llo b la tta oampod elfo rm is was found to be e s p e c ia l ly h ig h .
40LITBRATURE CITED
S erg ey , DeHe e t a l (1939) Sergey* s Manual o f D eterm in a tiv e B a cter io lo g y #
6 th ed# Hie W illiam s and W ilk in s Co#, B a ltim o re. 1032 pp.
Crampton, O.C. (1916) The Thoraoio S o l e r i t e s and th e S y ste m a tic P o s it io n
o f O r y llo b la t t a oam podelform ie W alker, A Remarkable Arm eotant,
" O r th o p te r o id " I n s e c t . B n t. News 2 0 , 337-360
F orb es, S .A . (1 8 8 2 ) Bacterium a P a r a s it e o f th e Chinch Bug.
1 6 , 824-826
Amer, N a t .,
Glasgow, Hugh (1 9 1 4 ) The G a stric Caeca and th e C aeoal B a c te r ia o f th e
H e te r o p te r a . B io l . B u ll. £6 101-170
Gurney, A .B . (1 9 3 7 ) S y n o p sis o f The G r y llo b la t t id a e With The D e sc r ip tio n
o f A New S p e c ie s From Oregon. P a n -P a c ific E n to m o lo g ist,
13 , No. 4 1 5 9 -1 7 1 .
L ey d ig , F.
(1 8 5 7 ) Lehrbuoh der H is t o lo g ie dee Mensohen und der Thie r e .
F ran k fu rt A. M. p .3 3 7 : As quoted by Glasgow i n above r e fe r e n c e .
M i l l s , H.B. and Pepper, J .H . (1987) O b servation s on G r y llo b la tta
oam podeiform is W alker. Ann. E n t. S oo. Amer. 3 0 , 269-274
P i e r c e , (1921) S a n ita r y Entom ology.
B oston .
R ichard G. Badger.
The Gorham P r e s s .
S tein h a u s , E.A . (1940) The M icro b io lo g y o f I n s e c t s w ith S p e c ia l R eferen ce
to th e B io lo g ic R e la tio n s h ip s Between B a c te r ia and I n s e c t s .
Baot R eview s. 4 , 17-67
S te in h a u s, E .A . (1 9 4 1 ) The B a c te r ia l F lo ra o f th e A lim en tary T racts o f
T h irty s p e c ie s o f I n s e c t s . 54 pp. Tb be p u b lis h e d . (Seen in
m an u scrip t)
Walker, E.M. (1 9 1 4 ) A new s p e c ie s o f O rthoptera form ing a new genus and
fa m ily . Can. E n t. 4 6 , 93-99
W hite, G.F. (1 9 0 6 ) The b a c t e r ia o f th e a p ia ry w ith s p e c ia l r e fe r e n c e to
b ee d is e a s e s . U .S . D ept. A g r ., Bur. E n t., Tech. B i l l . , Ho. 14,
50 pp.
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PAGES 57 - 58 - 59
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Aoid r e a c tio n
13
- A lk a lin e r e a c tio n
C - - - - - - - - - - - - - - - - - - - - - - - - -
- Curd
G - . . . - - - - - - - - - - - - - - - - - - - - - I - . - . - .
- - - - - - - - - - - - - - - - - - -
- - - - - - - - - - - - - - - - - - - - - - - - - - -
Gas form ation
- in e r t
n e g a t iv e
p o s itiv e
p6p
----- - - - ----- - - p e p to n iz a tio n
R - - - - . - - - - - - - - - - - - - - - - - - - - s i ------------------------- - - - - - - - - - - - - - -
Reduced
-------- - s l i g h t
V - - . - - - - - - - - - - - - - - - - - - - - - - -
v a r ia b le
d a ta f o llo w in g / s i g n i f i e s r e a c tio n in
low er p o r tio n o f tu b e