STUDIES OH m
FTIOLOGY OF
SPHOIFIC INFHOTIOTTg BOVINE

PYHLOWRPITIS
T h e s is f o r t h e D eg ree o f P h .D .
M ich ig an S t a t e C o l l e g e
E rn est S ta r F a e n stra

19^7
An A b s t r a c t
P art I .

P a t h o g e n i c i t y o f C o ry n e b a c te rlu m r e n a l e f o r R a b b i t s .

I n o r d e r t o d e te r m in e t h e e f f i c a c y o f t h e r a b b i t a s a n e x p e r i m e n t a l
a n im a l f o r u s e i n t h e s t u d y o f b o v in e p y e l o n e p h r i t i s ,

w h i t e r a b b i t s w ore

i n o c u l a t e d i n t r a v e n o u s l y w i t h a s u s p e n s i o n o f C o r y n e b a c t e r 1urn r e n a l e w h ic h
h a d b e e n grown on tr y n to s © a g a r ( D i f c o ) a t 37° C f o r ?U h o u r s .

T h e se

r a b b i t s w ere necfcop sied a t i n t e r v a l s o f o n e - h a l f h o u r , o n e , tw o, t h r e e ,
fo u r, fiv e , six ,
in o c u la tio n .

seven, n in e , e le v e n , e ig h te e n and n in e te e n days a f t e r

H em oglobin d e t e r m i n a t i o n s , t o t a l l e u k o c y t e c o u n t s and

d i f f e r e n t i a l l e u k o c y t e c o u n t s w e re made on a l t e r n a t e d a y s .
b lo o d , lu n g s, l i v e r ,

s p l e e n , k i d n e y , s t e r n a l b o n e marrow and u r i n e w ere

c u l t u r e d on t r y p t o s e a g a r .
liv e r,

The h e a r t

F o r m i c r o s c o p i c s t u d y , s p e c im e n s o f t h e lu n g ,

s p l e e n , k i d n e y , u r e t e r s and u r i n a r y b l a d d e r w ere f i x e d i n Z e n k e r 's

f l u i d ; n a r a f f i n s e c t i o n s p r e p a r e d , a nd s t a i n e d w i t h h e m a t o x y l i n and e o s i n
a n d b y t h e Q ram -W eigert m ethod f o r s t a i n i n g b a c t e r i a i n s e c t i o n s .
The e a r l i e s t change i n t h e k id n e y was t h e a c c u m u l a t i o n o f n s e u d o e o s i n o p h i l s i n t h e c a p i l l a r i e s a nd c o l l e c t i n g t u b u l e s w i t h c o n g e s t i o n i n
t h e p a p i l l a r y r e g i o n o f t h e m e d u lla on t h e s e c o n d day.

F o llo w in g t h i s , a t

t h r e e a nd f o u r d a y s a f t e r i n o c u l a t i o n , n e c r o s i s and e r o s i o n o f t h e p a p i l l a
w e re fo u n d .

M asses o f b a c t e r i a w ere found i n t h e r e m a in s o f c a p i l l a r i e s

and tu b u le s and i n th e d e b r is i n th e p e l v i s .

The p o ly m o r p h o n u c le a r

l e u k o c y t e ( p s e u d o e o s i n o p h i l ) was t h e p re d o m in a n t in f l a m m a to r y c e l l fo u n d i n

th e e a rly le sio n .

On a h o u t t h e f o u r t h day t h e r e n a l p e l v i c m ucosa a p p e a r e d

n e c r o t i c and th e p p r i p e l v i c t i s s u e s c o n ta in e d s e ro u s f l u i d , f i b r i n , e r y t h r o ­
c y t e s and p s e u d o e o s i n o p h i l s .

Lym phocytes a p p e a r e d on t h e s i x t h day and

m o n o n u a le a r p h a g o c y t e s and a c t i v e f i b r o b l a s t s a p p e a r e d on t h e s e v e n t h d ay .
T h e s e l a t t e r t h r e e c e l l s i n c r e a s e d i n number a s tim e p a s s e d w h i l e t h e p s e u d o e o s i n o p h i l s d e c r e a s e d i n number b u t n e v e r d i s a p p e a r e d e n t i r e l y .

At 17 a n d

IS d a y s t h e r e w ere c h an g e s I n t h e c o r t e x s u c h a s a n i n c r e a s e i n i n t e r s t i t i a l
c o n n e c tiv e t i s s u e ,
o f ly m p h o c y te g .

d i l a t i o n and c o n s t r i c t i o n o f t u b u l e s a n d a c c u m u l a t i o n s

The b l o o d p i c t u r e was one c h a r a c t e r i s t i c o f a c u t e i n f e c ­

t i o n s and showed c h a n g e s su c h a s a n i n c r e a s e i n l e u c o c y t e s on t h e f o u r t h
day a n d an i n c r e a s e i n p e r c e n t o f p s e u d o e o s i n o p h i l s w i t h a d e c r e a s e I n
p e r c e n t o f ly m p h o c y te s i n t h e d i f f e r e n t i a l l e u k o c y t e c o u n t .
P art II.

P r o p e r t i e s o f Some C o l o n i a l P h a s e V a r i a n t s o f C o r y n e b a c te r lu m
re n a le .

P iv e c u l t u r e s , r e p r e s e n t i n g a t l e a s t t h r e e d i f f e r e n t c o lo n y t y p e s
w ere c h o s e n from s t o c k c u l t u r e s o f C o ry n e b a c te rlu m r e n a l e f o r s tu d y i n o r d e r
t o d e te r m in e t h e i r m o r p h o l o g i c a l , b i o c h e m i c a l , s e r o l o g i c a l and p a t h o g e n i c
r e l a t i o n sh ip s.
One c u l t u r e w hich m igh t b e c l a s s i f i e d a s " sm o o th 11 showed t h e w e a k e s t
a c t i o n i n th e fe rm e n ta tio n o f g lu c o se , d i g e s t i o n o f c a s e in ,

showed t h e l e a s t

r e s i s t a n c e t o t h e b a c t e r i c i d a l a c t i o n o f b o v i n e p la s m a , a n d was t h e l e a s t
v iru le n t fo r ra b b its.
Two c u l t u r e s w hich m ig h t b e ' c l a s s i f i e d a s " ro u g h " showed t h e m ost
r e s i s t a n c e t o t h e b a c t e r i c i d a l a c t i o n o f b o v i n e p la s m a a n d w ere t h e most
v iru le n t fo r ra b b its.
The o t h e r two c u l t u r e s w ere I n t e r m e d i a t e I n rao rp h o lo ey a n d p a t h o g e n i ­
c ity .
The r e s u l t s o f a g g l u t i n a t i o n t e s t s showed t h a t t h e r e w ere a n t l a e n i c

d i f f e r e n c e s a n d s i m i l a r i t i e s b e tw e e n t h e d i f f e r e n t c u l t u r e s h u t t h e r e was
l i t t l e o r no c o r r e l a t i o n w i t h t h e m o r p h o l o g i c a l c h a r a c t e r i s t i c s o f t h e
c o lo n ie s.

M a jo r P r o f e a

STUDIES ON THE ETIOLOGY OF SPECIFIC
INFECTIOUS BOVINE PYELONEPHRITIS

"by
E rn e s t S t a r F e a n g tr a

A Thesi s
Submitted, to th e School of G raduate S tu d ie s o f Michigan
S t a t e C ollege of A g r i c u l t u r e and A pp lied S c ie n c e
i n p a r t i a l f u l f i l l m e n t of t h e re q u ire m e n ts
f o r th e degree of
DOCTOR OF PHILOSOPHY

Deoartment of Animal P a th o lo g y
19U7

ACKNOWLSDG-EMENTS

The w r i t e r hereby acknowledges th e a i d extended
by Dr. Frank Thorp, J r . , a s major p r o f e s s o r and Dr. E. T.
Hallman as hea.d of th e departm ent o f Animal P a th o lo g y .
Also v e ry h e l p f u l was th e a d v ic e of Dr. I . F,
Huddleson i n suggesting; a fundam ental annroach to th e
stu d y of th e m icroorganism , th e a d v ic e of Dr. R. F.
Langham i n p h o to g ra p h ic te c h n i c , and th e t e c h n i c a l
a s s i s t a n c e of ?ir. M. L. Gray i n making hemoglobin
d e te r m in a tio n s and le u c o c y te c o u n ts .
The c o o p e r a tio n and a d v ic e of the guidance
committee composed o f Drs. F. Thorp, J r . , E. T.
Hallman, H. J . S t a f 3e th , W. L. Kallmsnr. and f . A.
H opp°rt were v e ry much a pp reciated..

Table of C o n te n ts

1

In tro d u c tio n
P art I .

P a t h o g e n i c i t y o f C orvnebacterium r e n a l e f o r
ra b b its.
Review of t h e L i t e r a t u r e

2

M a te r ia ls and Methods

2

R e s u lts

U

D is c u s s io n

2U

SumiTiary

28

R efe re n c e s

J>0

Illu stra tio n s
P art I I .

P r o p e r t i e s o f Some C o lo n ia l Phase V a r ia n ts
of Corynebacteriurn r e n a l e .
In tro d u ctio n

56

A g g lu tin a tio n R e l a t i o n s h i p

57

S u s c e p t i b i l i t y to Bovine Plasm a

5$

B iochem ical P r o p e r t i e s

60

P a t h o c e n i c i t y f o r R a b b its

6l

D is c u s s io n

68

Summary

69

R eferen c es

70

Illu stra tio n s

71

INTRODUCTION
F o r many y e a r s l a b o r a t o r y a n im a ls have been
c o n s id e r e d r e f r a c t o r y t o i n f e c t i o n w ith Corynebacterlum
re n a le .

The a d v a n ta g e s of th e u s e o f th e sm all la b o r a ­

to r y a n im a ls i n t h e stu d y of d i s e a s e i s obvio us.

A

f o r t u i t o u s c irc u m sta n c e I n t h i s l a b o r a t o r y p o i n t e d t o
fo n f,

V

i o o a 6 ? r \ o r [

+ rs

\\o

i n f e c t i o n under c e r t a i n c o n d i t i o n s .

2 M 2 C S D ^ 1 1 } ! .S

^0

The f i r s t p a r t of

t h i s t h e s i s i s d e v o ted to a s tu d y o f th e p a t h o g e n i c i t y
of C. r e n a l e i n th e r a b b i t .
The second p a r t of t h i s t h e s i s i s de v o ted t o
a s tu d y o f t h e co lo n y morphology o f c u l t u r e s o f
£• r e n a l e and t h e r e l a t i o n o f t h e s e m o rp ho log ical
c h a r a c t e r i s t i c s to th e p a th o g e n ic and a n t i g e n i c
p ro p e rtie s.

P a rt I .

PATHOGPNICITY OF COIFHEBACTER ITJM RENALE FOR RABBITS

The ad v an tag es of t h e use of th e sm all l a b o r a t o r y anim als i n th e
stu d y of th e d is e a s e s of la r g e anim als a r e w ell known.

This i s p a r t i c u ­

l a r l y t r u e i f a s i m i l a r d is e a s e p r o c e s s i s ev inced by th e la b o r a t o r y
P re v io u s to 19 U6 , la b o r a t o r y an im als were c o n s id e re d t o be

an im als.

r e f r a c t o r y t o i n f e c t i o n by Corynebacterlum r e n a l e .
F n d e rle n (1891) r e p o r te d t y p i c a l r e n a l l e s i o n s e ig h t and e le v e n
days a f t e r in tr a v e n o u s i n o c u l a t i o n of an organism i s o l a t e d from a c a s e
of bovine n y e l o n e p h r i t i s i n t o two r a b b i t s w ith l i g a t e d u r e t e r s .
was re p e a te d by L o ll ( l^ L ? ) .

This

Jones and L i t t l e ( 1936 ) r e p o r t e d nega.tive

r e s u l t s a f t e r u s in g w h ite mice, guinea pirns and r a b b i t s .

L o v e ll ( 19^ 6 )

and L ovell and Cotchin ( 19^ 6 ) d e s c rib e e x p e rim e n ta l p y e l o n e p h r i t i s i n
mice which r e s u l t e d from t h e in tr a v e n o u s i n o c u l a t i o n of 20 m i l l i o n
organisms.

The r e s u l t s a f t e r in tr a v e n o u s i n o c u l a t i o n o f 60 m i l l i o n

organisms i n t o guinea p ig s and r a b b i t s th e y r e p o r t p.g much l e s s s a t i s ­
fa c to ry .
onto

They a l s o d e s c r ib e th e r e s u l t s of i n o c u l a t i o n of C. r e n a l e

th e c h o r i o - a l l a n t o i c membrane of n in e day o ld c h ic k embryos.
'’’he p a th o g e n i c ity of C. r e n a le f o r th e r a b b i t was d is c o v e re d

in

t h i s l a b o r a t o r y w hile a tte m p tin g to p rod u ce immune s e r a i n r a b b i t s f o r
s e v e r a l c o l o n i a l phase v a r i a n t s of C. r e n a l e .

A t o t a l of nine r a b b i t s

d ie d of th e 21 s t a r t e d on immune serum p r o d u c tio n .

E xperim ents were

then designed, to study the p a th o g e n e s is of e x p e rim e n ta l p y e l o n e p h r i t i s
in ra b b its.
M a te r ia ls and Methods
F i f t e e n r a b b i t s (a b o u t 2 Kg each) were in o c u l a t e d in tr a v e n o u s l y
w ith a p p ro x im a te ly f o u r b i l l i o n m icroorganism s each.

The

c u ltu re

( C u ltu re No. 2 , T h e s is , P a r t I I ) chosen f o r i n o c u l a t i o n was from a cow

-

3-

w ith a t y p i c a l case of p y e l o n e p h r i t i s ; i t s a t i s f i e d a l l m orphological#
c u l t u r a l and b io c h em ica l re q u ire m e n ts f o r C. r e n a l e (Bergey, 1939 ) s-n(l
had p roved p a th o g e n ic f o r s e v e r a l r a b b i t s .

The f i r s t t e n r a b b i t s were

n e c ro p s ie d o n e - h a lf ho u r, one, two, t h r e e , f o u r , f i v e ,
and eleven days a f t e r i n o c u l a t i o n .

s i x , seven, n in e

The rem ain in g f i v e were allo w ed t o

l i v e f o r 18-19 days i n o rd e r t o determ in e t h e m o r t a l i t y which p r e v a i l e d
under th e c o n d itio n s of th e experim ent (one d ie d a t e i g h t d a y s).

Blood

samples were ta k en on a l t e r n a t e days t o a llo w d e te r m in a tio n of any
changes i n hemoglobin l e v e l s (C enco-Sheard-Sanford p h o te lo m e te r , Hoffman
method, 19U l), numbers of le u k o c y te s , and d i f f e r e n t i a l le u c o c y te c o un ts

(100 c e l l s c o u nted on each s l i d e ; b r i g h t s t a i n ) .

In most c a s e s t h e h e a r t

b lo o d , lu n g s, l i v e r , s p le e n , kidney, s t e r n a l bone marrow and u r in e were
c u l t u r e d on tr y p t o a e a g a r ( D if c o ) .

T is s u e smears of lu n g , l i v e r , s p le e n ,

k id n e y , u r i n e and bone marrow were made and s ta i n e d w ith gram s t a i n .
Gross p a t h o l o g i c a l changes were re c o rd e d .

F o r m ic ro s c o p ic s tu d y , s p e c i­

mens of th e lu n g , l i v e r , s p le e n , k id n e y , u r e t e r s and u r i n a r y b la d d e r
were f i x e d i n Z e n k e r's f l u i d ; p a r a f f i n s e c t i o n s p r e p a r e d , and s t a i n e d
w ith h em atoxylin and e o sin and by th e Gram-Weigert method f o r s t a i n i n g
b a c te ria in sec tio n s.
A f t e r th e f i r s t experim ent was com pleted a n o th e r was designed to
i n v e s t i g a t e more th o ro u g h ly th e happenings o f th e f i r s t , f o u r days.

F ig h t

r a b b i t s ( s e r i e s A) were u sed , one was k i l l e d a s a normal c o n t r o l a t t h e
tim e th e o t h e r seven were in o c u la te d and th e s e were n e c r o p s ie d a t th e
r a t e o f two a t th e end o f one day, two days, and th r e e days, and one a t
th e end of f o u r days.

The t e c h n i c s and p ro c e d u re s were s i m i l a r to th o s e

employed i n t h e f i r s t exp erim ent.
Almost w ith o u t e x c e p tio n th e l i v e r s e x h i b i t e d l e s i o n s of
c o cc id io sis.

These l e s i o n s were re c o rd e d b u t w i l l not be r e f e r r e d to i n

-li­

t h e i n d i v i d u a l case r e p o r t s .

This c o n d itio n may e x p la i n th e high

i n i t i a l le u k o c y te co un ts o b ta in e d i n some c a s e s , e s p e c i a l l y i n th e
se rie s A ra b b its .
O c c a s io n a lly , th e l e s i o n s r e p o r t e d i n th e g ro s s exam ination
were m issed i n s e c t i o n i n g th e t i s s u e .

However, s u f f i c i e n t l e s i o n s were

found to a llo w adeq uate d e s c r i p t i o n s of th e m icroscop ic changes.
R e s u lts
Case r e p o r t s of r a b b i t s used i n th e experim ent fo llo w .

The

r e s u l t s of t h e study of t h e t i s s u e smears so n e a r ly p a r a l l e l e d th e
r e s u l t s o f t h e c u l t u r e work t h a t th e y w i l l not be f u r t h e r r e p o r te d .
Case 1
R a b b it 1 was n e c r o p s ie d o n e - h a lf ho u r a f t e r i n o c u l a t i o n .
Organisms were c u l t u r e d from th e h e a r t b lo o d , l i v e r , lu n e and k id n e y .
No g ro ss l e s i o n s were found.
M icroscopic f i n d i n g s .

D ip h th e ro id s were found i n th e l a r g e r

b lo o d v e s s e l s of th e l i v e r and lung.

The s e c t i o n of t h e lun g showed

c o n g e s tio n , s l i g h t se ro u s e x u d a tio n , and i n th e a l v e o l a r c a p i l l a r i e s
were many n s e u d o e o s in o p h ils , some of which appeared t o c o n ta in
d ip h th e r o id s .
Time

Hb

*TBC

M

L

P

B

s

0

13-3

8 ,1 00

1

75

19

u

1

13-3

8 ,8 50

2

73

2U

1

0

1 /2 h r .

(See next page f o r key to h e a d in g s)

5-

-

Time - I n t e r v a l a f t e r I n o c u la tio n ; 0 “being j u s t p r i o r to
in o c u l a t i o n .
Kb

- Hemoglobin i n grams p e r 100 ml blood.

WBC

- Leucocytes p e r cubic mm.

M

- Monocytes, p e r c e n t .

L

- Lymphocytes, p e r c e n t.

P

- Polym orphonuclear le u k o c y te s (p s e u d o e o s in o p h ils ) p e r c e n t .

B

- B a s o p h ils , p e r c e n t .

E

- E o s in o p h ils , p e r c e n t.
Case 2

R abb it 2 was n e c r o p s ie d one day a f t e r i n o c u l a t i o n .
Ciub'uTHU. ifOm tii« ll'v e ? y.iiu. kidney*

Organisms

ffG v i s i u l e l e e i o n a were Oreeexi't

ex cep t a p o s s i b l e enlargement, o f th e s p le e n .
M icroscopic f i n d i n g s .

Time

Hb

0

13.9
11.8

1 day

The s p le e n was co n g este d .

WBC

M

L

P

B

E

10,900

1

12

2U

3

0

8,800

3

H6

U9

2

0

Case 3
R ab b it 3 was n e c r o p s ie d two days a f t e r i n o c u l a t i o n .
were c u l t u r e d .

No organism s

The s p le e n a p p eared s l i g h t l y sw ollen.

M icroscopic f i n d i n g s .

A s l i g h t c o n g e s tio n o f th e sp le e n , e x ce ss

mucus i n t h e pulmonary b r o n c h i, and one fo c u s (250 raicra d ia m .) i n t h e
o u te r medulla of th e k id ney c o n s i s t i n g of p s e u d o e o s in o p h ils i n t h e t h i c k
lim bs o f H e n l e '8 loops were th e only a p p a r e n t l e s i o n s .
Time

Eb

WBC

o

io . i

8 ,9 0 0

2

9 .0

U.650

M

l

L

79
5

63

P

is
30

B

E

2

o

2

0

-6 -

Ca se U
R ab b it U was n e c r o p s ie d t h r e e days a f t e r I n o c u l a t i o n .
c u l t u r e d from t h e kidney and u r i n e .

Organisms were

The s p le e n seemed s l i g h t l y e n la r g e d .

The k id n e y s were co n g este d and t h e r e were s e v e r a l lo n g , s le n d e r , r a d i a t i n g
( toward th e c o r te x ) a b s c e s s e s i n th e m edulla.
M icroscopic f i n d i n g s .
a n ce.

The l i v e r was somewhat foamy in a p p e a r­

There were s e v e r a l f o c i ( 0 . 5-1 ™ i n d ia m .) i n th e r e n a l m edu lla

c o n s i s t i n g m ainly of p s e u d o e o s in o p h ils and n e c r o t i c t i s s u e d e b r i s .

There

were a l 90 many sm all l e s i o n s i n th e m ed u lla which a p p ea re d to be th e
b e g in n in g s o f t h e above mentioned f o c i c o n s i s t i n g of n s e u d o e o s in o n h ils i n
c a p i l l a r i e s and tu b u le s (m ainly t h i c k lim bs and c o l l e c t i n g tu b u le s ) and
c o n g e s tio n of th e c a p i l l a r i e s i n th e a r e a .

There were e r y t h r o c y t e s ,

p s e u d o e o s in o p h ils and t i s s u e d e b r is i n th e p e l v i s .

The n n r t s of th e t i n s

o f th e p a p i l l a e found i n t h e s t a i n e d s e c t i o n s showed s l i g h t n e c r o s i s .
Time

Hb

WBC

M

L

0

11. U

8.550

1

73

9-5

g, 200

6

6o

1 0 .g

15.900

5

57

?. days

3

"

B

E

1

1

2g

6

0

37

1

0

P

Case 5
R ab b it 5 was n e c r o p s ie d fo u r days a f t e r i n o c u l a t i o n .
were c u l t u r e d from t h e k id n e y , u r i n e and h e a r t blo od .
sw ollen and c o n g e ste d .

Organisms

The kid neys were

There was p a p i l l a r y n e c r o s is w ith r a d i a t i n g

l e s i o n s e x te n d in g thro u gh th e medulla tow ard th e c o r te x .

The u r i n e was

b lo o d tin g e d .
M icroscopic f i n d i n g s .

The s p le e n e x h i b i t e d an i n c r e a s e i n pseudo­

e o s in o p h i ls and much p h a g o c y to s is o f b lo o d pigment by th e p h a g o cy te s.

The

u r e t e r s were e n la rg e d , th e e p ith e liu m was n e c r o t i c and desquamated, h y a l i n e

t o d i e s were p r e s e n t I n t h e lam ina p r o p r i a , and th e lumens were f i l l e d
w ith e r y t h r o c y t e s , p s e u d o e o s in o p h ils , f i b r i n , d ip h th e r o id s and t i s s u e
d e b r is .

A ll th e s t r u c t u r e s i n t h e r e n a l p a p i l l a e i n an a r e a about 1 mm

t h i c k were n e c r o t i c .

Masses of b a c t e r i a were p r e s e n t i n what had been

tu b u le s and c a p i l l a r i e s .

P s e u d o e o s in o p h ils were s c a t t e r e d th rou gh o ut th e

l e s i o n and a p p ea re d to c o n t a i n d i p h t h e r o i d s .

S e v e ra l c o l l e c t i n g t u b u l e s

and c a p i l l a r i e s i n th e in n e r medulla were f i l l e d w ith p s e u d o e o s in o p h ils
and c e l l u l a r d e b r i s .

There was a n e c r o t i c fo c u s i n th e p e l v i c mucosa

w ith th e p r e s e n c e of s e r o f i b r i n o u s e x u d a te , c o n g e s tio n , hemorrhage and
p s e u d o e o s in o p h ils .

There was a s l i g h t amount of cloudy s w e llin g of th e

oroxim al co n v o lu te d t u b u l e s .
L

p

B

E

1

69

30

0

0

6 ,6oo

5

55

32

8

0

25.500

13

39

Ul

7

0

Time

Hb

'TBC

M

0

11.8

5,300

2 days

10.1

U H

11.8

Case 6
R ab b it 6 was n e c r o p s ie d f i v e days a f t » r i n o c u l a t i o n .
was a p re g n a n t fem ale which d ie d s h o r t l y b e fo re n ecro p sy .

T his anim al

Organisms were

c u ltu r e d from th e l i v e r , lun g, sp le e n , h e a r t b loo d, bone marrow, k id n e y ,
p l a c e n t a , f e t a l f l u i d and f e t u s .
sp le e n was s l i g h t l y e n la r g e d .
g e s te d .

The lu ng showed a few hemorrhages.

The

The k id n e y s were much sw o llen and con­

The kidney p e lv e s were e n la rg e d and f i l l e d w ith bloody f l u i d .

The p a p i l l a e were n e c r o t i c w ith r a d i a t i n g a b s c e s s e s i n th e m edulla.
were a ls o s e v e r a l s u b c a p s u la r a b s c e s s e s .
M icroscopic f i n d i n g s .

The u r e t e r s were e n la rg e d .

There were v a c u o le s i n th e h e p a ti c c e l l s

i n c e n t r a l p a r t of th e l o b u le s s u g g e s tiv e of f a t t y R e g e n e ra tio n .
lungs were s l i g h t l y co n g este d .

There

The

The s p le e n was c o n g e ste d and t h e r e was

-.2-

p h a g o c y to s is of e r y t h r o c y t e s end 'blood p ig m en t.

The u r e t e r s were d i s ­

tended. w ith "blood, t i s s u e d e h rig , and " b ac te ria .

Much o f th e u r e t e r a l

e p ith e liu m was n e c r o t i c and th e lam ina p r o p r i a was i n f i l t r a t e d w ith
p s e u d o e o s in o p h ils .

The r e n a l p a p i l l a e were eroded,

l a r g e masses of

b a c t e r i a and c e l l u l a r d e b r i s were observed i n th e c o l l e c t i n g tu b u le s i n
t h e medulla, around th e n e c r o t i c a r e a .

The p e l v i c e p ith e liu m was n e c r o t i c

and th e n e r i p e l v i c c o n n e c tiv e t i s s u e c o n ta in e d e r y t h r o c y t e s , f l u i d ,
f i b r i n , and p s e u d o e o s in o p h ils .

Some oro x im al co nvoluted tu b u le s showed

a 9l i f h t amount of cloudy s w e llin g .
Time

Hb

WBC

M

0

12.9

5.350

2 days

10.2

9.8

U

"

L

P

B

E

2

58

30

2

2

5,H oo

7

1+5

^7

1

0

11,050

12

3H 1+7

1

0

Case 7
R abbit 7 was n e c r o p s ie d s i x days a f t e r i n o c u l a t i o n .
were c u l t u r e d from the k id n e y and u r i n e .
e n la r g e d .

Organisms

The s p le e n ap p ea re d s l i g h t l y

The l e f t k id n e y showed e r o s io n o f p a p i l l a and c o r t i c a l and

s u b c a p s u la r a b s c e s s e s .

The p a p i l l a of th e r i g h t kidney was normal,

a lth o u g h th e c o r t e x c o n ta in e d a l a r g e w h ite l e s i o n which was cone shaped
w ith th e apex i n th e m edulla.

The u r e t e r s were s l i g h t l y c o n gested,

e s p e c i a l l y n e a r th e k id n e y s .
M icroscop ic f i n d i n g s .

The l i v e r was s l i g h t l y co n g este d , th e

c e n t r a l v e in s were en larg ed and t h e r e was a sm all (2 mm d ia m .) a b s c e s s
n e a r th e s u r f a c e .

The lumens of th e u r e t e r s c o n ta in e d b a c t e r i a .

The

r e n a l m edullae c o n ta in e d s c a t t e r e d f o c i which c o n s is te d o f groups o f
p s e u d o e o s in o p h ils and c e l l u l a r d e b r is and which were surrounded by an a r e a
of c a p i l l a r y c o n g e s tio n .

The t i p of th e l e f t r e n a l p a p i l l a was l o s t from

t h e s t a i n e d s e c t i o n s hut th e p e l v i c mucosa o p p o s ite th e p a p i l l a e x h i b i t e d
a n e c r o t i c fo c u s w ith i n f i l t r a t i o n of th e a r e a by s e ro u s and f i b r i n o u s
e x u d ate s, p s e u d o e o s in o p h ils and a few lymphocytes.

Time

Hb

WBC

M

L

P

B

E

0

13*3

8 ,9 00

1

6U

33

2

0

2 days

11.1

7.350

5

62

29

H

0

U

"

12.1

17,000

lU

52

33

1

0

6

"

12.7

1U.700

5

Ul

U8

5

1

Ca39 8
R ab b it 8 was n e c r o p s ie d seven days a f t e r i n o c u l a t i o n .
,.i

„

«*

v.

a

a
ilJL
IU
S
X I
1
4
L
U
.

a

i l l

W
Ui l l

i*

1

i U
1

were c u l t u r e d from th e k idn ey .

~

.3 .

U
f
c
l^
y» M
U
.L
L W
tl3

V m v. A
U
iU
i lU
U
liU
*

This
A^» —4 ^

M

V
i e,d
,u
*D
I
U
O

The s p le e n app eared s l i g h t l y e n la r g e d .

The kid neys were much e n la r g e d , e x h i b i t i n g p a p i l l a r y n e c r o s i s w ith p u ru ­
l e n t m a t e r i a l i n th e p e lv e s , and c o r t i c a l and s u b c a p s u la r a b s c e s s e s .
u r e t e r s were e n la rg e d and con gested.

The

The b la d d e r was f i l l e d w ith b loody

u rin e .
M icroscopic f i n d i n g s .

The l i v e r c e l l s showed cloudy s w e llin g and

th e r e were v a c u o le s i n d i c a t i n g c e n t r a l f a t t y d e g e n e ra tio n .

The pulmonary

a l v e o l a r w a l l s were th ic k e n e d by an i n c r e a s e of p s e u d o e o s in o p h ils and
e r y th r o c y te s .
b lo o d pigm ent.

The s p le e n was co n g este d and showed some p h a g o c y to s is o f
The u r i n a r y b la d d e r e p ith e liu m app eared s l i g h t l y d e g en e r­

a t e d and th e lumen c o n ta in e d b a c t e r i a and t i s s u e d e b r i s .

The u r e t e r s

were e n la rg e d and th e e p ith e liu m was n e c r o t i c ; b a c t e r i a and t i s s u e d e b r i s
were found i n th e lumens.

An ab sc e ss (1 x 1.5 nun) i n th e r e n a l m edulla

n e a r th e p a p i l l a r y t i p c o n ta in e d p s e u d o e o s in o p h ils and t i s s u e d e b r i s w ith
b a c t e r i a on th e b o rd e r.

There was a zone o f n e c r o s is a lo n g th e t i p o f

th e r e n a l p a p i l l a about .25 mra t o 1 .5 mm i n th ic k n e s s surrou nd ed by c o n g e s t!

-1 0 and an i n f i l t r a t i o n w ith inflam m atory c o l l 3.

S e v e ra l c o r t i c a l a b s c e s s e s

woro p r e s e n t and some of th e c o n v o lu ted t u b u le s e x h i b i t e d the c h a r a c t e r ­
i s t i c s of cloudy s w e llin g .

D ip h th e ro id s and p s e u d o e o s in o p h ils were

s c a t t e r e d through th e t i s s u e d e b r is i n th e n e l v i s .

The p e l v i c mucosa

was n e c r o t i c and th e o s r i p e l v i c t i s s u e c o n ta in e d a s e r o f i b r i n o u s ex u d ate ,
p s e u d o e o s in o p h ils , lymphocytes, mononuclear p h ag ocy tes and a c t i v e f i b r o ­
S e v e ra l a r e a s (b -1 0 mm diam. ) o f t u b u l a r d e g e n e ra tio n and

b la sts.

hemorrhage in th e c o r t i c e s and m edullas appeared to be hemorrhagic
in fa rc ts.
Hb

WBC

M

L

p

B

E

0

12.9

6,850

0

27

12

1

0

2 days

11 . S

7, Hoc

0

36

39

18

0

H

"

11. 8

10.U50

7

50

28

lH

1

6

"

11.8

15,600

21

32

U5

2

0

7

"

. 10.0

i\U 5 0

25

23

Uq

3

0

Time

Ca se 9
R abb it 9 was n e c r o p s ie d nine days a f t e r i n o c u l a t i o n .
were c u l t u r e d from th e kid ney and u r i n e .
e n la r g e d .

Organisms

The s p le e n appeared s l i g h t l y

The kidneys were v e ry s l i g h t l y e n la rg e d and e x h ib ite d p a p i l l a r y

n e c r o s i s and p y e l i t i s ( y e llo w is h l e s i o n 0.5 cm in d ia m e te r ).

The r i g h t

kidney a l s o had a sm all w hite c o r t i c a l l e s i o n .
M icroscopi c t'iod.lngs.

The pulmonary a l v e o l a r w a lls were s l i g h t l y

congested and c o n ta in e d an in c r e a s e d number of p s e u d o e o s in o p h ils .

The

s p le e n was c o n g e ste d and e x h i b i t e d a m oderate p h a g o c y to s is o f b lo o d
pigment.

The u r e t e r a l lumens c o n ta in e d some t i s s u e d e b r i s and the e p i­

t h e l i a l c e l l s c o n ta in e d a few h y a lin e b o d ie s (c y to p la s m ic ) .

The k id n e y s

were co n g este d and showed th e c h a r a c t e r i s t i c p a p i l l a r y e ro s io n ( p r o g r e s s i v e
n e c r o s is , masses of b a c t e r i a , i n f i l t r a . t i o n w ith p s e u d o e o s in o p h ils , and

-1 1 -

c o n g e s tio n ) .

The p e l v i s was f i l l e d w ith "blood, t i s s u e d e b r i s , b a c t e r i a

and p s e u d o e o s in o p h ils : t h e p e l v i c mucosa was n s c r o t i c and th e p e r i p e l v i c
t i s s u e s were c o n g e ste d and c o n ta in e d f i b r i n , lymphocytes, mononuclear
p hago cytes and p r o l i f e r a t i n g f i b r o b l a s t s .

There were lym phocytic f o c i

s c a t t e r e d th ro u g h o u t th e c o r t e x and p e r i p e l v i c t i s s u e .

The c o n v o lu te d

tu b u le s were s l i g h t l y d e g e n e ra te d a s ev id en ced by t h e p re s e n c e of cloudy
s w e llin g .
Time

Hb

TOC

M

L

P

B

S

0

12.3

5,600

0

73

22

5

0

2 days

10. u

8,250

0

69

25

5

1

U

"

11.1

9.250

3

72

If?

7

0

6

■

11. 8

12.350

7

60

28

5

0

9

"

12.U

16,550

1

3?

55

12

0

Case 10
R abbit 10 was n e c r o p s ie d e le v e n days a f t e r i n o c u l a t i o n .
were c u l t u r e d from th e k idn ey and u r i n e .

Organisms

The kid n ey s and u r e t e r s seemed

to be of about average s i z e and p r e s e n t e d no e x te r n a l l e s i o n s .

The l e f t

k id n e y showed p a p i l l a r y n e c r o s i s w ith p u r u l e n t m a t e r i a l i n th e p e l v i s .
M icro sco pic f i n d i n g s .
t i o n of th e a l v e o l a r w a l l s .

The lu n g s e c t i o n showed a s l i g h t conges­
There was a s l i g h t i n c r e a s e i n th e number of

p s e u d o e o s in o p h ils i n th e s p le e n .

The u r i n a r y b la d d e r e p i t h e l i a l c e l l s

c o n ta in e d a few h y a lin e b o d ie s ( c y to p la s m ic ) .

A few s c a t t e r e d c o l l e c t i n g

tu b u le s of b o th kidneys were f i l l e d w ith c e l l u l a r d e b r i s , a ls o b oth
c o r t i c e s e x h i b i t e d some cloudy s w e llin g and f a t t y d e g e n e re tio n .

There

was th e c h a r a c t e r i s t i c p a p i l l a r y e r o s io n of th e l e f t k id n e y w ith e a r l y
fib ro b la st p ro life ra tio n .

Also p r e s e n t was marked n e c r o s i s o f th e p e l v i c

mucosa and i n t h e p e r i p e l v i c t i s s u e were found in flam m ato ry c e l l s such a s

-1 2 -

pa e u d o e o s in o p h ils , lym phocytes, mononuclear p h a g o cy te s and p r o l i f e r a t i n g
fib ro b la sts.
L

p

B

E

0

69

29

2

0

8,950

k

67

26

3

0
0

Time

Hb

WBC

M

0

1 1 .g

7 . 75°

2 days

11.1

k

"

10.1

10,900

g

51

CVJ

9

6

"

11.6

12.550

H

53

3U

9

0

9

"

11.3

i3,Uoo

H

50

U6

0

0

11

"

10. g

9,1*50

3

33

56

g

0

A
u>-i,y a

M » OOI

Case 11
T> . n-v M ±

sxs. u u i u

______ * __
•11
>i _Wits
.u o u i__' u___u__s_ii i«e a
j

-

»c?v<Su vSoIl

-

Organisms were c u l t u r e d from th e kidney and u r i n e .
r a t h e r e m aciated .

» 4 ~
X 11'

The c a r c a s s was

The k id n e y s were e n la r g e d and d is p la y e d l a r g e p a l e

a r e a s i n th e c o r t i c e s , p a p i l l a r y n e c r o s i s and e r o s io n , and n e c r o t i c
d e b r is i n th e p e lv e s .

The l e f t u r e t e r was s l i g h t l y e n la r g e d .

M icroscopic f i n d i n g s .
c y t o s i s of b lo o d pigm ent.

The s p le e n was c o n g e ste d and showed phago­

One o f th e u r e t e r s e x h i b i t e d n e c r o s i s and

desquamation of th e e p ith e liu m .

The mucosa was cong ested and i n f i l t r a t e d

w ith lymphocytes w h ile th e lumen c o n ta in e d b a c t e r i a , p s e u d o e o s in o p h ils
and t i s s u e d e b r i s .

The r e n a l c o r t i c e s showed a s l i g h t i n c r e a s e in

a c t i v i t y o f i n t e r s t i t i a l c o n n e c tiv e t i s 3ue, c o n s t r i c t i o n o f masses of
tu b u le s and d i l a t i o n of o t h e r s , n o ta b ly th e c o l l e c t i n g tu b u le s , d i s t a l
co nv o luted tu b u le s and some few pro xim al co n v o lu te d t u b u l e s .
were co n g este d .

The m edullae

The p a p i l l a e were erod ed and th e l e s i o n s were surrou nd ed

by a zone of c h ro n ic inflam m atory p r o c e s s e s .
p s e u d o e o s in o p h ils , b a c t e r i a and t i s s u e d e b r i s .

The p e lv e s were f i l l e d w ith
The p e l v i c e p ith e liu m was

n e c r o t i c and th e p e r i p e l v i c t i s s u e s c o n ta i n e d sero u s f l u i d , p s e u d o e o s in o p h ils ,

-

13-

lym phocytes, mononuclear p h ag ocy tes and p r o l i f e r a t i n g f i b r o b l a - t s .

There

were sm all f o c i of lymphocytes s c a t t e r e d throughout th e k id n e y s.
Time

Hb

WBC

M

L

p

B

s

0

12.3

5,Hoo

2

50

Hi

6

1

2 days

11.6

7.590

9

H7

37

7

0

H "

ll.l

28,650

9

51

33

7

0

6

h

12.3

7.850

11

31

50

6

2

9

n

13-3

52,200

15

38

U6

1

0

11

«

13.3

19,950

15

37

Hi

7

0

lH "

12.9

16,200

12

Hi

Hi

6

0

17

13.5

9,Uoo

13

39

H6

l

1

»

.

Case 12
R ab b it 12 was n e c ro p s ie d sev e n te en days a f t e r i n o c u l a t i o n .
organisms were c u l t u r e d from any of th e org a n s.

There appeared to be

organisms i n th e s ta in e d smears o f th e k id n e y and u r i n e .
i n good p h y s i c a l c o n d itio n .

Wo

The c a r c a s s was

The k id n ey s were of a v erag e s i z e and e x h i b i t e d

p y e l i t i s , -p a p illa ry n e c r o s i s and a c c u m u la tio n of n e c r o t i c d e b r is i n th e
p e lv e s .
M icro sco pic f i n d i n g s .

The pulmonary a l v e o l a r w a lls c o n ta in e d a

s l i g h t l y i n c r e a s e d number o f p s e u d o e o s in o p h ils .

The u r e t e r a l e p ith e liu m

was n e c r o t i c and th e w a lls were th ic k e n e d w ith edematous f l u i d , conges~
t i o n , p s e u d o e o s in o p h ils , lymphocytes and mononuclear p h a g o c y te s .

The

k idneys e x h i b i t e d a m oderate amount of p a p i l l a r y n e c r o s i s (250 m icra
t h i c k ) and t h e c e l l s of th e c o l l e c t i n g t u b u l e s i n th e a r e a s c o n ta in e d
h y a lin e b o d ie s .

The v e ry t i p s of th e p a p i l l a e were not i n th e s e c t i o n s .

The c o r t i c e s e x h i b i t e d a s l i g h t in c r e a s e i n i n t e r s t i t i a l c o n n ec tiv e t i s s u e ,
d i l a t i o n of some t u b u le s and c o n s t r i c t i o n o f o th e r s .

Some t u b u le s showed

-l»levidance o f d e g e n e ra tio n and a tro p h y .
s c a t t e r e d th ro u g h o u t th e k id n e y s.
p h i l s and t i s s u e d e b r is .

A few lym phocytic f o c i were

The p e lv e s c o n ta in e d p seu do eo sin o ­

The p e l v i c mucosa was n e c r o t i c i n some a r e a s

and h y p e r p l a s t i c i n o t h e r s , and th e p e r i p e l v i c t i s s u e c o n ta in e d in c r e a s e d
numbers of c o n n e c tiv e t i s s u e elem en ts, lymphocytes and mononuclear phago­
c y te s .
M

I

p

B

E

10,150

0

79

16

5

0

13.1

11,000

u

65

2k

7

0

61

22

12

1

Time

Hb

0

12.9

2 days

WBC

u

n

11.1

2^,550

6

ft

12.3

iU ,050

3

62

2U

10

1

9

It

13.8

15,650

2

70

25

3

0

11

It

12.5

25.UOO

6

57

32

5

0

lU

II

1U.6

10,000

9

U5

32

lU

0

17

n

19.0

17.750

5

35

52

7

1

Case 15
R ab bit 13 was n e c r o p s ie d e ig h te e n days a f t e r i n o c u l a t i o n .
isms were c u l t u r e d from th e kidney and th e u r i n e .
c o n d itio n .

Oraan-

The c a r c a s s was i n p oor

The kid neys were s l i g h t l y e n la rg e d and e x h ib ite d w h ite c o r t i ­

c a l l e s i o n s c a u s in g th e s u r f a c e to appear m o ttle d , m e d u lla r a b s c e s s e s ,
p a p i l l a r y e r o s io n , p y e l i t i s and a cc u m u la tio n of p u r u l e n t f l u i d i n th e
p e lv e s .
M icro sco pic f i n d i n g s .

The r e n a l c o r t i c e s e x h i b i t e d an i n c r e a s e

i n i n t e r s t i t i a l c o n n ec tiv e t i s s u e , a r e a s of d i l a t i o n and of c o n s t r i c t i o n ,
and a l s o a tro p h y and d e g e n e ra tio n of some of th e t u b u l e s .

One m edulla

c o n ta in e d an a b sc e ss c h a r a c t e r i z e d by t i s s u e d e b r is , p s e u d o e o s in o p h ils
and mononuclear p h a g o cy te s.

S e v e ra l sm a ll f o c i s i m i l a r to th e above

a b sc e ss were a l s o p r e s e n t .

There was d i l a t i o n o f th e p a p i l l a r y d u c ts

and s l i g h t p a p i l l a r y e r o s io n ; th e dominant inflam m atory c e l l b e in g th e
mononuclear p h ag ocy te.

There was much n e c r o s i s of th e p e l v i c mucosa.

The p e l v i s c o n ta in e d t i s s u e d e b r is and " b a c te ria , and i n th e p e r i p e l v i c
t i s s u e p s e u d o e o s in o p h ils , lymphocytes, mononuclear p h a g o cy te s and p ro ­
l i f e r a t i n g f i b r o b l a s t s were p r e s e n t .
L

p

B

E

0

80

16

3

1

10,130

3

b9

3^

lb

0

12.9

13.750

b

bo

bb

9

T
J

"

13.3

i s , 75°

2

*3

b5

9

1

9

"

13*7

16,350

10

bb

U2

b

0

11

»

lb . 2

2b, 100

U

U7

b6

3

0

lb

"

37,950

5

56

33

6

0

18

»

10,650

1

52

35

9

3

Time

Hb

WBC

M

0

12.9

8, boo

2 days

11.85

b

»

6

lb . 8

Case lU
R ab b it l b was n e c r o p s ie d e ig h te e n days a f t e r i n o c u l a t i o n .
isms were c u l t u r e d from th e kidney and u r i n e .

Organ­

The k id n e y s were of averag e

s i z e and e x h i b i t e d s c a t t e r e d w h ite f o c i , e s p e c i a l l y i n th e m edullae.

A

n y e l i t i s was p r e s e n t i n th e l e f t k id n ey .
M icroscopic f i n d i n g s .

The i n t e r s t i t i a l c o n n e c tiv e t i s s u e was

p r e s e n t i n in c r e a s e d amounts i n s c a t t e r e d f o c i i n b o th k id n e y s, e s p e c i a l l y '
i n the c o r t i c e s .

The l e f t kidney showed a s l i g h t amount of desquam ation

of e p ith e liu m on th e t i p o f th e p a p i l l a and a s e v e re p y e l i t i s c h a r a c t e r ­
i z e d by n e c r o s i s of th e p e l v i c mucosa and a zone o f c h ro n ic inflam m atory
changes such a s th e p re s e n c e o f lymphocytes, mononuclear p h a g o cy te s,
a c t i v e f i b r o b l a s t s and a few p s e u d o e o s in o p h ils .

The p e l v i s c o n ta in e d

- 16-

b lo o d , t i s s u e d e b r i s and "bacteriaM

L

p

B

JD

11,200

0

go

lU

6

0

ll.U

13.550

g

70

13

9

0

»

10. g

l b , IhO

lU

U9

29

g

0

6

«

1 1 .1

i h ,950

7

55

29

g

1

9

H

1 2.1

13,*50

6

59

31

3

1

11

"

1 2 .6

l U , 150

3

6U

31

2

0

lU

»

ll.U

lU.Roo

7

Ul

Ul

10

1

lg

"

11.6

1U.350

9

U5

33

12

1

Time

Hb

0

12.5

2 days
U

WBC

Case 15
R a b b it 15 d ie d and was n e c r o p s ie d e i g h t days a f t e r i n o c u l a t i o n .
T h is r a b b i t had a p p e a re d s i c k l y on th e day p r e v io u s to d e a th .
t i n n e d u r i n e h ad s t a i n e d t h e f u r around t h e u r i n a r y o p e n in g .
were c u l t u r e d from th e k id n e y and u r i n e .

Blood
Organisms

The k id n e y s were e n la r g e d and

e x h i b i t e d s e v e r a l s u b c a p s u la r a b s c e s s e s (1 mm d ia m -), - c a p i l l a r y e r o s io n
a n d bloody mucous f l u i d i n th e p e lv e s .
M ic ro sc o p ic f i n d i n g s .

The u r e t e r s were e n la r g e d .

The l i v e r was c o n g e s te d and t h e r e was a

s l i g h t c e n t r a l f a t t y d e g e n e r a tio n .

The lu n g wa3 m ild ly c o n g e s te d and

many p s e u d o e o s in o p h ile s were p r e s e n t i n th e a l v e o l a r w a l l s .
was c o n g e s te d and t h e r e was phn.£-ocytosis of b lo o d p ig m e n t.

Th“ s p le e n
B a c t e r i a were

p r e s e n t i n t i s s u e d e b r i s i n t h e lumen o f t h e u r i n a r y b l a d d e r and th e
e p ith e liu m a p p e a re d s l i g h t l y d e g e n e ra te d .

The u r e t e r a l mucosa was n e c r o t i c

and b a c t e r i a were p r e s e n t i n t i s s u e d e b r i s i n th e lumen.
e x h i b i t e d th e c h a r a c t e r i s t i c p a p i l l a r y e r o s i o n .

The k id n e y s

A b sc e ss e s (1 x 1.5 mm)

c h a r a c t e r i z e d by t i s s u e d e b r i s , p s e u d o e o s in o p h ils and b a c t e r i a were p r e s e n t
i n th e m a d u lla e .

The p e l v e s c o n ta in e d t i s s u e d e b r i s , p s e u d o e o s in o p h ils

-

and 'b a c te r ia -

17-

The p e l v i c mucosa was n e c r o t i c and t h e p e r i p e l v i c t i s s u e

was i n f i l t r a t e d w ith p s e u d o e o s in o p h ils and f i b r i n .
f o c i of lym phocytes, e s p e c i a l l y i n th e c o r t e x .

T h e re were s c a t t e r e d

Some o f t h e p ro x im a l con­

v o l u t e d t u b u l e s e x h i b i t e d clo ud y s w e lli n g .
Time

Hb

WBC

M

L

P

B

E

0

13.9

7.050

0

78

18

u

0

2 days

ll.U

3,800

3

U5

39

13

0

U

«

11.3

6,200

7

Uo

3^

IS

1

6

«

12.9

23,050

10

26

59

u

1

Case l 6
R a b b it 1A was n e c r o p s ie d a s a normal c o n t r o l f o r s e r i e s (A) o f
seven r a b b i t s .

A ll organs were normal e x c e p t th e k id n e y s which e x h i b i t e d

a few sm all w h ite f o c i i n m e d u lla and c o r t e x .
M ic ro sc o p ic f i n d i n g s .

There w ere a few ly m p h o c y tic f o c i s c a t t e r e d

th ro u g h o u t t h e k id n e y s and a few o f t h e c o l l e c t i n g t u b u l e s c o n ta in e d c e l l u ­
l a r d e b ris.
Time

Hb

WBC

M

L

P

B

E

0

11.9

1 1 , 0*50

1

72

26

1

0

Case 17
R ab b it 2A was n e c r o p s ie d one day a f t e r i n o c u l a t i o n .
were c u l t u r e d from th e l i v e r ,

s p le e n , k id n e y and u r i n e .

Ore-a.nisms

The s p le e n

a p p ea re d s w o lle n and d a rk ; th e o t h e r o rg a n s a p p ea re d norm al.
M ic ro sc o p ic f i n d i n g s .

There was i n i n c r e a s e i n p s e u d o e o s in o p h ils

and c o n g e s tio n o f th e lu n g and s p le e n .

The e p ith e liu m on t h e t i p o f t h e

r e n a l p a p i l l a e a p p e a re d s l i g h t l y d e g e n e r a te d .

-1 S -

Time

Hb

WBC

0

1 2 .9

11,300

1 day

13-3

3,750

L

P

B

E

0

73

20

2

0

0

U5

51

U

0

M

Cane 18
R a b b it 3A was n e c r o p s ie d one day a f t e r i n o c u l a t i o n .
( v e r y few) were c u l t u r e d from th e s p le e n and h e a r t b lo o d .

Organisms
A ll o f t h e

organs a p p e a re d normal 9x c e p t f o r a s m a ll w h ite f o c u 9 i n one k id n e y .
M ic ro sc o p ic f i n d i n gs. The lu n g and s p le e n were m ild ly c o n g e s te d
and c o n ta in e d an i n c r e a s e d number of p s e u d o e o s in o p h ils .

One r e n a l m e d u lla

e x h i b i t e d a few sm all f o c i o f p s e u d o e o s in o p h ils .
Time

Hb

0

11.5

1 day

1 1 .8

whC

M

L

P

a

js

lU .650

2

76

IS

3

l

17.350

0

61

36

3

0

Case 19
R a b b it ^A was n e c r o p s ie d two days
were c u l t u r e d from any o f th e o rg a n s.
M ic ro sc o p ic f i n d i n g s .

a f t e r in o c u la tio n .

No o rg an ism s

No l e s i o n s were a p p a r e n t.

There was a s l i g h t i n c r e a s e i n th e number

o f p s e u d o e o s in o p h ils i n th e lung and a s l i g h t c o n g e s tio n o f th e s p le e n .
One s e c t i o n o f th e k id n e y c o n ta in e d a fo c u s (100 x 125 mi e r a )

o f p seu d o ­

e o s i n o p h i l s n e a r t h e s i d e o f th e m e d u lla .
Time

Hb

0

13.1

1 day
2 days

WBC

M

L

P

B

E

12,900

1

75

22

2

0

12.9

13.^50

1

HO

55

6

0

12.1

12,500

2

70

22

5

1

Case 20
R a b b it 5A was n e c r o p s ie d two days a f t e r i n o c u l a t i o n .

Organisms

-lo ­
w ers c u l t u r e d from t h e k id n e y .

AH o rg a n s a p p e a re d norm al ex cep t one

sm all w h ite fo c u s i n c o r t i c o - m e d u l l a r y r e g i o n o f one k id n e y .
M icroscopic f i n d i n g s .

The lu n g was c o n g e s te d , many p s e u d o e o s in o -

p h i l s were p r e s e n t and b a c t e r i a were p r e s e n t i n one of th e l a r g e r b lo o d
v e sse ls.

The s p le e n was c o n g e s te d .

B oth r e n a l m e d u lla s c o n ta in e d s m a ll

a b s c e s s e s (20 0 m lcra d ia m .) c h a r a c t e r i z e d by p s e u d o e o s in o p h ils , t i s s u e
d e b r i s , and l o c a l c o n g e s tio n .
L

p

B

E

2

71

23

2

2

13.050

1

6?

35

2

0

12,100

1

U9

U6

U

0

WBC

Time

Hb

0

1 2 .1

12,300

1 day

ll.U

2 days

1 0 .3

M

Case 21
R a b b it 6a was n e c r o p s ie d t h r e e days a f t e r i n o c u l a t i o n .
were c u l t u r e d from th e s p le e n , k id n e y and u r i n e .

O rganisms

The k id n e y s were o f

normal s i z e b u t e x h i b i t e d w h ite f o c i i n th e c o r t i c e s and m e d u lla s, m e d u lla r
c o n g e s tio n , and a n e c r o t i c fo c u s on t h e t i p of each r e n a l p a p i l l a .
M icro sc o p ic f i n d i n g s .
p s e u d o e o s in o p h ils .

The lu n g c o n ta i n e d an i n c r e a s e d number o f

The s p le e n was c o n g e s te d and showed in c r e a s e d phago­

c y t o s i s o f b lo o d p ig m e n t.

There were f o c i i n t h e u r i n a r y b l a d d e r mucosa

which l i k e l y r e p r e s e n t e d e a r l y l e s i o n s i n t h e b l a d d e r .

These were

c h a r a c t e r i z e d by some desquam ation and c a v i t y f o r m a tio n of t h e e p i t h e l i u m ,
h y a l i n e b o d ie s i n some e p i t h e l i a l c e l l s , c o n g e s tio n of t h e mucosa and
i n f i l t r a t i o n w ith s e r o u s f l u i d , f i b r i n and p s e u d o e o s in o p h ils .

I n th e

s e c t i o n of th e r i g h t k id n e y t h e r e was a fo c u s a t t h e t i p of th e p a p i l l a
c h a r a c t e r i z e d by n e c r o s i s , masses o f b a c t e r i a , c o n g e s tio n , and i n f i l t r a t i o n
by p s e u d o e o s in o p h ils .

A lso p r e s e n t i n th e m edulla were s e v e r a l sm a ll f o c i

c h a r a c t e r i z e d by p s e u d o e o s in o p h ils i n th e t u b u le s and c o n g e s tio n .

-2 0 -

Hb

Time

WBC

M

L

P

B

E

0

12.9

13.350

1

60

31

g

0

1 day

12.9

15.850

2

22

69

7

0

2 days

1 1 .1

11,100

7

52

35

6

0

3

1 1 .U

1 6 ,1 50

0

52

U2

5

1

"

Ca.3e 22
R ab b it 7A was n e c r o p s i e d t h r e e days a f t e r i n o c u l a t i o n .
were c u l t u r e d from t h e k id n e y and u r i n e .

Organisms

The k id n e y s w ere o f normal s i z e

and e x h i b i t e d m e d u lla r c o n g e s tio n w ith a n e c r o t i c fo c u s on t h e t i p of
each r e n a l p a p i l l a .
M ic ro sc o p ic f i n d i n g s .

The lu n g e x h i b i t e d m ild c o n g e s tio n and

edema w ith an i n c r e a s e i n p s e u d o e o s in o p h ils .

The s p le e n was c o n g e s te d .

The l e s i o n s a t th e t i p s of t h e r e n a l p a p i l l a e were v e r y s i m i l e r t o th o s e
i n r a b b i t 6a .

The r e n a l m e d u lla s were m arkedly c o n g e s te d and c o n t a i n e d

s e v e r a l f o c i o f p s e u d o e o s in o p h ils .

The p e l v i c e p ith e liu m showed s l i g h t

d e g e n e r a tiv e changes where i t c o n ta c te d th e p a n i l l a r l e s i o n .
Time

Hb

0

12.1

13,600

1 day

11.6

2 days
3

"

WBC

M

L

P

B

E

2

78

15

5

0

n .U o o

0

56

Ul

3

0

11.2

13,600

0

52

Uo

g

0

11.0

11,1*50

2

69

26

3

0

Case 23
R a b b it Sa was n e c r o p s i e d f o u r days a f t e r i n o c u l a t i o n .
were c u l t u r e d from th e l i v e r , h e a r t b lo o d , kidne;/- and u r i n e .
a p p e a re d s l i g h t l y e n la r g e d .

Organisms
The s p le e n

I n th e h e a r t was a sm all r e d nodule on b a s e

o f th e a o r t i c v a l v e and a s m a ll (1 mm x 2 mm) thrombus a t t a c h e d to t h e
base of th e l e f t a t r io v e n tr ic u la r v a lv e .

The k id n e y s w ere o f a v e r a g e s i z e

-2 1 -

and e x h i b i t e d a n e c r o t i c a r e a on th e t i p o f each r e n a l p a p i l l a *

The

u r e t e r s a p p e a re d v e r y s l i g h t l y c o n g e s te d .
M ic ro sc o p ic f i n d i n g s .
edema.

The lu n g showed a s l i g h t c o n g e s tio n and

The b l a d d e r e p ith e liu m appeared s l i g h t l y d e g e n e r a te d .

The

u r e t e r a l e p ith e liu m a p p e a re d d e g e n e ra te d and th e lumen c o n ta in e d b lo o d
and b a c t e r i a .

The l e s i o n s i n th e r e n a l p a p i l l a e and m e d a lla e were s im i­

l a r to th o s e i n r a b b i t 6A on ly much more e x t e n s i v e .

The t»elvic e p i ­

th e liu m showed d e g e n e r a tiv e changes where i t con tacted , th e p a p i l l a r
lesio n .

T here was a s l i g h t i n f i l t r a t i o n o f f i b r i n i n t o th e p e r i p e l v i c

t is s u e in th e se p la ce s.
Time

Hb

0

12.3

1 day

WBC

M

L

p

B

E

10,950

0

71

25

H

0

12.3

11,000

1

U5

51

3

0

2 days

10.5

13,850

0

36

56

7

1

3

"

io . U

12,600

0

73

23

H

0

U

"

11.8

20,750

-

22-

Table I
Summary of Kidney L e s i o n s and C u l t u r e R e s u l t s

Time

Kidney
L e s io n s

Culture (1)

1

1 /2 h r .

-

+ h b l , l v , 1 , kd

2

1 clay

-

+ I v , kd

3

2 days

+

-

H

3

"

+

+ kd, u r

5

U

"

+

+ h b l , kd, u r

6 (?)

5

"

+

+ h b l , kd, s p l , bm,

7

6

"

+

+ kd, u r

2 (3)

7

"

+

+ kd

9

9

»

+

+ kd, u r

10

11

»

+

+ kd, u r

li

17

"

+

+ kd, u r

12

17

H

+

-

13

18

»

+

+ kd, u r

lU

18

"

+

+ kd, u r

13 (? )

8

»

+

+ kd, u r

1A

0

-

2A

1 day

-

+ s p l , l v , kd, u r

3A

1

-

+ hbl, sol

Ua

2 days

-

-

5A

2

11

+

+ kd

6a

3

"

+

+ s p l , kd, u r

7A

3

"

+

+ kd, u r

3A

U

»

+

+ h b l , l v , kd, u r

R ab bi t

"

( 1 ) c u l t u r e s ; 1- l u n g , l v - l i v e r , bm-■bone marrow, s p l - s p l e e n ,
k d - k id n e y , u r - u r i n a r y b l a d d e r , h b l - h e s r t b l o o d .
( 2 ) Died
( 3 ) Moribund

-

23“

Tftble I I
Averages of Hemn t o l o g i c R e s u l t s
No. An ( ? )

Hb (3)

17BC (U)

M.<51

0

15

12.6

7.893

0.8

2

13

11.0

7.927

U.5

u

11

11.3

15, 5^5

6

9

12.2

9

6

11

P

B

1

ro
•

rB (1)

23.3

3-5

o.U

56.2

31*7

7.5

0.1

9 .2

Hq.O

32.8

8.5

0.5

lU,S7?

7-3

H5.

Uo.

6.H

0.8

12.7

21,366

6.3

U8.7

Hi.

3-8

0.2

5

12.7

I P , 610

6.2

U7.6

Hi. 2

5.

0

lU

U

13.0

19,662

3-3

^5. U 77.

9-

0.3

17

2

1U.3

13.575

9 .0

77.

Hq.

U.

1.0

IS

2

13.2

17,100

5-0

Uq.

3U.

10.

2.0

0

S

12.U

12,512

1.1

72.6

22.5

3-U

O.U

1

7

12.3

13.050

0-7

^7-3

Up .

U.

0

2

5

11.0

12,630

2.0

51.8

39.8

6.

O.U

3

3

10.8

13,016

0.7

65.

30.3

u.

0.3

U

1

11.8

20,750

-

-

L

-

-

-

( l)

Lays a f t e r i n o c u l a t i o n .

(2)

Number of a n im a ls i n t h e group u n d e r c o n s i d e r a t i on a t t h a t t:

(3)

Hemoglobin I n grams p e r 100 ml o f "blood.

(U)

Le u c o c y te s p e r cu"bic mm.

(5)

P e r c e n t o f monocytes, lymp hoc ytes, p s e u d o e o s l n o o h i l s , "basophils
and e o s i n o p h i l s .

-?U -

DISCTJSSION
The l e s i o n s o f e x p e r i m e n t a l p y e l o n e p h r i t i s i n t h e r a h h i t a r e
v e r y s i m i l a r t o t h o s e fo und i n spontan eo us c a s e s of p y e l o n e p h r i t i s i n
c a t t l e a s d e s c r i b e d o r i g i n a l l y by F r n s t (1905) and l a t o r by o t h e r s ( J o n e s
and L i t t l e , 1925; Thorp e t a l , 19U5).

The l e s i o n s a r e a l s o s i m i l a r t o

t h o s e found i n e x p e r i m e n t a l p y e l o n e p h r i t i s i n mice by L o v e l l and C o tc h in
(19H6).
The f a c t t h a t t h e c o n d i t i o n i n t h e r a b b i t wa.s p r o d u c e d by i n t r a ­
venous i n j e c t i o n of £. r e n a l e shou ld n o t be c o n s i d e r e d a s e v id e n c e t h a t
t h e s pon tan eo us d i s e a s e i n c a t t l e i s of hematogenic o r i g i n .

The weight

of e vid enc e i n t h e ca se of t h e cow would i n d i c a t e t h a t i t i s an a s c e n d i n g
ty p e of i n f e c t i o n which may g a i n e n t r a n c e by means o f v u l v a r or u r e t h r a l
c o n t a m i n a t i o n ( J o n e s and L i t t l e ,

l l ? 6 ; F e e n s t r a and Thorp, 19hg).

F i s e n d r a t h and S c h u l t z (1917) have d e m o n s t r a t e d , e x p e r i m e n t a l l y i n r a b b i t s
and dogs, t h e p r e s e n c e o f an an astomosing network of lynvnhatics i n t h e
w a l l of th e b l a d d e r and of t h e u r e t e r which communicates above w i t h a
s i m i l a r l y m p h a t i c network i n t h e r e n a l r e l v i s and -parenchyma and below
w i t h th e lympha+ics of t h e p e l v i c s t r u c t u r e s i n b o t h t h e male and f e m a le .
They s u g g e s t t h a t t h i s i s t h e r o u t e of an a s c e n d i n g i n f e c t i o n o f t h e
urinary tra c t.
T hi s r e p o r t s h ou ld n o t be t a k e n as im ply in g t h a t o nl y C. r e n a l e
c a u s e s l e s i o n s of p y e l o n e p h r i t i s i n t h e r a b b i t .

Dudgeon and Goadby (1950)

r e p o r t e d t h a t a s u p p u r a t i v e p y e l o n e p h r i t i s f o l l o w e d t h e i n t r a v e n o u s in o c u ­
l a t i o n of St a p h y lo c o c cu s a u r e u s i n t o r a b b i t s .
The f a t e of th e organisms from t h e time of i n j e c t i o n u n t i l t h e
b e g i n n i n g of l e s i o n f o r m a t i o n i n t h e k id n e y i s o f i n t e r e s t .

The d a t a i n

T a b le I would i n d i c a t e t h a t , i n g e n e r a l , t h e organisms were found i n i t i a l l y
i n t h e lu n g , s p l e e n and l i v e r and l a t e r i n th e k id n e y and u r i n e .

The

-

25-

i n t e r v a l "between n e c r o p s i e s wan too g r e a t t o o b t a i n a more a c c u r a t e p i c ­
t u r e of t h e r o u t e o f t h e org anisms t h r o u g h t h e animal body.

A study of

t h e r e s u l t s o f i n t r a v e n o u s i n o c u l a t i o n of o t h e r organisms by v a r i o u s
worke rs may c a s t more l i g h t on t h e p r o c e s s .

B u l l (191*0 showed t h a t

v i r u l e n t s t r e p t o c o c c i and pneumococci t e n d e d t o d i s a p p e a r from t h e c i r c u ­
l a t i n g b lo o d of r a b b i t s i n two t o f o u r h o u r s and t o r e a p n e a r a t f i v e t o
s i x hours and e v e n t u a l l y c au s e d e a t h .

L e s s v i r u l e n t orga nis ms may not

r e a p p e a r o r may cauee a c h r o n i c i n f e c t i o n .

B a r t l e t and Ozaki (1917)

r e p o r t e d t h a t " j ' i c r o c o c cus a u r e u s 11 i n j e c t e d i n t o t h e l e f t v e n t r i c l e of
dogs was a lm o s t im me dia te ly s t o r e d i n t h e lu n g c a p i l l a r i e s , b e i n g a t
f i r s t c h i e f l y e x t r a c e l l u l a r and th e n r a p i d l y i n g e s t e d by p o ly m or ohonu cle a r
leukocytes.

S h o r t l y a f t e r i n j e c t i o n t h e c o c c i i n c r e a s e d i n number i n t h e

l i v e r and s p l e e n a s they d e c r e a s e d i n number i n t h e lu n g .

The b a c t e r i a

c o m p l e t e l y d l s a o p e a r e d from t h e s e o r g a n s w i t h i n bg t o 72 h o u r s .

The

c occi d id no t p r i m a r i l y lo d g e i n th e k i d n e y i n any c o n s i d e r a b l e numbers,
b u t were s e c o n d a r i l y d e p o s i t e d t h e r e a f t e r a r e r i o d 0f a d a p t a t i o n e l s e ­
where ( c o c c i were found i n t h e kid ne y a t i f h ou rs b u t not a t e i g h t h o u r s ) .
T h e i r l o c a t i o n h e r e was p r o b a b l y no t em bo lic .

Hopkins and P a r k e r ( 191?)

showed t h a t i n j e c t e d h e m o l y t i c s t r e p t o c o c c i te n d ed t o d i s a p p e a r from t h e
b lo o d i n about two h o ur s and t o r e a p p e a r i n i n c r e a s i n g numbers a f t e r f o u r
and o n e - h a l f h o u r s .

I n c a t s and r a b b i t s t h e orga nism s withdrawn from t h e

c i r c u l a t i o n were found main ly i n t h e l u n g , s p l e e n and k i d n e y .

Nagao ( 1920)

i n j e c t e d k i l l e d n o n -h e m o ly ti c s t r e p t o c o c c i i n t o g u in e a p i g s and o b s e r v e d
t h a t t h e org anisms a cc um ula te d alm os t i m m e d ia te ly i n t h e l u n g c a p i l l a r i e s
i n s i d e po ly mo rp ho nucle ar l e u k o c y t e s and l a t e r i n t h e r e t i c u l o - e n d o t h e l i a l
c e l l s of t h e l i v e r and s p l e e n .

Dudgeon and Q-oadby ( 1971) r e n o r t e d t h a t

i n t r a v e n o u s l y i n j e c t e d S ta p h y lo c o c c u s a u r e u s organisms were h e l d i n t h e
l u n g s where th e y were a c t i v e l y p h e g o c y to s e d by t h e p o ly m o r p h o n u c le a r

- 26-

leukocytes w ith in f iv e minutes.

S u b s e q u e n t l y , t h e c o c c i were d i s t r i b u t e d

t o t h e o t h e r org a ns such a s t h e l i v e r and s p l e e n and t o t h e k id n e y .

It

may be c o n c l u d e d from s t u d y of t h e above mentioned r e s u l t s t h a t , i n
g e n e r a l , t h e b a c t e r i a t r a v e l f i r s t t o t h e lu n g and l a t e r a r e r e l e a s e d t o
be p i c k e d up by th e l i v e r and s p l e e n and l a t e r may b e found i n t h e k i d n e y .
The mechanism o f i n f e c t i o n i n t h e kid n e y i s of s p e c i a l i n t e r e s t .
L o v e l l and C otc h in (19*^6) have made o b s e r v a t i o n s on t h i s p r o c e s s i n t h e
mouse k id n e y .

They f i r s t saw th e organisms i n clumps i n the c a o i l l e . r i e s

between t h e c o l l e c t i n g t u b u l e s n e a r t h e apex of t h e p a p i l l a .

T hi s was

accompanied by l o c a l d i l a t i o n o f c a p i l l a r i e s w i t h an a c c u m u l a t i o n of
po ly m o r p h o n u cl e a r l e u k o c y t e s which l a t e r p a s s e d thr ough t h e normal or
n e c r o t i c e p i t h e l i u m of t h e c o l l e c t i n g t u b u l e s i n t o t h e i r lumens.

Small

a b s c e s s e s d e v el ope d i n t h e p a p i l l a o f t e n j u s t under t h e v i s c e r a l p e l v i c
epithelium.

Pus was found i n t h e r e n a l p e l v i s as e a r l y a s t h i r t y h o u r s .

N e c r o s i s o f t h e p a p i l l a exten de d towa rd t h e c o r t e x .

L a rg e cluwos o f gram

p o s i t i v e b a c t e r i a were seen i n t h e n e c r o t i c m a t e r i a l w i t h a zone of
p s e u d o e o s i n o p h i l s a t t h e e x t e n d i n g edge of th e n e c r o t i c a r e a .

Pseudo­

e o s i n o p h i l s were a l s o found, i n s t r e a k s up t h e medulla towa rd and i n t o t h e
cortex.

The r e n a l p e l v i s was d i l a t e d w i t h p u r u l e n t m a t e r i a l .

The p e l v i c

e p i t h e l i u m showed a c u t e in fl am m a to ry chang es w i t h a b s c e s s and u l c e r forma­
tion.

The p a t h o g e n e s i s o f e x p e r i m e n t a l p y e l o n e p h r i t i s i n t h e r a b b i t , a s

r e p o r t e d i n t h i s p a p e r was e s s e n t i a l l y s i m i l a r to t h a t d e s c r i b e d i n t h e
c a s e of t h e mouse.

The e a r l i e s t change i n t h e ki dn e y was t h e accumula­

t i o n of p s e u d o e o s i n o p h i l s i n t h e c a p i l l a r i e s and c o l l e c t i n g t u b u l e s w i t h
congestion in the p a p i l l a r y region.

B a c t e r i a may have b e e n p r e s e n t i n

t h e s e p s e u d o e o s i n o p h i l s b u t t h e y co uld no t b e c l e a r l y i d e n t i f i e d .

Fo ll ow ­

i n g t h i s , a t t h r e e and f o u r days a f t e r i n o c u l a t i o n , n e c r o s i s and e r o s i o n
o f t h e p a p i l l a were found.

Masses of b a c t e r i a were see n i n th e r e m a in s

-? 7 o f c a p i l l a r i e s and t u b u l e s , and in t h e d e b r i s i n t h e p e l v i s a s has be en
n o t e d i n t h e s tu d y of b o v in e p y e l o n e p h r i t i s ( P e e n s t r a a nd Thorp, 19U6).
Psexido e o s i n o p h i l s were numerous in a n d a ro u n d t h e n e c r o t i c l e s i o n .
p s e u d o e o s i n o p h i l s a pp ea re d, i n many c a s e s ,

The

to c o n t a i n b a c t e r i a b u t t h e

g r a n u l a r i t y of the c e l l s and f a i n t s t a i n i n g q u a l i t y of any b a c t e r i a
i n s i d e them i n t e r f e r e d w it h p r e c i s e o b s e r v a t i o n s .

S t r e a k s o f pseudo­

e o s i n o p h i l s were found i n t h e me du lla e x t e n d i n g toward t h e c o r t e x .

At

t h i s s t a g e ( a b o u t f o u r days) the p e l v i c mucosa a p pe ared n e c r o t i c and t h e
p e r i p e l v i c t i s s u e s c o n t a i n e d serous f l u i d ,
pseudoeosinophils.

fibrin,

e r y t h r o c y t e s and

Lymphocytes s m e a r e d on a b c u t t h e s i x t h day and

mononuclear p h a g o c y te s and a c t i v e f i b r o b l a s t s a pp ear ed on th e s e v e n th
day.

These l a t t e r t h r e e c e l l s i n c r e a s e d i n number a s ti m e p a s s e d w h il e

t h e p s e u d o e o s i n o p h i l s de c r e a se d in number but nev er d i s a p p e a r e d e n t i r e l y .
L a t e r (17 a n d 18 d a y s ) , changes in t h e c o r t e x such a s an i n c r e a s e i n
i n t e r s t i t i a l c o n n e c t i v e t i s s u e , d i l a t i o n and c o n s t r i c t i o n of t u b u l e s and
a c c u m u l a t i o n of lymphocytes were f o u n d .

I n t h e s e more c h r o n i c c a s e s t h e

p a p i l l a r y e r o s i o n seemed t o be p r o g r e s s i n g v e r y slow ly i f a t a l l b u t t h e
p y e l i t i s appeared to be increasing i n s e v e r ity .

N e c r o s i s and in fl a m m a to ry

changes r e s e m b l i n g t h e p y e l i t i s to a g r e a t e x t e n t were fo und i n t h e u r e t e r s
s h o r t l y a f t e r t h e a p p ea ra n ce of th e p y e l i t i s .
As may be e x p ec te d i n animal e x p e r i m e n t a t i o n , t h e r e were i n d i ­
v i d u a l v a r i a t i o n s i n r e a c t i o n to i n o c u l a t i o n and t h e r e a c t i o n i n s e r i e s A
was s l i g h t l y d i f f e r e n t from the o t h e r group .

For t h i s r e a s o n mice c o u l d

be used t o a dva n ta g e b e c a u s e th e re i s l e s s c a r e and expense c o n n e c te d
w i t h t h e u s e o f g r e a t e r numbers.
As may be s een i n Table I I t h e r e was a s l i g h t i n i t i a l drop i n
t h e hemoglobin v a l u e s f oll ow e d by a n i n c r e a s e .

This c o u l d b e due to t h e

f a c t t h a t d u r i n g the f i r s t week a f t e r i n o c u l a t i o n t h e f u n c t i o n o f t h e

-2 8 bone marrow was concer ne d mainly w it h p r o d u c t i o n of g r a n u l a r l e u k o c y t e s
and t h e re b y t h e p r o d u c t i o n of e r y t h r o c y t e s was hampered.
I n b o t h s e r i e s of r a b b i t s i t was on t h e f o u r t h day t h a t t h e b l o o d
l e u k o c y t e co unt showed a marked i n c r e a s e .

The s l i g h t i n i t i a l drop i n t h e

numbers of l e u k o c y t e s i s l i k e l y due t o t h e f a c t t h a t many of them a r e
t e m p o r a r i l y lo d g e d i n t h e pulmonary c a p i l l a r i e s and i n o t h e r o r g a n s .
Ae might b e e x p e c te d , t h e r e was an i n c r e a s e i n t h e p e r c e n t a g e of
p s e u d o e o s i n o p h i l s i n t h e blood and a d e c r e a s e i n th e p e r c e n t a g e o f lympho­
cytes.

I n t h e c a s e of t h e p s e u d o e o s i n o p h i l s , immature forms were o f t e n

seen a s t h e p e r c e n t a g e i n c r e a s e d b u t were n o t r e c o r d e d a s such be ca us e
of th e d i f f i c u l t y i n p r e c i s e i d e n t i f i c a t i o n of t h e s e forms due to t h e
b r i l l i a n t l y r e d s t a i n i n g aranuJ.es p r e s e n t .
p e r c e n t a g e of monocytes.

There was an i n c r e a s e i n t h e

Less e x p e c te d and n o t e a s i l y e x p l a i n a b l e was

t h e i n c r e a s e i n p e r c e n t a g e of b a s o p h i l s .

The main d i f f i c u l t y i s t h a t

l i t t l e or n o t h i n g i s known of the f u n c t i o n of t h e b a s o p h i l .

There was

no t a s i g n i f i c a n t or me an in gfu l change i n t h e p e r c e n t a g e o f e o s i n o p h i l s .
Summary
The p a t h o g e n e s i s of e x p e r i m e n t a l p y e l o n e p h r i t i s i n r a b b i t s p r o ­
duced by i n t r a v e n o u s i n o c u l a t i o n o f C ory nebacterlum r e n a l e has been
d e s c r i b e d and d i s c u s s e d .
and s e v e r a l d i e d .

Host i n o c u l a t e d r a b b i t s showed ki d n e y l e s i o n s

The l e s i o n s i n th e k i d n e y c o n s i s t e d of a p a p i l l i t i s

and p y e l i t i s c h a r a c t e r i z e d by n e c r o s i s and a c u t e i n fl a m m a to ry changes
e a r l y i n t h e c o u r s e and by c h r o n i c in f l a m m a to r y changes l a t e r i n t h e
course.

B a c t e r i a were p r e s e n t mainly i n t h e n e c r o t i c d e b r i s of t h e

l e s i o n s and i n t h e p e l v i s .
The l e s i o n s i n t h e r a b b i t ki dn e y were s i m i l a r t o t h o s e found i n
p y e l o n e p h r i t i s i n the cow.

The fact, t h a t t h e e x p e r i m e n t a l p y e l o n e p h r i t i s

-

29-

was of he m at ogeni c o r i g i n i n no way n e g a t e s t h e g e n e r a l l y a c c e p t e d t h e o r y
o f an a s c e n d i n g i n f e c t i o n i n the cow.
The r a b b i t may be u s e d , a d v a n t a g e o u s l y , i n t h e s t u d y of p y e l o n e p h r i t i s and a l s o i n t h e s t u d y of t h e p a t h o g e n i c and a n t i g e n i c p r o p e r t i e s
of C ory nebacterium r e n a l e .

-

30-

REFEP.ENCES

B a r t l e t t , C. J - a n d C z a k i , Y. 1917 The f a t e o f M i c r o c o c c u s a u r e u s
i n t r o d u c e d i n t o t h e b l o o d s t r e a m o f d o g s . J . Med. R es . 35»
H65-*!S'6 .
B e r g e y , D. H . , e t a l .
1939 Manual o f d e t e r m i n a t i v e B a c t e r i o l o g y .
5 th ed.
The W i l l i a m s a n d W i l k i n s C o . , B a l t i m o r e .
B u l l , C. G.
191*+ A method f o r e s t i m a t i n g t h e " b a c t e r i a i n t h e c i r c u ­
l a t i n g b l o o d i n r a b b i t s . J . Exp. Med., 20, 237-2*48.
R o l l , E. R.
19*4-2 TTnoublished d a t a .
S ta te College.

Rent,

o f Animal P a t h o l o g y ,

Michigan

Dudgeon, L. S. a n d Goadby, H. K. 1930 F u r t h e r o b s e r v - ' t i on s on t h e
s t a p h y l o c o c c i w ith s p e c i a l r e f e r e n c e to t h e i r h e m o ly s is and
v ariab ility .
J . Hyg. , 3("h ISO-199.
Dudgeon. L. S. and Goadby, H. K. 1171 The e x a m i n a t i o n o f t h e t i s s u e s
and some o b s e r v a t i o n s on t h e blood, p l a t e l e t s o f r a b b i t s a t
i n t e r v a l s of f i v e m inutes and l a t e r a f t e r in t r a v e n o u s i n o c u l a t i o n
of S t a n h y 1 0c o c c u s a u r e u s and I n d i a i n k .
J . H y g ., 31* 2^7-296.
E i s e n d r a t h , R. N. and S c h u l t z , 0. T.
1917 The p a t h o f i n v o l v e m e n t i n
ascending i n f e c t i o n of the u rin a ry t r a c t .
J . Med.. R e s . , 3C' ,
209- 1 3 9 .
E n d e r l e n , R.
1891 Z. T i e rm e d . , 17, 7^6.
C i t e d by R i t z e n t h a l e r , M,
1910 The p a t h o l o g y o f b a c i l l n r y p y e l o n e p h r i t i s i n c a t t l e .
J . Comp. P a t h . T h e r a n . , 23, 33~5Q*
E r n s t , W. 1909 Z e n t r . B a k t . P a r a s i t e n k . ,
by R i t z e n t h a . l e r ( s e e E n d e r l e n ) .

71, 5*49-666; Up, j q .

cited

F e e n s t r a , E. S. , C l a r k , C. F . and T h o r p , F r a n k , J r .
11*49 A r t i f i c i a l l y
i n d u c e d c a s e o f p y e l o n e p h r i t i s i n a b o v i n e . M.S.C. V a t . , 9 , 1*17- 1 9 0 .
F e e n s t r a , E. S. a n d Thor n, F r a n k , J r .
11*46 B a c t e r i o - p a t h o l o g y of
i n f e c t i o u s bovine p y e l o n e p h r i t i s .
Am. J . V e t. R e s . , 7. *432-^76.
Hoffman, W. S. 19*41
and. Co. , N.Y.

P h o telo m etric c l i n i c a l chem istry.

W i l l i a m Morrow

H o p k in s , J . G. a n d P a r k e r , J . T. 1118 The e f f e c t o f i n j e c t i o n s o f
h e m o l y t i c s t r e p t o c o c c i on s u s c e p t i b l e and i n s u s c e p t i b l e a n i m a l s .
J . Exp. Med., 27, 1 - 2 6 .
J o n e s , F. S. a n d L i t t l e , R. B. 1975 S p e c i f i c i n f e c t i o u s c y s t i t i s
p y e l o n e p h r i t i s o f cows. J . Exo. Med., U?, 597-6o7*
J o n e s , F. S. and L i t t l e , R. B. 1926 The o r g a n i s m a s s o c i a t e d ^ i t h
s p e c i f i c i n f e c t i o u s c y s t i t i s a.nd p y e l o n e p h r i t i s o f cows. J . Exp.
Med., UU, 11*.20.

and

-3 1 L o v e l l , R.
19H6 S t u d i e s on Corynei>acteriurn r e n a l e . I . A s y s t e m a t i c
s t u d y o f a number o f s t r a i n s .
J . Como. P a t h . , 9 6 , 196-20U.
L o v e l l , R. a n d C o t c h i n , P.
Ihhfi S t u d i e s on Co r y n e b a c t e r i u rn r e n a l e .
II.
The e x p e r i m e n t a l p a t h o g e n i c i t y f o r mice. J . Comp. P a t h . ,
5 6 , 205- 21U.
Nagoa., K. 1920 The f a t e o f k i l l e d n o n - h e m o l y t i c s t r e p t o c o c c i i n j e c t e d
i n t o t h e "blood a nd t h e r e s u l t i n g c e l l u l a r c h a n g e s .
J. Infect.
L i s . , 27, 1 2 7-3 6 2.
T h o r n , P r a n k , J r . , T.angham, R. P . , C l a r k , C. P. and L o l l , P. R.
I'bh}
The p a t h o l o g y of "bovine p y e l o n e p h r i t i s .
Am. J . V e t . R e s . , H,
2UO-2U9.

F ig .

F ig .

1.

2.

R abbit p y e lo n e p h r iti3 a t n e cro p sy .
T his an im al
d ie d a f t e r a s e r i e s of i n o c u la tio n s w ith c u l t u r e
ITo. 2 ( T h e s i s , P a r t I I ) .
The d i g e s t i v e s y s t e m
h a s "been r e m o v e d .
Th e k i ^ n e T r g a r e e n l a r g e d a n d
e x h ib it w hite f o c i .
The u r e t e r s a r e much e n l a r g e d
The u r i n a r y b l a d d e r i s d i s t e n d e d w i t h b lo o d y u r i n e
C o l o r p r i n t fro m Kodaehrom e n e g a t i v e .

R a b b it k id n e y , show ing p a p i l l a r y n e c r o s i s an d
erosion.
This anim al d ie d a f t e r a s e r i e s of
i n o c u l a t i o n s w i t h c u l t u r e Ho. 5 ( T h e s i s , P a r t I I ) .
C o lo r p r i n t fro m Kodaehrome n e g a t i v e .

3*

SaTie

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size.

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kidneys,

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m e d u l l a end e a r l y mart i 11a r y n e c r o s i s .

inocul
in

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F ig.

6.

R a b b it k id n e y s , removed a t d e a t h s i x days a f t e r
i n o c u l a t i o n , ghow ing e n la r g e m e n t, p a p i l l a r y
n e c r o s is and w h ite s tr e a k s in the m edulla and
co rtex .

F ig.

n B k

7*

R a b b it k i d n e y s , rem oved a t d e a t h se v e n day s
a f t e r in o c u l a t i o n , shoving e n la rg e m e n t,
p a p i l l a r y n e c r o s is and w hite s tr e a k s in th e
m e d u lla and c o r t e x .

P ig.

S.

C ro s s s e c t i o n o f k id n e y from r a 'b M t
w hich
was n e c r o p s i e d on t h e s e c o n d day a ^ t e r i n o c u ­
la tio n .
No l e s i o n s a r e e v i d e n t .
The p e l v i c
m u c o s a w as l o s t f r o m t h e s p e c i m e n .
Gram—
W eigert s t a i n .
X9.

.

V
i.

I u jS S - w

J

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a

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r

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P ig.

9*

C r o s s s e c t i o n o f k i d n e y f r o m r a b b i t 6a w h i c h
w as n e c r o p s i e d t h r e e d a y s a f t e r i n o c u l a t i o n .
A s l i g h t amount o f - n a p i l l s r y n e c r o s i s i s
evident.
F i ^ s . l 6 a n d 23 sh ow some o f t h e s e
c h a n g e s i n more d e t a i l .
H em ato x y lin and
eo sin sta in .
X9 -

F v v ? :.’ .'- ? : v ; - ; : f

F i g.

10.

O r o s s s e c t i o n o f k i d n e y ^rom r a ^ h i t
w h i c h wan
necropsied. fo u r days a f t e r i n o c u l a t i o n .
P ap illary
n e c r o s i s a n d e r o s i o n , a n d t h e h « f r i n n i n y o*“ s t r e a k s
(■ D seudoeosinoohils) in th e m e d u lla a r e e v i d e n t .
F i y . 17 s h o w s som e
t h e s e c h a n y e s i n more d e t a i l
O-ram- We isre rt s t a i n .
X9.

F ig.

11.

S a g i t t a l s e c t i o n o f k i d n e y f r o m r a b b i t *A.
P a p i l l a r y n e c r o s is and e ro s io n , and a s tr e a k
o f p s e u d o e o s in o p h ils e x te n d in g from th e
o a p i l l a to the c o r te x a r e e v id e n t.
Fig:. 18
s h o w s some o f t h e s e c h a n g e s i n m o r e d e t a i l .
G -r a m - W e i g e r t s t a i n .
X9.

? r tS*’• J*
V 's
?

1-

f,

F ig.

12.

C r o s s s e c t i o n o f a k i d n e y fro m a ra'b 'b lt w i t h a
c h r o n i c form o f -n y e lo n e p h riti a.
T his anim al had
"been g i v e n a s e r i e s o f t e n w e e k l y i n o c u l a t i o n s
w i t h c u l t u r e >To. 5 ( T h e s i s , P a r t I I ) .
P y elitis
and - p a p i l l i t i s a r e e v id e n t.
F i a s . 15, 20 and
s ho w some o f t h e s e c h a n g e s i n g r e a t e r d e t a i l .
H em atoxylin and 9 o s in s t a i n .
X9.

P ig.

13.

P y e l i t i s and D a p l l l i t i a in th e k idney of r a b b i t 9
■which w a s n e c r o p s i e d n i n e d a y s a f t e r i n o c u l a t i o n .
The p a p i l l i t i s seems t o b e re a re s B in iT w h i l e t h e
m y e litis i s s t i l l very severe.
Gram-Weip-ert s t a i n .
X^5.

F ig.

lU.

P y e l i t i s a n d p a p i l l i t i s i n t h e k i d n e y o f r a b b i t 10
w h ic h was n e c r o p s i e d e l e v e n d a y s a f t e r i n o c u l a t i o n .
T h e d a r k m a s s e s i n t h e p e l v i s a r e made up o f
b acteria.
G ram -W eigert s t a i n .
XU5 .

SSfB

F ig.

15.

C h ro n ic form
p y e l i t i s and n a p i l l i t i a .
F i g . 12
i n c l u d e s t h i s f i e l d a n d F i g s . 2 0 a n d 25 show som e
o f t h e s e c h a n g e s i n more d e t a i l .
H em atoxylin and
eosin s ta in .
X55.

s

4

; *'

•.• -I:

\

P ig.

16.

P a p illitis.
T h e d a r k m a s s e s a r e made up o f
"bacteria.
P ig . 9 co n tain s t h i s f i e l d .
P i e . 23
s h o w s t h e " b a c t e r i a a n d t u b u l e s i n mo re d e t a i l .
G-ram-Weigert s t a i n .
X l60.

P ig.

17.

P a p illitis.
B a c t e r i a may "bo s ° e n i n t h e l o w e r
h a l f o f t h i s p h o t o m i c r o g r a p h w h i l e many p s e u d o ­
e o s i n o p h i l s a r e p r e s e n t " b et w e e n t h e t u b u l e s a n d
in the n e c ro tic d e b ris.
P i g . 10 c o n t a i n s t h i s
field .
G ram -W eigert s t a i n .
X l6 o .

F ig.

IS*

P ap illitis.
C o llectin g tu b u les, n e c ro tic tis s u e
a n d b a c t e r i a may b e s e e n .
F i g . 11 c o n t a i n s t h i s
field .
G-ram-Weigert s t a i n .
X l60.

r- t • > f * . *■
> ,.

.

s.* ^
j:

r

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19.

P a p i l l i t i s i n k i d n e y o f r a b b i t 6 w h ic h was
n e c ro p sie d f i v e days a f t e r in o c u la tio n .
M asses
o f b a c t e r i a may b e s e e n i n t h e c a p i l l a r i e s a n d
c o lle c tin g tu b u les.
Gram-VTeigert s t a i n .
X l60.

-S ..^S ?- -rti^r-ej*-!^* - -O vV

■ *<«,

jf
f t

3
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r . 9 1 .

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ik'^ L

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'V*.

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'?* r „ IQ

P a p i l l i t i s , showing' n e c r o s i s , a c c u m u l a t i o n o f
inflam m ato ry c e l l s and fo rm a tio n of c r y s t a l s .
P i g s . 12 a n d 15 c o n t a i n t h i s f i e l d .
P i g . 25
shows t h e c r y s t a l s i n g r e a t e r d e t a i l .
H em atoxylin and e o sin s t a i n .
X l60.

P ig.

21.

U r e t e r from r a b b i t 6 w h ic h was n e c r o p s i e d f i v e
days a f t e r in o c u la tio n .
N e c r o s i s o f t h e m ucosa,
i n f i l t r a t i o n h y p s e u d o e o s i n o p h i l s , e dem a a n d
f i b r i n are i n evidence.
The l u m e n c o n t a i n s
b a c t e r i a , c e l l u l a r d e b r is , f i b r i n and p seu d o ­
eo sinophils.
G ra m -’V e i g e r t s t a i n .
X l6 o .

F ig.

22.

Focus of p s e u d o e o s in o p h ils i n and betw een th e
c o lle c tin g tubules.
R a b b i t 5» w h i c h was
n e c r o p s ie d f o u r days a f t e r i n o c u l a t i o n .
H e m a to : * y li n a nd e o s i n s t a i n .
X835*

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r

9k*

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V%

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#

■;» V

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B a c t e r i a , n e c r o t i c d e b r i s , pgeudoeos} n o p h i l s , and
c o lle c tin g tubuleg.
Figg. 9 a a i l6 c o n ta in t h i s
field .
G-ram-Waigert a t a i n .
X885-

F ig.

2^-.

C o l l e c t i n g t u b u l e s h o w i n g h y a l i n e "b od ie s i n t h e
cytoplasm of th e c e l l s .
R a b b i t 12 w h i c h was
n e c r o p s ie d se v e n te e n days a f t e r i n o c u l a t i o n .
G -ram -Weigert s t a i n .

5*
c

< Aj,y** »f . *■ f* *%, k

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25

Form ation of c r y s t a l s in n e c r o t ic m a te ria l a t the
edge of th e r e n a l p a p i l l a .
F i g s . 12, 15 a n d 20
contain th is f ie ld .
H e m a to x y lin and e o s i n s t a i n .
XS 8 5 .

-5 6 -

part

II.
PROPERTIES OE SOME COLONIAL PHASE
MARI ARTS OE CORY1TEBACTERTTTH RENALE

I n p r e v i o u s w o rk ( F e e n s t r a ,

19UU) i t h a d b e e n n o t e d t h a t t h e

c h a r a c t e r i s t i c s o f t h e Co r y n e b a c t e r l u m r e n a l e c u l t u r e s i s o l a t e d from
c a s e s o f "bovine p y e l o n e p h r i t i s w e r e , i n g e n e r a l ,

i d e n t i c a l w ith those

d e s c r i b e d "by B o r g e y ( 1 9 3 9 )* "but o f t e n p r o d u c e d c o l o n i e s w h i c h e x h i b i t e d
marked d i s s e m b l a n c e .
The t e n d e n c y o f d i p h t h e r o i d s t o v a r y i s w e l l known ( M e r c h a n t ,
1 9 3 5 )* b u t t h e c o l o n i a l p h a s e v a r i a t i o n o f _c. r e n a l e h a s r e c e i v e d s c a n t
a tte n tio n or p u b lic ity .
I n t h i s p r e l i m i n a r y w ork a s u r v e y o f t h e c o l o n i a l p r o p e r t i e s o f
t h e s t o c k c u l t u r e s o f C. r e n a l e was mads.

I n o c u l u m was s t r e a k e d c u t o r

t h i c k t r y p t o s e a g a r (D ifc o ) p l a t e s so t h a t s e p a r a t e c o l o n i e s co u ld
d e v e l o p t o m a t u r i t y ( 2 - 3 cm a p a r t ) .

C o l o n i e s w ere e x a m i n e d a s t o t e x t u r e

b y p r o b i n g a n d m o r p h o l o g i c a l l y u n d e r a d i s s e c t i n g m i c r o s c o p e by means o f
r e f l e c t e d l i g h t a n d a l s o b y o b l i q u e t r a n s m i t t e d l i g h t w h i c h was d i r e c t e d
t o w a r d t h e c o l o n i e s fr o m b e l o w a t a p p r o x i m a t e l y a U5 d e a r e e a n g l e by
means o f a m i r r o r a n d a m i c r o s c o p e lamp ( H u d d l e s o n , 19ll6 ) .
F i v e c u l t u r e s were ch o sen a s r e p r e s e n t i n g a t l e a s t t h r e e
d i f f e r e n t co lo n y ty p e s f o r f u r t h e r stu d y to determ ine th e biochem ical*
s e r o l o g i c a l and p a th o g e n ic p r o p e r t i e s and r e l a t i o n s h i p s .

The c o l o n i e s

w e r e r o u g h l y c l a s s i f i e d i n t o g r o u p s b y means o f t h e f o l l o w i n g k e y :
1.

Sm ooth,

s o f t p a s t y a n d no m a r k e d t e n d e n c y t o h o l d s h a p e when

probed.
a.

C i r c u l a r , e n t i r e , convex,
ground g l a s s a p p e a ra n c e .

c re am y w h i t e w i t h

b.

Irregular.

c.

Yellow t o orange c o l o r .

d.

Tend t o p r o d u c e f l a t s e c o n d a r y g r o w t h up on a g i n g .

-5 7 2.

O n ly s l i g h t l y p a s t y ,

marked te n d e n c y to h o l d shape and

f r a g m e n t when p r o b e d .

a.

C ircu lar, e n t i r e , convex, creamy w hite w ith
ground g l a s s appearance.

b.

Irregular.

c.

Yellow t o orange c o lo r .

d.

Tend t o p r o d u c e f l a t

s e c o n d a ry growth upon a g in g .

The f i v e c u l t u r e s c h o s e n f o r p r e l i m i n a r y s t u d y w e r e c l a s s i f i e d a s
follow s:
C u l t u r e ITo. 1
ii

m

ii

n

ii

it

I.e .

2

2. b.

3

2 . b.

ii

H

2 . a . d.

ii

5

2 . a . d.

A g g l u t i n a t i o n Re i s t i o n s h i p
A g g l u t i n a t i o n t e s t s w e r e c a r r i e d , o u t a s s u g g e s t e d i n t h e Manu al
o f M et ho ds f o r P u r e C u l t u r e S t u d y o f B a c t e r i a ( 1 9 ^ 5 ) w i t h t h e e x c e p t i o n
t h a t d i s t i l l e d w a t e r r a t h e r t h a n 0 . £f> p e r c e n t ITaCl s o l u t i o n was u s e d a s
t h e d i l u e n t a n d t e s t s w e r e i n c u b a t e d a t 3 7 ° f ° r 2U h o u r s .

A ntigens

w e r e p r e p a r e d "by g r o w i n g t h e o r g a n i s m s i n two p e r c e n t t r y p t o s e ( D i f c o ) ,
i n d i s t i l l e d w a t e r , a t a pH o f 7 . 5 f o r 2^ h o u r s a t 3 7 ° ^ . ;

centrifuging;

w a s h i n g o n c e i n O.B p e r c e n t p h e n o l s o l u t i o n and t h r e e t i m e s i n d i s t i l l e d
w ater; and a llo w in g to rem ain i n d i s t i l l e d w ater i n th e r e f r i g e r a t o r
u n t i l used.

A s t a b l e s u s p e n s i o n was o b t a i n e d by s u s p e n d i n g t h e o r g a n ­

i s m s i n d i s t i l l e d w a t e r , a l l o w i n g one h o u r f o r s e d i m e n t a t i o n o f c lu m ps
a nd r e m o v i n g a p o r t i o n o f t h e s u p e r n a t a n t f o r u s e .

These su sp e n sio n s

w e r e s t a n d a r d i z e d b y means o f a C e n c o - S h e a r d - S a n f o r d p h o t e l o m e t e r t o
a p p r o x i m a t e l y t h e d e n s i t y o f t u b e 0*5 o f M c F a r l a n d ’ s n e p h e l o m e t e r .

-<58-

A n tisera were prodxiced in rabbitB by s i x to ten intraven ous i n j e c t i o n s
of a heavy d i s t i l l e d water suspension of l i v i n g organisms at approxi­
mately weekly i n t e r v a l s .

'Results of two t r i a l s are given in Table I .
^able I

Antigens
1

2

3

1-1 2 8 0 *
1280

6ho
6ho

6U0
6ho

80

5120

80

1280

160
320

320
6ho

1280
320

320
ho

320

3

160
320

6U0
6ho

1280
1280

160
160

320
320

U

80
ho

so
160

loO

ho

80

ho

320
6ho

320
6ho

320
320

160
320

320
6ho

1

2

5
*

Anti sera

1st
2n d

h

5

80

JLWU

I n d i c a t e s h i g h e s t d i l u t i o n a t which co m p lete a g g l u t i n a t i o n to o k p l a c e .
S u s c e p t i b i l i t y t o t h e B a c t e r i c i d a l A c t i o n o f B o v i n e P la s m a
V a r y i n g num be rs o f o r g a n i s m s w e r e e x p o s e d t o a c o n s t a n t amount o f

b o v i n e p l a s m a i n a n a t t e m p t t o m e a s u r e a ny p o s s i b l e d i f f e r e n c e i n a n t i ­
g e n ic p r o p e r t i e s o r any d i f f e r e n c e i n v i r u l e n c e .

The t e s t s w e re s e t up

a s i n d i c a t e d i n t h e p r o t o c o l u s i n g 13 x 12*5 mm t u b e s and w e re i n c u b a t e d
i n a w a t e r b a t h a t 37°C.

The p l a s m a ( . 2 p e r c e n t , sodium c i t r a t e ) u s e d

was f r o m a n i n f e c t e d cow w h i c h h a d b e e n e x p o s e d i n t r a u r e t h r a l l y l h mo nth s
p r e v i o u s l y w i t h a c u l t u r e s u s p e n s i o n o f C5. r e n a l e .

The a n t i g e n s w ere

s t a n d a r d i z e d b y means o f a C e n c o - S h e a r d - S a n f o r d p h o t e l o m e t e r t o a p p r o x i ­
m a te t u b e 1 . 5 o f M c F a r l a n d 1 s n e p h e l o m e t e r .

S u b c u l t u r e s o f e a c h t u b o w er e

made a t i n t e r v a l s on t r y p t o s e a g a r p l a t e s b y s t r e a k i n g a l o o o f u l o f
i n o c u l u m on a n a r e a a p p r o x i m a t e l y 2 x 3
The r e s u l t s w e re n o t c o n c l u s i v e .

cra*
R e p e a t e d t e s t s showed t h a t

‘-5 9 c u l t u r e No.
resistan t

1 was m o s t s u s c e p t i b l e a n d t h a t c u l t u r e No.

to th e h a c t e r l e i d e l a c t i o n of b o v in e plasm a.

3

most

The o t h e r

t h r e e c u l t u r e s g a v e s o m ew ha t v a r i a b l e r e s u l t s r a n g i n g b e t w e e n t h e two
c u l t u r e s f i r s t m entioned.

Th e r e s u l t s

o f a ty p ic a l t r i a l a re given

i n Table I I .
P r o t o c o l of B a c t e r i c i d a l Test

1

Tube

2

D i s t i l l e d HaO

1

^

1

C u ltu re Susp.

0.5

Plasm a

0 ^ 3 ) l

D i lu tio n of
C ulture

1 -U

D ilu tio n of
Plasm a

0

I

]£

S

1

I

£

1

1

1

1

1

1

0.5

0.5

0.5

0.5

0.5

0 . 5 ^2 )

1

1

1

1

1

1

1-U

1 -1 2

I-3 6

108

32U

972

2916

1-2

1-2

1-2

1-2

1-2

1-2

1-2

1 ^

(1 )

A H q u a n t i t i e s g i v e n a r e i n ml .

(2.)

D i s c a r d 0 . 5 ml-

( 3)

0 . 5 ml b r o t h a d d e d t o t h e c o n t r o l t u b e .

•*•6o~

Ta b l e I I
R e s u lts of a T y p ica l T r i a l

C u ltu re

Time

1

1

0

2U
Ug
72
0

2

2U
us
3

u

1

6

++++<2 > ++++
+++ +
++++
++++
++++
++++
+

++++

+4-++
+++
2

++++

+++

I
++

4*4l
0

+
0
0

U7

3

+
5
1

+++ +
++++

++++

++++

++++

++++

++++
++++

++++

++++
++++

++++
++++

++++
++++

++++
++++

13

15

12

13

n

++
4-4*4"
4-4“
2

++
+-*-+
++
+++
+
+++
++
2

+++
5
n

g
I t

0

++++

++++

++++

++++

++++

+++

2U
Ug

++++
++++
++++

++++

++++
++++
+++

++++
4-4“4-44-4-4"

++++
+4-++
4*4-

++++
+++

++
+++
+++

u

lg

++++
•4-+++
++++
++++

"1—
r'r•T•
4■4■‘4*4"
4■4-

"iVfi i1'iI_
++++
++
12

-L._L.A
_I.
4*4"4"4

++++
++++

4-4-4-4+++ +

4-++
+++

20
7

lU
2

u

2
0

4"4-4—
f4-4*4-4
4-4-49

+++
++++
+

++
+++
++

++++
U

+

5

5

3

U

2U
Ug
72

0
2U
Ug
72

4-

1

++++

++++

++++

++++
++++
++++

+++ +
+++

++++
+++

++

+++

(1)

Time i n h o u r s .

(2)

I n d i c a t e s a m o un t o f g r o w t h ;
coloni es.

( 3)

tfumber o f c o l o n i e s p r e s e n t .

■

0
2

72

0

5

3

+++ +
++++

72

U

2

4-44-4
5

2

4-

U

( + ) i n d i c a t i n g a r r o r o x i m a t e l y 50

B iochem ical P r o p e r t i e s
G l u c o s e was f e r m e n t e d w i t h r e s u l t i n g a c i d p r o d u c t i o n "by a l l f i v e
cu ltu res,

hut

t h e a c t i o n o f c u l t u r e Ho. 1 w as somewhat w e a k e r t h a n t h a t

o f the o th e r fo u r c u ltu r e s .
A sim ilar s itu a tio n re su lted in
ten percent

s k im m i l k a e; a r ( t r y n t o s e ) ;

o th ers in halo production.

the study of c a s e in d ig e s tio n in
c u l t u r e ITo. 1 l e e r e d "behind t h e

-6 i-

P a th o g e n lc lty f o r R abbits
I n o r d e r to o b ta i n d a t a c o n ce rn in g th e d i f f e r e n t i a l v i r u l e n c e
of th e f i v e c u l t u r e s u n d e r s tu d y ,

a p p r o x i m a t e l y f o u r h i H i on o r g a n i s m s

( d i s t i l l e d w a t e r s u s p e n s i o n ) o f e a c h c u l t u r e grown on t r y n t o s e a g a r f o r
Rb h o u r s a t 37° C. w e r e i n j e c t e d i n t r a v e n o u s l y ,

e a c h i n t o two r a b b i t s .

D e t e r m i n a t i o n of t h e body w e i g h t s , hemoglobin l e v e l s ( Cenco-SheardS a n f o r d p h o t e l o m e + . e r , H of fm an m e th o d , l l U l ) ,

num be rs o f l e u k o c y t e s a n d

d i f f e r e n t i a l l e u k o c y t e c o u n t s w e r e made a t i n t e r v a l s .

The r a b b i t s w h i c h

d i d n o t d i e w e r e s a c r i f i c e d a t n i n e a nd t e n d a y s a f t e r i n o c u l a t i o n ,
n ecro p sied and the le s io n s recorded.

The l u n e ,

liv er,

spleen, h e a rt

b l o o d , b o n e m a rr o w , l e f t k i d n e y a n d u r i n a r y b l a d d e r w e r e c u l t u r e d on
tryotose agar.

T iss u e s f o r m icroscopic exam ination (lu n g , l i v e r ,

s p l e e n , k i d n e y , u r i n a r y b l a d d e r a n d u r e t e r s ) w e re f i x e d i n Z e n k e r 1 s
f l u i d a n d p a r a f f i n s e c t i o n s w e r e p r e p a r e d and s t a i n e d w i t h h e m a t o x y l i n
a n d e o s i n a n d by t h e G r a m - W e i g e r t m e t h o d f o r s t a i n i n g b a c t e r i a i n
sections.
A l l t h e r a b b i t s h a d grosG o r m i c r o s c o p i c l e s i o n s o f c o c c i d i o s i s
in the l i v e r .

T h i s may a l s o a c c o u n t f o r

i n i t i a l leu k o cy te counts o b tain ed .
rately .

some o f t h e h i g h e r t h a n a v e r a g e

O ther l e s i o n s w i l l be recorded sepa­

O c c a s i o n a l l y t h e l e s i o n s r e p o r t e d i n t h e g r o s 9 e x a m in a tio n were

m is s e d I n s e c t i o n i n g th e t i s s u e , b u t th e y were p r e s e n t i n a s u f f i c i e n t
number o f c a s e s t o a l l o w a d e q u a t e d e s c r i p t i o n .

R e p o r t s on I n d i v i d u a l

r a b b i t s follow .
Cases 1 and 2
R a b b i t s l a and l b were i n o c u l a t e d i n t r a v e n o u s l y w i t h c u l t u r e
No. 1.

None o f t h e o r g a n i s m s w e re r e c o v e r e d by c u l t u r e t e c h n i c a n d no

g r o s s o r m i c r o s c o p i c l e s i o n s were fo u n d .

What a p p e a r e d t o b e a clump o f

- 62 o i d s was s e e n i n t h e p e l v i s o f t h e r i g h t k i d n e y o f r a b b i t
Rabbit l a
Time^1 )

wbc^3 )

Hb ^

m ^)

L

P

B

S

* t <5)

0

12. U

1 3 ,6 0 0

3

60

35

1

1

1

1 1 .0

1 1 ,1 5 0

3

U6

UU

6

1

3

1 1 .0

I I .U 50

2

U9

37

9

3

2010

3

n .5

1 2 ,3 5 0

2

5?

3?

6

2

2022

7

u .5

1 5 ,^ 5 0

2

39

51

7

1

2002

9

11.2

11.850

3

62

29

6

0

2030

1763

202U
-

Rabbit lb
0

1 1 .6

12 ,750

2

62

31

u

1

1

li.l

5 ,8 5 0

1

H6

U2

9

2

3

10. U

13,850

u

53

33

7

3

19 lU

5

10. 8

1 0 ,9 0 0

2

52

38

6

2

1820

7

1 1 .9

16,350

U

U3

>+5

7

1

1S16

9

1 1 .3

12,050

3

55

35

6

1

IgUg

(1)

-

Time i n d a y s a f t e r i n o c u l a t i o n ; 0 i n d i c a , t i ng t h e

r to in o c u la tio n .

(2)

H e m o g l o b i n i n gra ms p e r 100 ml of b l o o d .

(3)

L e u k o c y t e s p e r c u b i c mm.

(k )

P ercent of mon oc ytes , lymphocytes, p s e u d o e o sin o p h ils,

b a s o p h ils and e o s in o p h ils .
(5)

Body w e i g h t i n gra m s .
Case 3

R a b b i t 2a was i n o c u l a t e d w i t h c u l t u r e No. 2.

O r g a n is m s w e r e

c u l t u r e d f r o m t h e p e l v i s o f t h e k i d n e y a n d fr o m t h e u r i n e .

The l u n g s

-6 3 -

w ore s l i g h t l y

congested.

The k id n e y s a p p e a r e d normal e x c e p t f o r t h e

p re se n c e o f a y e llo w c o n c r e tio n i n th e p e l v i s of th e l e f t kidney and a
s l i g h t p a p i l l a r y n e c r o s i s w h i c h was a l s o n o t i c e d i n t h e m i c r o s c o p i c
e x a m i n a t i on.

Hb

WBC

M

L

p

B

E

Wt.

0

12.1

9,700

0

63

3b

1

2

2050

1

12.1

7,950

o

U5

bj>

8

0

-

3

12.1

12,U00

H

52

b2

1

1

I 865

5

12.9

26,500

3

b6

30

17

U

1828

7

12.7

1 1 ,1 0 0

3

bs

U5

1

2

18 2U

9

1 2 .3

18,000

0

53

39

7

1

13 H6

Time

C ase ^
R a b b i t 2b was i n o c u l a t e d w i t h c u l t u r e TTo. 2 a n d d i e d on t h e
s e v e n th day.

Organisms w ere c u l t u r e d from t h e k id n e y p e l v i s and c o r t e x

an d from t h e u r i n e .

B oth k id n e y s were e n l a r g e d and e x h i b i t e d c o n g e s t i o n ,

c o r t i c a l ab sc e sses, p a u illa r y erosion,
m a te ria l i n th e pelves,
and congested.

a n d p u r u l e n t b l o o d y mucous

u r e t e r s and b la d d e r.

The u r e t e r s w e r e e n l a r g e d

Upon m i c r o s c o p i c e x a m i n a t i o n t h e l i v e r a n d s p l e e n

appeared congested.

The k i d n e y s showed edema, c o n g e s t i o n , h e m o r r h a g e ,

d e g e n e ra tio n of th e convoluted tu b u le s ,
th e t i p s of th e p a p i l l a e ,

and b & c te r ia l masses i n th e c o l l e c t i n g tu b u le s

a t the t i p s of the p a p illa e .
d e b r i s and d i p h t h e r o i d s .

a b sc e sses in the c o r tic e s and a t

The p e l v i s was f i l l e d w i t h b l o o d ,

The p e l v i c e p i t h e l i u m was n e c r o t i c and * h e

p e r i p e l v i c t i s s u e s were i n f i l t r a t e d w i t h f i b r i n ,
flu id .

tissu e

e r y th r o c y te s and serous

-614-

Hb

WBC

M

L

P

B

X

wt.

0

in . 5

£.550

2

75

19

3

1

2128

1

1 3 .3

3.700

1

U3

56

0

0

3

1 3 .7

7,950

7

>+9

U

2

0

19^3

5

1U.8

2 8 ,5 5 0

16

39

11

1

1310

Time

p

33

-

C as e 5
R a b b i t 3a was i n o c u l a t e d w i t h c u l t u r e No. 3*

O r g a n i s m s w e re

c u l t u r e d f r o m t h e k i d n e y p e l v i s and f r o m t h e "b la d d er .

The k i d n e y s w ere

s w o lle n and p a l e and e x h i b i t e d w h i t i s h s t r e a k s i n t h e m e d u lla r a d i a t i n g
tow ard th e c o r te x , p a p i l l a r y n e c r o s is ,
p e l v i c mucosa.

and yellow n e c r o t i c fo c i i n th e

The u r e t e r s w ere s l i g h t l y e n l a r g e d .

M icroscopically,

t h e k i d n e y s showed d e g e n e r a t i o n o f t h e c o n v o l u t e d t u b u l e s , p a p i l l a r y
e r o s i o n c h a r a c t e r i z e d by n e c r o s i s ,
eosinophils,

c o n g e s tio n , hemorrhage, pseudo­

and d i p h t h e r o i d s i n th e n e c r o t i c d e b r i s and i n s e v e r a l

c o lle c tin g tubules.

The p e l v i c mucosa was n e c r o t i c a n d t h e p e r i n e l v i e

t i s s u e was i n f i l t r a t e d w i t h s e r o u s f l u i d ,

Ti me

Hb

WBC

M

f i b r i n and p s e u d o e o s in o p h ils .

L

P

B

TP

Wt.

0

13.3

12,650

1

79

12

2

6

2320

1

12.1

5,3 5 0

0

30

60

10

0

-

3

10. U

9,350

9

6U

21

5

1

2332

5

10. U

9 ,6 5 0

7

6s

22

2

1

2288

7

1 3 .2

9 ,0 0 0

3

3?

62

1

2

225U

9

11.8

io,U 5o

3

66

30

1

0

2282

Cas e 6
R a b b i t 3b was i n o c u l a t e d w i t h c u l t u r e No • 3c u l t u r e d f r o m t h e k i d n e y p e l v i s a n d fr o m t h e b l a d d e r .

O rg a n is m s
No g r o s s o r m i c r o ­

s c o p i c a l t e r a t i o n s w e re a p p a r e n t b u t d i p h t h e r o i d s w e re s e e n i n t h e

-6 5 -

b l a d d e r , u r e t e r and k id n e y .

T he s e c n r a n i sr. s were v e r y l i k e l y d i f f e r e

f r o m t h e o n e s i n j e c t e d a 3 j u d g e d fr om t h e i r c u l t u r a l c h a r a c t e r i s t i c s .
Hb

W3C

y

L

P

B

•a

Tt.

0

11.7

? , 2 CO

3

70

2U

1

o

opUO

1

11. u

7,250

2

U7

50

0

1

X

1 1 .1

1 1 ,0 5 0

6

50

31

3

3

233U

5

1 1 .8

? , 300

x

S3

32

1

1

?xoo

7t

1 2 .1

? , 200

5

65

26

X

1

2U10

9

1 2 .3

300

6

59

XI

3

1

p s Uq

Ti me

tew

Case 7
Babb i t Ll*. wag i n o c u l a t e d . w i t h c u l t u r e Ho.
c u l t u r e d fr o m t h e <1 dney p e l v i s .

wi sf

ST: S "**c I*S

The k i d n e y s v e r ° n o r m a l i n s i z e a n d

c o lo r but th e l e f t kidney e x h ib ite d r a r i l l a r y n e c r o s is a n 4 abscess
f o r m a t i o n a nd t h e r i g h t k i d n e y e x h i b i t e d v e r y s l i g h t u a p i l l a r y n e c r o s i s .
S e c t i o n s d i d n o t show t h e t i n s of t h e r e n a l n a v i 11a e , h u t t h e r e l v i c
e r o i t h e l i u a showed s l i g h t
c o n t a i n e d some f i b r i n .

Time

Hb

d e g e n e r a t i v e c h a n g e s and t h e p e r i p e l v i c t i s s u e s
T h e r e was n e c r o t i c t i s s u e d e b r i s i n t h e o e l v i s .
7T3C

M

L

p

3

3

Wt.

0

'ib.i

1 2 ,1 0 0

1

*3

10

3

3

2266

1

11. S

3.550

0

T7
>'

61

1

1

-

X

1 1. u

2 2 ,3 0 0

r
D

70

PX
*—^

0

1

2111

5

1 2 .5

1 1 ,6 5 0

5

7U

15

5

1

20X6

7

1 2 .9

1 6 , bnQ

X

61

22

lb

C

20X0

c>

1 3 .2

10,700

0

6c

XO

0

2176

•3

Case 5
B a b b i t hb wa3 i n o c u l a t e d w i t h c ti —t)u. r e 'To

Ho o r g a n !

-6 6 -

c u l t u r e d from t h i s r a h h i t .
tio n of the co rtex ,
the p a p illa .

The r i g h t k i d n e y e x h i b i t e d a r>atchy c o n g e s ­

a sm all w hite fo cu s i n th e m edulla and c o n g e stio n of

I n s t a i n e d s e c t i o n s t h e r i g h t k i d n e y showed a s m a l l f i b r o t i c

nodule i n t h e in n e r m edulla and th e e p ith e liu m of th e c o l l e c t i n g tu b u le s
i n the m edulla of th e l e f t kidney appeared s l i g h t l y d eg en erated .
Hb

TOC

M

L

P

B

s

wt.

0

1 2 .1

9.850

0

60

36

2

2

2U00

1

12.5

6 ,5 0 0

2

30

66

1

1

3

1 3 .1

8 ,0 0 0

6

56

32

3

3

2396

5

13.5

U .7 5 0

1

51

Hi

6

1

2308

7

13.?

3,250

1

55

36

7

1

2312

9

13.7

1 0 ,1 0 0

2

U5

Us

U

l

2UU0

Time

Case 9
R a b b i t 5 a was i n o c u l a t e d w i t h c u l t u r e No. 5*
c u l t u r e d from t h i s r a b b i t .
sections.

No o r g a n i s m s w e r e

The o n l y l e s i o n s f o u n d w e r e i n t h e s t a i n e d

T h e r e was a s l i g h t i n c r e a s e o f p s e u d o e o s i n o p h i l s i n t h e l u n g ,

and a s l i g h t i n c r e a s e o f p h a g o c y to s is of b lo o d pigm ent i n th e sp le e n .
Hb

TOC

M

L

P

B

E

TTt.

0

11.8

3 ,9 5 0

2

65

30

2

1

2152

1

11.8

7 ,8 0 0

0

59

Uo

1

0

-

3

11.8

10,700

5

53

39

2

3

2162

5

1 2 .3

1 2 .7 5 0

U

62

28

5

1

2120

7

1U.6

16,800

u

59

3^

l

2

2182

9

12. k

9 ,2 5 0

5

US

HU

3

0

223s

Time

C ase 10
R a b b i t 5b was i n o c u l a t e d w i t h c u l t u r e Ho. 5-

No o r g a n i s m s w e re

-6 7 c u l t u r e d fr o m t h i s r a b b i t .
Time

No f r r o s s o r m i c r o s c o p i c l e s i o n s w e r e a p p a r e n t .

Hb

WBC

M

L

p

B

E

w t.

0

11. U

9 ,2 5 0

1

76

13

3

2

2300

1

li.i

5 , Uoo

0

UU

50

5

1

-

3

1 1 .6

1 3 .7 5 0

2

Ug

U5

5

0

2230

5

1 2 .5

9 ,8 0 0

3

56

31

g

2

2282

7

1 2 .9

1 1 ,0 5 0

3

U9

U2

5

1

2335

9

12.U

1 0 ,^ 50

1

5U

Ul

2

p

2UUU

Table I I I
Summary o f D a t a
TCidney^1 ^
( .
Lesions C ulture
'

Wt. L o s s

Wt. o t e n d
Comp, t o Beedn

Max.# i n c . ^ ^
i n WBC

C ulture

R abbit

1

la

-

-

22

+6

13

1

lb

-

-

Rone

+95

28

2

2a

+

+

225

- 20U

156

2

2b

+

+

uoo

-Uoo

23U

3

3a

+

+

66

3

313

-

U

Ua

+

+

U

Ub

H
r

-

92

5

5a

-

-

32

+86

65

5

5b

-

-

20

+lUU

U9

-3 3

drop

None

+300

35

233

-9 0

su

+Uo

3

(1)

M icro sco n ic or erross l e s i o n s in the k i dneys.

(2)

R e c o v e r y o f t h e o r i g i n a l o r g a n i s m "by c u l t u r e a t n e c r o p s y .

(3)

L o w e s t w e i g h t r e c o r d e d d u r i n g e x p e r i m e n t co m pa re d to wele-ht a t
t h e b e g i n n i n g (G r a m s ) .

(U)

W e i g h t a t t h e end o f t h e e x p e r i m e n t compared t o t h e w e i g h t a t
b e g i n n i n g (Grams).

(5 )

The maximum p e r c e n t i n c r e a s e i n t h e l e u k o c y t e c o u n t .

-6 « D lscusslon
The r e s u l t s o f t h i s p r e l i m i n a r y work i n d i c a t e t h a t t h e r e w a r e
m a rk e d d i f f e r e n c e s i n t h e m o r p h o l o g y a n d p r o p e r t i e s o f t h e c o l o n i e s
c h o s e n f o r s t u d y from s t o c k c u l t u r e s w h i c h w e r e o r i g i n a l l y i s o l a t e d fr om
c a t t l e having p y e lo n e p h r itis .

T h a t a l l t h e s e v a r i a t i o n s w e re p r e s e n t i n

t h e c u l t u r e s a a o r i g i n a l l y i s o l a t e d i a v e r y d o u b t f u l and c o u l d n o t h e
m aintained.
As may "be n o t e d "by e x a m i n a t i o n o f t h e p h o t o g r a p h s and t h e
c l a s s i f i c a t i o n of th e c o lo n ie s ,

the g r e a t e s t m orphological d if f e r e n c e s

w e r e b e t w e e n c u l t u r e No. 1 and t h e o t h e r f o u r c u l t u r e s .

The m o r p h o l o g y

o f c u l t u r e s No. 2 a nd No. 3

th a t of c u l t u r e s

somewhat s i m i l a r a n d a l s o

No. U a n d No. 5»
The s e r o l o g i c a l r e a c t i o n s o f Q o r y n e b a c t e r i u m r e n a l e a r e a t b e s t
no ne t o o s a t i s f a c t o r y .

The r e s u l t s o f t h e a g g l u t i n a t i o n t e s t s show t h a t

t h e r e a r e a n t i g e n i c d i f f e r e n c e s and s i m i l a r i t i e s b e t w e e n t h e d i f f e r e n t
c u ltu re s but there is l i t t l e

o r no c o r r e l a t i o n w i t h t h e m o r p h o l o g i c a l

c h a r a c t e r ! s t i e s of the c o lo n ie s .
The r e s u l t s o f t h e b a c t e r i c i d a l r e a c t i o n s shew a f a i r l y c l o s e
c o r r e l a t i o n w ith the m orphological c h a r a c t e r i s t i c s .
t h e most s u s c e p t i b l e ,

C u l t u r e No. 1 was

c u l t u r e e No. 2 a n d No. 3 were t h e l e a s t s u s c e p t i b l e

a n d t h e r e a c t i o n s o f c u l t u r e s No. H a n d No. 5 w e re i n t e r m e d i a t e .

The

plasma, u s e d i n t h i s c a s e v e r y l i k e l y c o n t a i n e d some s p e c i f i c a n t i b o d i e s
b e c a u se t h i s anim al had been exposed, i n t r a u r a t h r a . l l y ,
a n d was s t i l l p a s s i n g o r g a n i s m s i n t h e u r i n e .
here,

I 1' mo nth s b e f o r e

Other t e s t s , not r e c o r d e d

showed t h a t t h e p l a s m a from a cow n o t p a s s i n g o r g a n i s m s was l e s s

p o ten t in actio n .
The p a t h o g e n i c i t y t e s t , u s i n g r a b b i t s , was o f c o u r s e w o e f u l l y
i n a d e q u a t e b e c a u s e so fe w a n i m a l s w e re u s e d t o t e s t e a c h c u l t u r e .

The

-nQ-

pflsnj.its ,

s s s u m m e r iz e d i n ma b l e I I I ,

p a th o g e n i c ity of the c u ltu r e ? .

wo u ld i n d i c a t e ma rkeo d i f f e r e n c e s i n

The b a c t e r i o l o g y , p a t h o l o g y ,

end h e m a t o l o g y

of t h e i n f e c t e d r a b b i t s w i l l n o t h e d.i s c u s s e d h e r e b e c a u s e t h e s e s u b j e c t s
w ere c o v e r e d c o r e t h o r o u g h l y i n t h e f i r s t n a r t of t h i s

thesis (Part I).

The w e i g h t s o f t h p s e r a h h i t s w e re r e c o r d e d a t i n t e r v a l s e ^ t e r n o t i c i n g i n
p r e v i o u s work t h a t i n f e c t e d r a h h i t s l o s t w e i g h t during* t h e c o u r s e o f t h e
in fectio n .

In general,

th e r a h h i t s l o s t w eight a f t e r i n o c u l a t i o n and t h e

i n f e c t e d ones c o n t i n u e d to l o s a w eight w hereas th e ones which d id not
become i n f e c t e d grained w e i g h t a f t e r t h e i n i t 1**! s l i g h t l o s s .
I t i s a p p a r e n t t h a t t h e c u l t u r e s w h i c h would l i k e l y h Q c l a s s e d a s
" r o u g h " , m o r p h o l o g i c a l l y , w e r e more p a t h o g e n i c f o r r a h h i t s a n d l e s s s u s ­
c e p t i b l e t o t h e h a s t e r i c l d e l a e t i on o f "bovine p l a s m a a n 4 t h e c u l t u r e t h a t
would most l i k e l y h e c l a s s e d a s "s m oo th " was m a r k e d l y l e g s p a t h o g e n i c f o r
r a h h i t s a n d s o r e s u s c e p t i b l e t o t h e " b a c t e r i c i d a l a c t i o n o f "bovine p l a s m a .
I n t h e p a s t , s u c h work h a s b e e n done on t h e r e l a t i v e v i r u l ° n c e of
b acterial varian ts.
the l i t e r a t u r e

I t i s n o t c o n s i d e r e d d e s i r a b l e a t t h i s time t o rev ie w

on t h i s s u b j e c t .

Sample r e f e r e n c e s t o s u c h work would

i n c l u d e D e Z r u i f ( 1 9 5 1 ) , H u d d l e s o n e t a l ( l ? - - f ) , a nd I r w i n and T e a c h ( 1 9 ^ 9 ) .

i. b r i e f p r e l i m i n a r y s t u d y and d i s c u s s i o n o f t h e m o r p h o l o g y ,
agglutination re la tio n sh ip s,

r e a c t i o n to the b a c t e r i c i d a l a c t i o n of

b o v i n e plasm®., and t h e p a t h o g e n i c i t y o ' s o r e o f t h e c o l o n y t y p e s f o u n d
ir. s t o c k c u l t u r e s o f T c r m e h a c t e r i u r r e n a l e have b e e n p r e s e n t e d .
At p r e s e n t ,

i t may be c o n c l u d e d t h a t t h e " r o u g h " t y p e of c u l t u r e

i s more o s t h c g e n i c f o r r a b b i t s a n d more r e s i s t a n t
a c t i o n of b o v i n e p l a s m a t h a n t h e smooth t y p e .

to t h e b a c t e r i c i d a l

-7 0 -

Ref p r i n c e s
B e r ^ e y , D. H. , e t a l .
1-979 Manu al o f d e t e r m i n a t i v e b e c t e r i o l o a y .
5 t h ed.
W i l l i a m s a nd A l i k i n a Co. , B a l t i m o r e , Md.
D e K r u i f , P . H.
1921 D i s s o c i a t i o n o f m i c r o b i c s n e c i e s . I . C o e x i s t e n c e
of i n d i v i d u a l s of d i f f e r e n t d e g re es of v i r u l e n c e i n c u l t u r e s of
t h e " b a c i l l u s of* r a b b i t s e p t i c e m i a .
J . Exp. M ed ., 7 3» 77 3 “ 7 ^ 9 .
P e e n s t r a , E. S.
19 UU S t u d i e s on t h e e t i o l o g y a n d - p a t h o l o g y o f
i n f e c t i o u s Bovine p y e l o n e p h r i t i s .
T hesis.
School of Graduate
Studies.
M ichigan S t a t e C o lle g e .
H o f f m a n , W. S* 15b-l P h o t e l o m e t r i c c l i n i c a l c h e m i s t r y .
Morrow and C o . , N.Y.

W illiam

H u d d l e a o n , I . P . , Wood, E. P . , G ro s s m a n , A. P. and B e n n e t t , G. P..
IQb9 The " b s . c t e r i c i d a l a c t i o n o f "bovine "blood f o r "Brucella, a n d
i t s p o s s i b l e sigrnif ic s.n c e . J . B a c t . ,
261-277.
H uddleson, I . F.
1GU6 The mucoid -p ha se s o f t h e grenus b r u c e l l a .
Am. J . V e t . R e s . , 7 , 5 - 1 0 .
I r w i n , M. R. and. B e a c h , B. A.
19^6 D i f f e r e n t i a l " b a c t e r i c i d a l
a c t i v i t y o f b o v i n e serum toward, s t r a i n s o f B r u c e l l a , ab o r t u s o f
h ig h and low v i r u l e n c e .
J . A e r . R e s . , 7 2, K 9 - 9 i .
Manual of m e th o d s f o r p u r e c u l t u r e s tu d y o f b a c t e r i a .
1979
P u b l i c a t i o n s , G e n e v a , 1T.Y.
(C urrent re v is io n s used).
M e r c h a n t , I . A.
1935
dom estic a n im a ls.

B iotech

C o r y n e b a c t e r i a a s s o c i a t e d , w i t h d i s e a s e s of
J . B a c t . , 3(~'« 9 5 - 1 1 2 .

Fig.

k

26.

C o l o n y o f c u l t u r e No. 1.
I'wo w e ek s e-rowth on
t r y p t o s e a g a r a t roor. t e r r o e r a t u r e a f t e r t h e
f i r s t day a t 1 7 ° C. X10.
<The c o l o n i e s d e p i c t e d i n F i£ - s . 9 7 , off, ?Q
a n d 3f we r p grown u n d ^ r s i m i l a r c o n d i t i o n s ) .

P ig.

27.

C o l o n y o f c u l t u r e No. 2.

X10.

F ig.

2 8.

C o lo n y o f c u l t u r e No. 3 .

X 10.

m ....

list!

P ig.

&

29.

C ol o n y o f c u l t u r e No. U«

X10.

Fig.

30*

C o lo n y o f c u l t u r f ? No. 5»

X 10.