survival of vibrio cholerae and
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8/10/2019 Survival of Vibrio Cholerae And
1/8
1982, 43(3):578.Appl. Environ. Microbiol.
M A Hood and G E Nesswaters and sediments.
Escherichia coli in estuarineSurvival of Vibrio cholerae and
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8/10/2019 Survival of Vibrio Cholerae And
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V o l .
4 3 ,
N o .
3
P P L I E D
AND
ENVIRONMENTAL
M I C R O B I O L O G Y ,
M a r . 1 9 8 2 ,
p .
5 7 8 - 5 8 4
0 0 9 9 - 2 2 4 0 / 8 2 / 0 3 0 5 7 8 - 0 7 0 2 . 0 0 / 0
S u r v i v a l o f
V i b r i o c h o l e r a e a n d E s c h e r i c h i a
c o l i i n
E s t u a r i n e
W a t e r s
a n d
S e d i m e n t s
MARY A . HOOD* A N D
G R E G O R Y E . N E S S
D e p a r t m e n t o f
B i o l o g y ,
T h e
U n i v e r s i t y
o f
W e s t
F l o r i d a ,
P e n s a c o l a ,
F l o r i d a
3 2 5 0 4
R e c e i v e d
1 2
A u g u s t
1 9 8 1 / A c c e p t e d
3 0 N o v e m b e r 1 9 8 1
I n i n
v i t r o e s t u a r i n e
w a t e r
a n d
s e d i m e n t c h a m b e r s ,
t h e
s u r v i v a l o f
V i b r i o
c h o l e r a e a n d
E s c h e r i c h i a c o l i w a s d e t e r m i n e d
b y
p l a t e
c o u n t i n g
a n d
d i r e c t
c o u n t i n g
t e c h n i q u e s .
V . c h o l e r a e s t r a i n s
i n c l u d e d
e n v i r o n m e n t a l ,
c l i n i c a l ,
a n d
s e r o t y p e
0 1 a n d
n o n - O 1
i s o l a t e s ,
w h e r e a s
E . c o l i
s t r a i n s
i n c l u d e d
A T C C 2 5 9 2 2
a n d
a
f r e s h l y c u l t u r e d human
i s o l a t e . R e c o v e r y o f V . c h o l e r a e
v a r i e d s i g n i f i c a n t l y
w i t h
i n c u b a t i o n
t e m p e r a t u r e .
G r o w t h
a n d e x t e n d e d p e r i o d s
o f
s u r v i v a l
o c c u r r e d
i n s t e r i l e
s e d i m e n t s ,
s t e r i l e
w a t e r s ,
a n d n o n s t e r i l e
w a t e r s ,
b u t
n o t
i n n o n s t e r i l e
s e d i m e n t s .
I n c o nt r a s t t o
V .
c h o l e r a e ,
v i a b l e
c e l l s
o f
E .
c o l i
d e c r e a s e d
r a p i d l y
i n
b o t h s t e r i l e a n d n o n s t e r i l e e s t u a r i n e
w a t e r s .
D i r e c t c o u n t s
r e v e a l e d
t h a t E .
c o l i
c e l l s w e r e i n t a c t
i n t h e e s t u a r i n e
w a t e r ,
b u t
a t t e m p t s
t o
r e s u s c i t a t e
t h e m
w e r e
u n s u c c e s s f u l .
T h e
d a t a
s u g g e s t
t h a t
V .
c h o l e r a e s u r v i v e s
b e t t e r
i n
e s t u a r i n e
w a t e r s
t h a n
E . c o l i . T h e r e s u l t s
may e x p l a i n
t h e r e c e n t o b s e r v a t i o n s
t h a t V .
c h o l e r a e
l e v e l s d o
n o t c o r r e l a t e
w e l l
w i t h f e c a l c o l i f o r m
c o n c e n t r a t i o n s
i n
e s t u a r i n e w a t e r s .
F u r t h e r m o r e ,
t h e
r e s u l t s
a d d
i n c r e a s i n g
e v i d e n c e
t o
s u p p o r t
t h e
t h e o r y
t h a t V .
c h o l e r a e
i s
a n a u t o c h t h o n o u s
b a c t e r i u m
i n e s t u a r i e s .
T h e
common
o c c u r r e n c e o f
V i b r i o
c h o l e r a e
i n
t h e
e s t u a r y
( 7 , 8 , 1 3 , 1 4 ,
1 8 )
h a s l e d
t o
t h e
c o n c l u s i o n t h a t t h e
o r g a n i s m
i s
a n
i n d i g e n o u s
r e s i d e n t
o f
t h e
e s t u a r y 8 ,
1 4 ) .
F u r t h e r m o r e ,
t h e r e i s
i n c r e a s i n g
e v i d e n c e
s u g g e s t i n g
t h a t t h e
p r e s e n c e
o f V . c h o l e r a e
d o e s n o t
c o r r e l a t e
w e l l
w i t h l e v e l s
o f f e c a l c o l i f o r m s
( 8 ,
1 3 ,
1 4 ) .
T h e
p o s s i b i l i t y
t h a t
f e c a l c o l i f o r m s
ma y
n o t i n d i c a t e
t h e p r e s e n c e
o f V . c h o l e r a e
i n e s t u a r i n e
w a t e r s
i s
e s p e c i a l l y s i g n i f i c a n t f r o m
a
p u b l i c
h e a l t h
s t a n d p o i n t ,
s i n c e
f e c a l c o l i f o r m s h a v e h i s t o r i c a l -
l y
b e e n
u s e d a s
i n d i c a t o r s o f w a t e r a n d s h e l l f i s h
q u a l i t y .
W i t h t h e r e c e n t o u t b r e a k s
o f c h o l e r a
a n d
g a s t r o i n t e s t i n a l
d i s o r d e r s c a u s e d
b y
V . c h o -
l e r a e
2 , 4 - 6 , 2 0 )
a n d
t r a c e d t o
s h e l l f i s h ,
t h e
q u e s t i o n
b e c o m e s
e v e n
m o r e
r e l e v a n t .
T h e r e h a v e b e e n
many
s t u d i e s
o n t h e s u r v i v a l
o f
t h e c o l i f o r m E s c h e r i c h i a
c o l i
i n
s e a w a t e r
( 1 0 ,
1 9 )
a n d
a
n u m b e r o f
e a r l y
w o r k s o n
t h e
s u r v i v a l
o f
V .
c h o l e r a e
i n
s e a w a t e r a s
r e v i e w e d
b y P o l -
l i t z e r
( 1 6 ) .
S u r v i v a l
o f
c o l i f o r m s i n
m a r i n e
w a -
t e r s
w a s
c o n c l u d e d
t o b e
a f u n c t i o n o f
s a l t
c o n c e n t r a t i o n , p r e d a t i o n ,
c o m p e t i t i o n b y
n a t i v e
m i c r o f l o r a ,
h e a v y
m e t a l s , a n d
n u t r i e n t s ( 3 , 1 1 ,
1 5 ) .
V .
c h o l e r a e
s u r v i v a l
i n s e a w a t e r w a s f o u n d
t o
b e
r e l a t e d
t o t e m p e r a t u r e , p H , s a l t c o n c e n t r a -
t i o n ,
t h e
a m o u n t
o f o r g a n i c
m a t e r i a l , a n d t h e
d e g r e e o f
b a c t e r i a l
c o n t a m i n a t i o n ( 1 6 ) .
A l t h o u g h
t h e s e s t u d i e s
h a v e
r e p o r t e d
t h e
s u r v i v a l
r a t e s
o f
E .
c o l i a n d V .
c h o l e r a e , t h e r e i s
n o c o m p a r a t i v e
i n f o r m a t i o n
o n t h e s u r v i v a l o f t h e s e
t w o
o r g a n -
i s m s
i n
e s t u a r i n e
e n v i r o n m e n t s .
T h u s , i t w a s t h e
p u r p o s e
o f t h i s
s t u d y
t o e x a m i n e
t h e s u r v i v a l
o f
s t r a i n s
o f
V . c h o l e r a e i n e s t u a r i n e w a t e r s a n d
s e d i m e n t s
a n d
t o
c o m p a r e
t h e i r
s u r v i v a l
t o
t h a t
o f E s c h e r i c h i a
c o l i .
M A T E R I A L S A N D M E T H O D S
S u r v i v a l
c h a m b e r s . W a t e r
a n d
s e d i m e n t
w e r e
c o l -
l e c t e d
f r o m
A p a l a c h i c o l a B a y , F l a . ,
w i t h a
P o n a r
g r a b
( W i l d l i f e
S u p p l y
C o . ,
S a g i n a w ,
M i c h . )
a n d
s t e r i l e
w a t e r
b o t t l e s . T h e s e d i m e n t s a m p l e s
w e r e p l a c e d
o n
i c e ,
r e t u r n e d t o t h e
l a b o r a t o r y ,
a n d
k e p t
a t
4 C
u n t i l
t h e
s u r v i v a l c h a m b e r s
w e r e
e s t a b l i s h e d
( w i t h i n
5
d a y s
a f t e r
c o l l e c t i o n ) .
W a t e r
s a m p l e s
w e r e
s t o r e d
a t
a m b i -
e n t
t e m p e r a t u r e
a n d f i l t e r e d
t h r o u g h
a
Whatman
n o . 1
f i l t e r
t o
r e m o v e
l a r g e
d e t r i t a l
p a r t i c l e s .
S a l i n i t i e s o f
t h e
w a t e r s r a n g e d f r o m 6 t o
2 0 I / o I .
I n t o
s t e r i l e
g l a s s
c o n t a i n e r s
( 2 0 0
m l ) d e s i g n a t e d
t h e
s u r v i v a l
c h a m b e r s ,
1 0 0
m l
o f
t h e
c o l l e c t e d
e s t u a r i n e
w a t e r
w a s a d d e d .
S e d i m e n t c h a m be r s
w e r e p r e p a r e d
w i t h 5 0 g o f s e d i m e n t
a n d
5 0 m l
o f
e s t u a r i n e
w a t e r . T h e
s t e r i l e
w a t e r a n d s e d i m e n t
c h a m b e r s w e r e a u t o c l a v e d
a t s t a n d a r d
c o n d i t i o n s .
A l l
c h a m b e r s w e r e
a l l o w e d
t o
e q u i l i b r a t e
a t
2 0 C
f o r 4 8
h
b e f o r e
i n o c u l a t i o n ,
a n d t w o
o r
t h r e e
r e p l i c a c h a m b e r s w e r e
p r e p a r e d
f o r
e a c h
e x p e r i m e n t .
C u l t u r e
p r e p a r a t i o n .
E n v i r o n m e n t a l
V .
c h o l e r a e
s t r a i n s
w e r e i s o l a t e d
f r o m
o y s t e r s
c o l l e c t e d
f r o m A p a -
l a c h i c o l a
B a y
b y
t h e
m e t h o d s
o f
Hood
e t a l .
( 1 3 ) a n d
K a p e r e t a l .
( 1 4 )
a n d
w e r e m a i n t a i n e d o n T r y p t i c a s e
s o y
(BBL M i c r o b i o l o g y S y s t e m s ,
C o c k e y s v i l l e , M d . ) -
1
N a C l
a g a r .
V .
c h o l e r a e
s t r a i n s
i n c l u d e d a n
e n v i -
r o n m e n t a l
0 1
I n a b a ,
W F / 1
( 1 3 ) ,
a n
e s t u a r i n e
w a t e r -
a d a p t e d
0 1
s t r a i n ,
f i v e
e n v i r o n m e n t a l
n o n - O 1 s t r a i n s ,
W F / 2 3 , W F / 5 4 ,
W F / 1 0 3 ,
W F / 1 0 8 ,
a n d
W F / 1 1 1 ,
a n d a
c l i n i c a l
0 1
s t r a i n
( 6 ) .
N o n - O 1
V .
c h o l e r a e s t r a i n s w e r e
t y p e d b y
Ron
S i e b e l i n g ,
L o u i s i a n a S t a t e
U n i v e r s i t y ,
B a t o n
R o u g e , a s
T T
( W F / 2 3 ) , Un k ( W F / 5 4 , W F / 1 0 8 ,
5 7 8
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8/10/2019 Survival of Vibrio Cholerae And
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V . C H O L E R A E
AND
E .
COLI I N
E S T U A R I E S
5 7 9
o
7
10
10 5
1 0 4
1 0 2
1 0 1
1
2 3
4
5
6 7
8 9
1 0 1 n 1 2
Days
F I G . 1 . E f f e c t
o f i n o c u l u m
s i z e on t h e s u r v i v a l o f
V .
c h o l e r a e
0 1
( W F / 1 ) i n e s t u a r i n e
w a t e r s a n d
s e d i -
m e n t s i n c u b a t e d a t 2 0 C . T h e v a l u e s
a re
e x p r e s s e d a s
c o l o n y - f o r m i n g
u n i t s .
a n d
W F / 1 0 3 ) ,
a n d
B B
( W F / 1 1 1 ) . T h e e s t u a r y - a d a p t e d
i s o l a t e
was s t r a i n
W F / 1
p l a c e d
i n
e s t u a r i n e
w a t e r s .
T h e
o r g a n i s m
was
r e c o v e r e d
f r o m
t h e
w a t e r
a f t e r
6
w e e k s a n d
r e v i t a l i z e d
o n
T r y p t i c a s e s oy b r o t h
( B B L ) .
T h e
E .
c o l i
s t r a i n s
i n c l u d e d
A TCC
2 5 9 2 2
a n d a
f r e s h l y
i s o l a t e d
s t r a i n f r o m human
f e c e s .
E .
c o l i s t r a i n s
were
b i o c h e m i c a l l y
c o n f i r m e d
w i t h
t h e A P I 2 0 E
s y s t e m
( A n a l y t a b
P r o d u c t s , P l a i n v i e w ,
N . Y . ) .
V .
c h o l e r a e
a n d
E .
c o l i
i s o l a t e s were
grown
i n
T r y p t i c a s e
s o y
b r o t h - 1
N a C l
f o r 2 4 h
a t
3 5 C .
C u l t u r e s
were
c e n t r i f u g e d
a t
2 0 0 0 rpm f o r 2 0 m i n a n d
w a s h e d
i n
p h o s p h a t e - b u f f e r e d s a l i n e . T h e
p r o c e d u r e
wa s r e p e a t e d
t w i c e ,
a n d t h e
c e l l s were
i n o c u l a t e d
i n t o
t h e s u r v i v a l
c h a m b e r s .
I n c u b a t i o n t e m p e r a t u r e s , i n i -
t i a l i n o c u l u m
s i z e ,
a n d
t i m e a n d
l o c a t i o n o f w a t e r a n d
s e d i m e n t
c o l l e c t i o n were
v a r i e d .
R e c o v e r y
o f
v i a b l e
c e l l s .
A t
s e l e c t e d t i m e
i n t e r v a l s ,
s a m p l e s
were
t a k e n f r o m t h e c h a m b e r s
a n d
d i l u t e d i n
p h o s p h a t e - b u f f e r e d
s a l i n e b l a n k s . E n u m e r a t i o n was
p e r f o r m e d b y
t h e
s t a n d a r d
p l a t e
c o u n t m e t h o d o n
b r a i n
h e a r t
i n f u s i o n
agar ( D i f c o
L a b o r a t o r i e s , D e t r o i t ,
M i c h . ) , T r y p t i c a s e
s o y - 1
NaCl
a g a r ,
a n d
t h i o s u l f a t e
c i t r a t e b i l e s a l t
sucrose
agar ( D i f c o )
f o r
V .
c h o l e r a e .
F o r E .
c o l i
recovery,
e o s i n
m e t h y l e n e
b l u e
( E M B )
agar ( D i f c o )
a n d b r a i n h e a r t
i n f u s i o n agar
were u s e d .
E n u m e r a t i o n
o f
V .
c h o l e r a e a n d
E .
c o l i
c o l o n i e s
was
p e r f o r m e d
b y
c o m p a r i n g
t h e m w i t h
c o n t r o l c o l o n i e s .
01
s t r a i n s
were
c o n f i r m e d
b y u s i n g p o l y v a l e n t
a n t i s e -
rum
( D i f c o ) , w h e r e a s n o n - O 1 s e r o t y p e
a n d E .
c o l i
c o l o n i e s were
c o n f i r m e d
b y
u s i n g
t h e
API 2 0 E
S y s -
t e m .
I n
a d d i t i o n ,
t h e
most
p r o b a b l e
n u m b e r
f o r f e c a l
c o l i f o r m s was d e t e r m i n e d
b y u s i n g
t h e
s t a n d a r d m e t h -
o d w i t h
l a u r y l
t r y p t o s e
b r o t h
( D i f c o )
a n d
i n c u b a t i o n
a t
3 5 C
f o r 4 8
h
1 .
P o s i t i v e
t u b e s
were
t r a n s f e r r e d
t o E C
b r o t h
( D i f c o )
a n d
i n c u b a t e d
a t
4 4 . 5 C
f o r 4 8
h .
P o s i t i v e
t u b e s were t h e n s t r e a k e d o n t o EMB
a g a r . A t t e m p t s
w e r e m a d e
t o r e s u s c i t a t e
t h e E . c o l i c e l l s i n t o
l a c t o s e
b r o t h ( D i f c o ) b y i n c u b a t i n g
f o r
3
h a t
2 0 C
b e f o r e
i n c u b a t i o n f o r 4 5 h
a t
3 5 C .
T u b e s w h i c h s h o w e d
g r o w t h
w i t h
o r w i t h o u t g a s w e r e s t r e a k e d o n t o l a c t o s e
a g a r
a n d E M B a g a r
a n d w e r e t r a n s f e r r e d
t o E C b r o t h
a n d
i n c u b a t e d
a t
4 4 . 5 C
f o r
4 8
h .
E C
t u b e s w i t h g a s
o r
g r o w t h
w e r e s t r e a k e d
o n t o
l a c t o s e
a n d EMB
a g a r .
C o l o n i e s w e r e c o n f i r m e d o n
t h e A P I
2 0 E s y s t e m .
D i r e c t
c o u n t s .
C e l l s w e r e e n u m e r a t e d
b y
t h e
a c r i d i n e
o r a n g e
d i r e c t
c o u n t i n g p r o c e d u r e
o f H o b b i e e t a l .
( 1 2 ) .
A Z e i s s
s t a n d a r d
1 8 m i c r o s c o p e
w a s
u s e d
w i t h a
4 5 0
t o 4 9 0 - n m b a n d p a s s
f i l t e r ,
FT
5 1 0 b e a m s p l i t t e r , L P
5 2 0 b a r r i e r
f i l t e r ,
a n d a 1 0 0 - W
h a l og e n l a m p .
S e v e r a l
m o d i f i c a t i o n s
w e r e
m a d e i n t h e
a c r i d i n e
o r a n g e d i r e c t
c o u n t i n g p r o c e d u r e o f H o b b i e
e t
a l .
( 1 2 ) . N u c l e p o r e
p o l y c a r b o n a t e
m e m b r a n e s
( N u c l e p o r e
C o r p . ,
P l e a s a n -
t o n ,
C a l i f . ) o f 0 . 2 - , u m p o r e
s i z e w e r e
s t a i n e d f o r 2 0 m i n
i n
I r g a l a n
B l a c k ( C e i ba - G e i g y
C o r p . , G r e e n s b o r o ,
N . C . ) ,
a n d
t h e f i l t e r s , a f t e r
t h r e e r i n s e s i n d i s t i l l e d
w a t e r ,
w e r e u s e d
d a m p .
N o
s u r f a c t a n t
w a s
u s e d
9 .
A f t e r s t a i n i n g w i t h a c r i d i n e
o r a n g e ,
t h e f i l t e r
w a s
r i n s e d w i t h 0 . 8 5 s a l i n e . T h e
u n d e r - p a d
c e l l u l o s e
f i l t e r w a s n o t
e m p l o y e d .
T h e
d a m p
m e m b r a n e
f i l t e r
w a s p l a c e d o n
t h e s l i d e , a c o v e r s l i p
w a s
p l a c e d
d i r e c t l y
o n
t o p
o f
t h e
f i l t e r ,
a n d t h e
c e l l s
w e r e c o u n t e d
i m m e d i a t e l y .
A d r o p o f n o n f l u o r e s c e n t
o i l
o r s a l i n e
w a s a d d e d t o p r e v e n t r a p i d
d r y i n g o f
t h e m e m b r a n e .
S t a t i s t i c a l
a n a l y s i s . By u s i n g
a s t a n d a r d s t a t i s t i c a l
p r o g r a m ( S A S ) , a n a l y s i s
o f
v a r i a n c e
w a s
c o n d u c t e d .
C a l c u l a t e d f
v a l u e s w e r e
c o m p a r e d
w i t h
t a b u l a t e d
f
v a l u e s ( 1 7 ) .
D i f f e r e n c e s
i n
t h e d a t a w e r e
c o n s i d e r e d
s t a t i s t i c a l l y
s i g n i f i c a n t
a t
t h e 95 l e v e l
o f c o n f i d e n c e .
R E S U L T S
S u r v i v a l
o f
V .
c h o l e r a e
s e r o t y p e
0 1 . T h e e n v i -
r o n m e n t a l
V .
c h o l e r a e s e r o t y p e
0 1 s t r a i n
( W F /
1 )
s u r v i v e d
w e l l
i n
e s t u a r i n e
w a t e r s
a n d
s e d i -
m e n t s . F i g u r e 1 i l l u s t r a t e s t h e
e f f e c t o f t h e
i n o c u l u m
s i z e
o n
t h e
s u r v i v a l o f
t h e
o r g a n i s m i n
e s t u a r i n e
w a t e r s
a n d s e d i m e n t s . T h e i n i t i a l i n -
o c u l u m
h a d l i t t l e e f f e c t o n t h e
n u m b e r o f
o r g a n -
i s m s
r e c o v e r e d f r o m s t e r i l e s e d i m e n t s a f t e r 3
d a y s . A l t h o u g h
t h e n u m b e r o f v i a b l e
c e l l s r e c o v -
e r e d
w a s
i n i t i a l l y g r e a t e r i n
t h e e s t u a r i n e
w a t e r
a n d s e d i m e n t
c o n t a i n i n g
t h e
h i g h e s t
i n o c u l u m ,
t h e
d i f f e r e n c e s
w e r e
n o t
s i g n i f i c a n t .
A f t e r
7
d a y s , t h e
o r g a n i s m g r e w a b u n d a n t l y
i n
t h e
s t e r -
i l e
s e d i m e n t
a n d ( d a t a n o t
s h o w n )
c o u l d
b e
r e c o v e r e d
a f t e r
3 5 a n d 6 0
d a y s .
A t
3 5
d a y s ,
t h e
c o n c e n t r a t i o n o f
v i a b l e c e l l s h a d d e c l i n e d t o
1 0 4
c e l l s
p e r m l ,
a n d
b y
6 0
d a y s
t h e c o n c e n t r a t i o n
h a d f u r t h e r d e c r e a s e d
t o
1 0 2
c e l l s
p e r
m l . I n
n o n s t e r i l e
s e d i m e n t ,
v i a b l e c e l l s d e c l i n e d
r e l a -
t i v e l y q u i c k l y
a n d w e r e n o
l o n g e r
r e c o v e r a b l e
a f t e r
1 2
d a y s .
V i a b l e c e l l s a l s o d e c r e a s e d
i n
n o n s t e r i l e
w a t e r s , a l t h o u g h r e c o v e r y
wa s
p o s s i -
b l e f o r
u p
t o
3 5
d a y s .
T h e
s u r v i v a l
o f W F / 1 was e x a m i n e d
i n
s e d i -
m e n t s
( a n d w a t e r s )
c o l l e c t e d
f r o m
two
d i f f e r e n t
s i t e s
i n
A p a l a c h i c o l a B a y
d u r i n g J u l y .
S i t e 3 6
w a s t h e
l o c a t i o n
f r o m
w h i c h
t h e
o r g a n i s m
was
o r i g i n a l l y
i s o l a t e d ,
a n d
s i t e
2 9
was a n
a r e a
w i t h a
l o w e r
mean
s a l i n i t y
l o c a t e d
n e a r
t h e
m o u t h o f
V O L .
4 3 ,
1 9 8 2
onMay10,2013by
guest
http://aem.asm
.org/
Downlo
adedfrom
http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/ -
8/10/2019 Survival of Vibrio Cholerae And
4/8
A P P L . E N V I R O N .
M I C R O B I O L .
- - - - - - - - _ ,
1 0 5
K
n o n s t e r i l e
1 0
l o , L
1 0
0
1
2
3
4 5
6
7 8
9 1 0
1 1
1 2
Days
F I G .
2 .
S u r v i v a l
o f
V . c h o l e r a e 0 1
( W F / 1 )
i n s e d i m e n t s a n d
w a t e r s c o l l e c t e d
f r o m t w o
d i f f e r e n t s i t e s i n
A p a l a c h i c o l a
B a y a n d i n c u b a t e d
a t 2 0 C .
S i t e 2 9 h a d a
s a l i n i t y o f
6 / 0 0 ,
a n d s i t e 3 6 h a d a s a l i n i t y
o f 1 5 O I o .
T h e
v a l u e s a r e
e x p r e s s e d a s c o l o n y - f o r m i n g
u n i t s .
t h e
A p a l a c h i c o l a
R i v e r .
A l t h o u g h
t h e s a l i n i t i e s
o f
t h e
w a t e r s
w e r e
6
a n d
1 5 0 / 1 0 ,
t h e r e
w a s n o
s i g n i f i c a n t d i f f e r e n c e
i n t h e
s u r v i v a l
o f t h e
o r g a -
n i s m
i n
t h e
t w o
d i f f e r e n t
s e d i m e n t s
( F i g .
2 .
T h e s u r v i v a l
o f
W F / 1 v a r i e d
w i t h
t h e t i m e o f
y e a r
t h a t
w a t e r
a n d s e d i m e n t
w e r e c o l l e c t e d
( F i g . 3 .
A l t h o u g h t h e r e
w a s
n o s i g n i f i c a n t
d i f f e r -
e n c e i n
t h e s u r v i v a l o f
t h e
o r g a n i s m
i n
s t e r i l e
s e d i m e n t s
c o l l e c t e d
d u r i n g t h e
t h r e e m o n t h s ,
e a r l y r e c o v e r y ( l e s s
t h a n
5 d a y s ) o f v i a b l e c e l l s
i n
s t e r i l e w a t e r ,
n o n s t e r i l e
w a t e r , a n d
n o n s t e r i l e
s e d i m e n t
w a s g r e a t e s t
i n
t h e
S e p t e m b e r
s a m -
p l e s .
H o w e v e r ,
r e c o v e r y
i n n o n s t e r i l e w a t e r
a n d
n o n s t e r i l e s e d i m e n t
f r o m a n y s a m p l e
a f t e r
5
d a y s
w a s
n o t s i g n i f i c a n t l y
d i f f e r e n t .
T h e t e m p e r a t u r e a t
w h i c h t h e s u r v i v a l
c h a m -
b e r s
w e r e
i n c u b a t e d
h a d
a
s i g n i f i c a n t e f f e c t
o n
t h e
s u r v i v a l
o f
t h e o r g a n i s m
( F i g . 4 ) .
T h e o r g a -
n i s m
w a s o b s e r v e d
t o g r o w b e s t
a t 3 5 C i n s t e r i l e
e s t u a r i n e w a t e r
a n d
s e d i m e n t . H o w e v e r ,
i n
n o n -
s t e r i l e
s e d i m e n t a t t h i s t e m p e r a t u r e ,
t h e
o r g a -
n i s m
d i d n o t g r o w ,
n o r c o u l d
i t
b e
r e c o v e r e d
a f t e r
5
d a y s .
A t
i n c u b a t i o n t e m p e r a t u r e s
o f
2 5 C , r e c o v e r y
w a s
l i m i t e d
t o
5 d a y s i n
n o n s t e r -
i l e
s e d i m e n t a n d
1 0 d a y s i n
n o n s t e r i l e e s t u a r i n e
w a t e r .
A t
4 C , t h e
o r g a n i s m d i d
n o t g r o w
i n
s t e r i l e
s e d i m e n t s o r w a t e r s ,
n o r
w a s i t
r e c o v e r e d
a f t e r 5
d a y s ,
b u t
i t s u r v i v e d f o r
u p
t o
1 5 d a y s i n
n o n s t e r i l e
s e d i m e n t s
a n d
1 0
d a y s
i n
n o n s t e r i l e
e s t u a r i n e w a t e r .
S u r v i v a l
o f
o t h e r
V .
c h o l e r a e
s t r a i n s .
C o m p a r i -
s o n s o f
d i f f e r e n t V .
c h o l e r a e s t r a i n s r e v e a l e d n o
s i g n i f i c a n t
d i f f e r e n c e s
i n t h e
s u r v i v a l
o f s e r o t y p e
0 1
a n d
n o n - O 1 i s o l a t e s
i n
s t e r i l e
o r n o n s t e r i l e
s e d i m e n t s o r
w a t e r s .
N o
s i g n i f i c a n t
d i f f e r e n c e s
w e r e n o t e d i n
v i a b l e
c o u n t s
a m o n g
V . c h o l e r a e
0 1
( W F / 1 )
a n d
n o n - O 1
( W F / 2 3
o r W F / 5 4 )
s t r a i n s
i n
s e d i m e n t s
o r w a t e r s
( F i g .
5 ) .
T h e r e w a s
a l s o
n o s i g n i f i c a n t
d i f f e r e n c e
i n
t h e s u r v i v a l o f
W F / 1
a n d W F / 1 0 3
a s m e a s u r e d b y
v i a b l e
c o u n t s
( F i g .
6 .
S i m i l a r l y ,
t h e c l i n i c a l
V .
c h o l e r a e
0 1
i s o l a t e
s u r v i v e d
i n s e d i m e n t s
a n d
w a t e r
m u c h
l i k e
t h e
e n v i r o n m e n t a l
0 1 i s o l a t e
( F i g .
7 .
D i r e c t c o u n t s
o f 0 1 a n d n o n - O 1
s t r a i n s s h o w e d n o
s i g n i f i c a n t
d i f f e r e n c e s .
S t r a i n s W F / 1
a n d
W F / 1 0 8 ( F i g . 8 )
a n d W F / 1 a n d
W F / 1 1 1 ( F i g . 9 )
e x h i b i t e d t h e
s a m e r e s p o n s e
i n
s t e r i l e
e s t u a r i n e
w a t e r s .
S u r v i v a l o f
E .
c o l i .
I n
c o n t r a s t t o
t h e
V .
c h o l e r a e
s t r a i n s ,
E .
c o l i
A TCC 2 5 9 2 2
d i d
n o t
s u r v i v e
w e l l
i n
e s t u a r i n e
w a t e r s ( F i g . 6 ) a n d
c o u l d
n o t
b e
r e c o v e r e d
b y
p l a t i n g
o r
m o s t
p r o b a -
b l e
n u m b e r
p r o c e d u r e s
a f t e r 7
d a y s
i n
n o n s t e r i l e
w a t e r
a n d
1 4 d a y s
i n
s t e r i l e
w a t e r .
I n s t e r i l e
s e d i m e n t ,
E .
c o l i
d i d n o t
s u r v i v e
a s
w e l l a s V .
c h o l e r a e ;
b u t
i n
n o n s t e r i l e
s e d i m e n t ,
i t a c t u a l l y
s u r v i v e d b e t t e r
t h a n
t h e V . c h o l e r a e
s t r a i n s .
When a
f r e s h l y
i s o l a t e d
human s t r a i n
o f
E .
c o l i
w a s
e x a m i n e d ,
i t a l s o d i d n o t
s u r v i v e
a s
w e l l
a s t h e
V .
c h o l e r a e s t r a i n s
i n s t e r i l e e s t u a r i n e
w a t e r
( F i g .
9 ) .
A l t h o u g h
E .
c o l i
m o s t p r o b a b l e
n u m b e r s
w e r e
s t a t i s t i c a l l y
t h e
s a m e
a s
t h e
v i a b l e
c o u n t s ,
t h e
d i r e c t c o u n t s
w e r e much
h i g h e r .
A l l
a t t e m p t s
t o
r e s u s c i t a t e
t h e
i n t a c t
E .
c o l i
c e l l s
b y
u s i n g
l a c t o s e
e n r i c h m e n t
f a i l e d .
D i r e c t
a n d
v i a b l e c o u n t s .
T h e
d i f f e r e n c e s
b e -
t w e e n d i r e c t
c o u n t s
a n d v i a b l e
c o u n t s o f
V .
c h o l e r a e
s t r a i n s
i n
s t e r i l e
e s t u a r i n e
w a t e r w e r e
f a r
l e s s
t h a n
w i t h E .
c o l i
s t r a i n s .
V i a b l e
c o u n t s
5 8 0
HOOD
AND
N E S S
onMay10,2013by
guest
http://aem.asm
.org/
Downlo
adedfrom
http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/http://aem.asm.org/ -
8/10/2019 Survival of Vibrio Cholerae And
5/8
V .
CH O L E R A E
AND
E .
COLI IN ES TUARIES
10 4
1 0
1 0 2
0 1
0
4 * C
1 0
1 0
1 0
1 0 3
1 0 2
1 0
1
2
3
4
5 6
7
8
9 1 0 n 1 1 2 1 3
1
1 5
D a y s
F I G . 4 .
E f f e c t
o f
t e m p e r a t u r e
o n
t h e
s u r v i v a l
o f V .
c h o l e r a e
0 1
( W F / 1 )
i n
e s t u a r i n e
w a t e r s a n d
s e d i m e n t s .
T h e
v a l u e s a r e
e x p r e s s e d
a s
c o l o n y - f o r m i n g
u n i t s .
S y m b o l s :
* ,
s t e r i l e
s e d i m e n t ;
0 ,
s t e r i l e
e s t u a r i n e
w a t e r ;
O ,
n o n s t e r i l e
s e d i m e n t ;
0 ,
n o n s t e r i l e
e s t u a r i n e
w a t e r .
1
2
3 4
5
6
7
8
9 1 n 12
Days
F I G .
3 .
S u r v i v a l
o f
V .
c h o l e r a e
01
( W F / 1 )
i n
e s t u a -
r i n e w a t e r s a n d
s e d i m e n t s c o l l e c t e d
d u r i n g
3 d i f f e r e n t
m o n t h s
a n d
i n c u b a t e d a t
2 0 C . T he v a l u e s
are
ex -
p r e s s e d as
c o l o n y - f o r m i n g
u n i t s .
S y m b o l s :
* ,
s t e r i l e
s e d i m e n t ; 0
s t e r i l e e s t u a r i n e
w a t e r ;
0
n o n s t e r i l e
s e d i m e n t ;
0
n o n s t e r i l e
e s t u a r i n e
w a t e r .
o f
V .
c h o l e r a e
s t r a i n s
were
1
t o
0 . 5
l o g
l e s s
t h a n
d i r e c t
c o u n t s , a n d
t h e
d i f f e r e n c e
r e m a i n e d
con-
s t a n t
over
t h e
e x p e r i m e n t a l
p e r i o d
( F i g .
8
a n d
9 ) .
H o w e v e r ,
v i a b l e
c o u n t s
o f
E .
c o l i
were
a s
muc h
as
3
l o g s
l o w e r
t h a n
d i r e c t
c o u n t s ; o v e r
t h e
e x p e r i m e n t a l
p e r i o d , b o t h
d e c l i n e d
a t
d i f f e r e n t
r a t e s
( F i g .
9 ) .
T h e
g r e a t e s t
d i f f e r e n c e
b e t w e e n
v i a b l e
a n d
d i r e c t
c o u n t s
o f
V .
c h o l e r a e
was
o b s e r v e d
w i t h
t h e
a d a p t e d 01
s t r a i n
( F i g . 9 ) .
V i a b l e
c o u n t s
o f
t h i s
o r g a n i s m
i n
s t e r i l e
e s t u a r i n e
water
were 2
l o g s
l o w e r t h a n
d i r e c t
c o u n t s
a t
d a y
0 .
H o w e v e r ,
a f t e r 2
d a y s ,
t h e
d i f f e r e n c e i n
d i r e c t
a n d
v i a b l e
c o u n t s
was
o n l y 0 . 5
l o g . W h y
t h i s
response
occurs i s
u n c e r t a i n , b u t
t h e
i n i t i a l
l o s s o f
v i a b i l i -
t y
o f
s u c h
a
l a r g e p o p u l a t i o n
o f c e l l s
may b e
a
r e f l e c t i o n
o f
s t r e s s
f a c t o r s .
T h e s e
a d a p t e d
01
c e l l s
were
n u t r i e n t
s t a r v e d , r e v i t a l i z e d
on
T r y p -
t i c a s e
soy
b r o t h ,
p l a c e d i n
e s t u a r i n e
w a t e r ,
a n d
t h e n
p l a t e d
o n t o
b r a i n
h e a r t
i n f u s i o n
agar.
E x p o -
s u r e t o
s u c h
e x t r e m e
c o n d i t i o n s
may r e s u l t
i n
a n
i n i t i a l
l o s s
o f
v i a b i l i t y
on
b r a i n
h e a r t
i n f u s i o n
a g a r .
DISCUSSION
S t r a i n s o f
V .
c h o l e r a e ,
i n c l u d i n g
e n v i r o n m e n -
t a l ,
c l i n i c a l ,
a n d
s e r o t y p e
01
a n d
n o n - O 1
s t r a i n s , s h o w e d
n o d i f f e r e n c e s
i n s u r v i v a l
r a t e s
i n
e s t u a r i n e
w a t e r s
a n d
s e d i m e n t s .
T h e
o b s e r v e d
g r o w t h
o f V .
c h o l e r a e
i n
s t e r i l e
s e d i m e n t
wa s
p r o b a b l y
d u e
t o
t h e
a b s e n c e
o f
c o m p e t i n g
o r g a n -
i s m s
a n d
t h e
a b u n d a n c e
o f
a v a i l a b l e
n u t r i e n t s .
G e r b a
a n d
McLeod
( 1 0 ) h a v e d e m o n s t r a t e d
t h a t
a u t o c l a v i n g
r e s u l t s i n
t h e
r e l e a s e
o f
b o u n d
n u t r i -
e n t s
f r o m s e d i m e n t s .
T h e s e n u t r i e n t s
w o u l d
n o t
b e
a v a i l a b l e
i n t h e
n a t u r a l e n v i r o n m e n t .
T h e r e -
f o r e ,
t h e
o b s e r v e d
g r o w t h
o f V . c h o l e r a e
i n
s t e r i l e
s e d i m e n t
w o u l d b e
d u e , i n
p a r t ,
t o
t h e
a v a i l a b i l i t y
o f s e d i m e n t - b o u n d
n u t r i e n t s .
I
I
z
V O L .
4 3 ,
1 9 8 2
5 8 1
onMay10,2013by
guest
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.org/
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8/10/2019 Survival of Vibrio Cholerae And
6/8
s t e r i l e
s e d i m e n t
E
1 0 2
0
n o n s t e r i l e
water
o 3
1 0
2
10 1
1
2
3
4 5 6
7
8
9
1 0 1 1
1 2
D a y s
F I G . 5 . S u r v i v a l
o f V .
c h o l e r a e
0 1
( W F / 1 )
a n d t w o
n o n - 0 1
s t r a i n s
( W F / 2 3
a n d
W F / 5 4 )
i n
e s t u a r i n e w a t e r s
a n d
s e d i m e n t s a t
2 0 C .
T h e
v a l u e s a r e
e x p r e s s e d
a s
c o l o n y - f o r m i n g
u n i t s .
S y m b o l s :
O , 0 1 ;
* ,
n o n - 0 1
( W F / 2 3 ) ;
* ,
n o n - 0 1
( W F / 5 4 ) .
G r o w t h
a n d
l o n g
s u r v i v a l were
a l s o n o t e d i n
s t e r i l e
e s t u a r i n e
w a t e r s
a n d a r e
p r o b a b l y
d u e
t o
t h e a b s e n c e
o f
c o m p e t i n g
o r g a n i s m s .
I n n o n s t e r -
i l e
w a t e r s ,
t h e
o r g a n i s m s e x h i b i t e d
g r o w t h
a n d
s u r v i v e d
f o r
l o n g p e r i o d s
o f
t i m e ,
b u t
t h e y
d i d
n o t
s u r v i v e
e s p e c i a l l y
w e l l i n
s e d i m e n t s .
T h e s e
r e s u l t s
s u g g e s t
t h a t
s e r o t y p e
01
a n d
n o n - O 1
o r g a n i s m s
a re b o t h
w e l l
a d a p t e d
t o a n d
c a p a b l e
o f
l i v i n g
i n
e s t u a r i n e
w a t e r s .
F u r t h e r m o r e ,
t h e
c l i n i c a l
i s o l a t e , e x h i b i t i n g
t h e same s u r v i v a l
r e -
sponse,
a l s o
appears
t o
b e
w e l l
a d a p t e d
t o
e s t u a r i n e
w a t e r s .
I n
c o n t r a s t t o
V .
c h o l e r a e ,
s t r a i n s
o f
E .
c o l i
d i d
n o t
s u r v i v e w e l l i n
e s t u a r i n e
w a t e r s .
T h e f a c t
t h a t E .
c o l i
d o e s n o t s u r v i v e
w e l l i n
seawater
a l o n e
b u t
i s
a b l e t o
s u r v i v e
b e t t e r
i n
t h e
pres en ce
o f m a r i n e
s e d i m e n t s
h a s
b e e n
p r e v i o u s l y
d e m o n -
A P P L . E N V I R O N .
M I C R O B I O L .
1 0
e
10 4
1 0 3
2
10
l o
1 0 3
0 2
10
0
10 7
10 5
O
4
0
1 2
3
4 5 6 7
8 9 1 0
1 1 1 2
1 3
1 4
D a y s
F I G . 6 . S u r v i v a l o f V . c h o l e r a e
01
( W F / 1 ) ,
n o n - 0 1
( W F / 1 0 3 ) ,
a n d
E .
c o l i
A T CC 2 5 9 2 2
i n
e s t u a r i n e
w a t e r s
a n d
s e d i m e n t s a t
2 0 C .
T h e v a l u e s are
e x p r e s s e d
a s
c o l o n y - f o r m i n g
u n i t s .
s t r a t e d
( 1 0 ) .
Our
r e s u l t s
c o n f i r m t h a t
a
s i m i l a r
r es p on s e o cc u rs
i n
e s t u a r i n e
w a t e r s
a n d
s e d i -
m e n t s .
E .
c o l i
was
r e c o v e r a b l e
f r o m
e s t u a r i n e
s e d i m e n t s
f o r
c o n s i d e r a b l y l o n g e r
t h a n
f r o m
w a t e r s .
F u r t h e r m o r e ,
i t s u r v i v e d
e v e n b e t t e r
i n
n o n s t e r i l e
s e d i m e n t s
t h a n V .
c h o l e r a e .
T h e
f i n d i n g
t h a t E .
c o l i d o e s n o t s u r v i v e as
w e l l
as V .
c h o l e r a e
i n
e s t u a r i n e
w a t e r s h a s
s i g n i f i c a n t
i m p l i c a t i o n s
i n t e r m s
o f w a t e r
a n d
s h e l l f i s h
s a f e t y .
S i n c e
f e c a l
c o l i f o r m
E .
c o l i
l e v e l s are u s e d
t o
i n d i c a t e t h e
q u a l i t y
o f
w a t e r
a n d
s h e l l f i s h ,
t h e
q u e s t i o n
a r i s e s a s
t o t h e v a l i d -
i t y
o f
f e c a l
c o l i f o r m s a s
i n d i c a t o r s
o f
V .
c h o -
l e r a e .
One o f
t h e
b a s i c c r i t e r i a
f o r
a
g o o d
i n d i c a -
t o r
o r g a n i s m
i s t h a t
t h e i n d i c a t o r m u st
s u r v i v e a s
l o n g
as t h e
p a t h o g e n .
T h e
r e s u l t s o f
t h i s
s t u d y
d e m o n s t r a t e
t h a t
E .
c o l i
d o e s
n o t s u r v i v e i n
5 8 2
HOOD AND
N E S S
onMay10,2013by
guest
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.org/
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8/10/2019 Survival of Vibrio Cholerae And
7/8
V . C H O L E R A E AND
E .
COLI IN ES TUARIES
5 8 3
E .
c o l i
Human
Isoate
E
10 '
1 0 8
0 1
( C l i n i l
I s o l a t e
10
7
1 0
10
1 0 5
10 4
10 3
1
2
3
4
5
6
7
9
1 0
1 2
1 3
14
1 5
Days
F I G .
7 . S u r v i v a l
o f
V . c h o l e r a e
0 1
( W F / 1 )
a n d a
c l i n i c a l V . c h o l e r a e
0 1
s t r a i n
i n
e s t u a r i n e
w a t e r s a n d
s e d i m e n t s a t 2 0 0 C .
T h e v a l u e s are
e x p r e s s e d
a s
c o l o -
n y - f o r m i n g
u n i t s .
S y m b o l s :
* ,
s t e r i l e
s e d i m e n t ; * ,
s t e r i l e
e s t u a r i n e
w a t e r ;
O ,
n o n s t e r i l e
s e d i m e n t ;
0
n o n s t e r i l e e s t u a r i n e
w a t e r .
e s t u a r i n e
w a t e r
as w e l l
as
V .
c h o l e r a e .
T h i s
s u g g e s t s
t h a t
m e a s u r i n g
t h e l e v e l s
o f f e c a l c o l i -
f o r m s i n
t h e
w a t e r c o l u m n may n o t
b e a d e q u a t e
f o r m o n i t o r i n g
t h e
presence
o f
V .
c h o l e r a e .
F u r t h e r m o r e , t h e r e i s i n c r e a s i n g
e v i d e n c e
f r o m
e n v i r o n m e n t a l
s t u d i e s
t h a t
f e c a l c o l i f o r m l e v e l s
3
0
1 0 2 .
1 0 7 .
1 0
1 o 5
1 0 o
1 0 2
z
1 0 7
1 o 6
1 o 5
1 0 4
1 0 2
1 0 1
1 0 7
1 0 6
1 0 5
1 0
1 0 1 .
1 0 6
E
z
0
ODC
. . . . . . . . . . . . . .
o
N o n - 0 1
IC
o
N
o n - 0
1 0 4
A O D C
a t p N v
NON
01
ODC
F U
Adapted
01
ODC
CFU-
0 1
ODc
CF U
2
4
6
DAYS
a 10
12 14
F I G .
9 .
S u r v i v a l
o f
V .
c h o l e r a e
01 ( W F / 1 ) ,
a n
e s t u a r y - a d a p t e d
s t r a i n
( W F / 1 ) ,
a n o n - O 1
s t r a i n
( W F /
1 1 1 ) , a n d a n
E . c o l i human
i s o l a t e i n s t e r i l e e s t u a r i n e
w a t e r a t
2 0 C . T h e v a l u e s are
e x p r e s s e d as
a c r i d i n e
orange
d i r e c t
c o u n t s
(AODC)
a n d
c o l o n y - f o r m i n g
u n i t s
( C F U ) .
1
2
3
4 5
6
7
Day s
F I G .
8 .
S u r v i v a l o f
V .
c h o l e r a e
0 1 ( W F / 1 )
a n d a
n o n - O 1
s t r a i n
( W F / 1 0 8 )
i n
e s t u a r i n e
w a t e r s
a t
2 0 C .
T h e v a l u e s are
e x p r e s s e d
a s
a c r i d i n e
orange
d i r e c t
c o u n t s
(AODC)
a n d
c o l o n y - f o r m i n g
u n i t s
( C F U ) .
S y m b o l s :
U
s t e r i l e s e d i m e n t ; 0
s t e r i l e
e s t u a r i n e
w a t e r ;
l,
n o n s t e r i l e
s e d i m e n t ;
0
n o n s t e r i l e e s t u a r i n e
w a t e r .
i n
e s t u a r i n e w a t e r s
d o
n o t
c o r r e l a t e
w e l l
w i t h V .
c h o l e r a e l e v e l s
( 8 ,
1 3 ,
1 4 ) .
T h u s ,
i f
f e c a l c o l i f o r m
l e v e l s
i n
e s t u a r i n e
w a t e r s
d o n o t
i n d i c a t e
t h e
presence
o f
V .
c h o l e r a e ,
o t h e r
means o f
d e t e r -
m i n i n g p o t e n t i a l l y
d a n g e r o u s
l e v e l s
o f
V . c h o -
l e r a e
i n
e s t u a r i n e
w a t e r s
a n d
s h e l l f i s h
mus t
b e
f o u n d .
A C K N O W L E D G M E N T S
W e
g r a t e f u l l y
a c k n o w l e d g e
R on
B a k e r ,
J o h n
C h e n e y ,
a n d
K a t h y
W i l l i a m s
f o r t h e i r
e x c e l l e n t
t e c h n i c a l
s u p p o r t ,
t h e
V O L . 4 3 ,
1 9 8 2
onMay10,2013by
guest
http://aem.asm
.org/
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adedfrom
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8/10/2019 Survival of Vibrio Cholerae And
8/8
5 8 4
HOOD
AND N ESS
F l o r i d a S t a t e
D e p a r t m e n t o f N a t u r al R e s o u r c e s
f o r t h e i r
l o g i s t i c a l s u p p o r t ,
a n d
R . R .
C o l w e l l
f o r
h e l p f u l
d i s c u s s i o n s .
T h e
w o r k
i s
a
r e s u l t
o f
r e s e a r c h s p o n s o r e d b y
F l o r i d a
S e a
G r a n t ,
N a t i o n a l
O c e a n i c
a n d
A t m o s p h e r i c A d m i n i s t r a t i o n ,
O f f i c e
o f S e a G r a n t , D e p a r t m e n t
o f C o m m e r c e , u n d e r g r a n t
SG NA
8 0 A A - D - 0 0 0 3 8 .
L I T E R A T U R E
CITED
1 . A s s o c i a t i o n
o f A n a l y t i c a l C h e m i s t s .
1 9 7 5 .
O f f i c i a l m e t h o d s
o f a n a l y s i s o f t h e
A s s o c i a t i o n o f A n a l y t i c a l
C h e m i s t s , 1 2 t h
e d .
A s s o c i a t i o n o f A n a l y t i c a l
C h e m i s t s , W a s h i n g t o n ,
D . C .
2 . B l a k e ,
P . A . , D . T . A l H e g r a ,
J .
D .
S n y d e r ,
T .
J . B a r r e t t ,
L .
M c F a r l a n d , C .
T .
C a r a w a y ,
J .
C . F e e l e y ,
J .
P .
C r a i g ,
J .
V . L e e ,
N . D .
P u h r ,
a n d
R . A .
F e l d m a n .
1 9 8 0 . C h o l -
e r a - a
p o s s i b l e
e n d e m i c
f o c u s
i n t h e U n i t e d S t a t e s .
N .
E n g l .
J . M e d .
3 0 2 : 3 0 5 - 3 0 9 .
3 . C a r l u c c i ,
A .
F . ,
a n d
D . P r a m e r . 1 9 5 9 . F a c t o r s
a f f e c t i n g
s u r v i v a l o f
b a c t e r i a
i n
s e a w a t e r .
A p p l .
M i c r o b i o l . 7 : 3 8 8 -
3 9 2 .
4 .
C e n t e r s
f o r
D i s e a s e
C o n t r o l .
1 9 7 8 .
V i b r i o
c h o l e r a e - L o u i -
s i a n a .
M o r b i d .
M o r t a l .
W e e k l y R e p .
2 7 : 3 6 7 .
5 .
C e n t e r s
f o r D i s e a s e
C o n t r o l .
1 9 7 9 .
N o n - O 1
V i b r i o
c h o l e r a e
i n f e c t i o n s - F l o r i d a .
M o r b i d .
M o r t a l .
W e e k l y
R e p .
2 8 : 5 7 1 - 5 7 2 .
6 . C e n t e r s
f o r
D i s e a s e
C o n t r o l .
1 9 8 0 . C h o l e r a e - F l o r i d a .
M o r b i d . M o r t a l .
W e e k l y R e p .
2 9 : 6 0 1 .
7 .
C o l w e l l ,
R .
R . ,
J . K a p e r ,
a n d
S .
W.
J o s e p h .
1 9 7 7 . V i b r i o
c h o l e r a e ,
V i b r i o
p a r a h a e m o l y t i c u s ,
a n d
o t h e r v i b r i o s :
o c c u r r e n c e
a n d d i s t r i b u t i o n
i n
C h e s a p e a k e
B a y .
S c i e n c e
1 9 8 : 3 9 4 - 3 9 6 .
8 .
C o l w e l l ,
R . R . , R . J . S e i d l e r , J . K a p e r ,
S .
W.
J o s e p h ,
S .
G a r g e s ,
H .
L o c k m a n ,
D .
M a n e v a l ,
H .
B r a d f o r d ,
N .
R o b -
e r t s ,
E .
R e m m e r s ,
I .
H u q ,
a n d
A .
H u q .
1 9 8 1 . O c c u r r e n c e
o f V i b r i o
c h o l e r a e s e r o t y p e
O 1
i n
M a r y l a n d
a n d
L o u i s i a n a
e s t u a r i e s .
A p p l .
E n v i r o n . M i c r o b i o l .
4 1 : 5 5 5 - 5 5 8 .
9 .
D a l e y ,
R .
J .
1 9 7 9 . D i r e c t
e p i f l u o r e s c e n c e
e n u m e r a t i o n
o f
n a t u r a l
a q u a t i c
b a c t e r i a : u s e s , l i m i t a t i o n s ,
a n d
c o m p a r a -
t i v e a c c u r a c y , p . 2 9 - 4 5 .
I n
J .
C o s t e r t o n
a n d
R .
R .
C o l w e l l
( e d . ) ,
N a t i v e a q u a t i c
b a c t e r i a : e n u m e r a t i o n , a c t i v i t y ,
a n d
e c o l o g y .
A m e r i c a n S o c i e t y f o r T e s t i n g
a n d
M a t e r i a l s ,
W a s h i n g t o n ,
D . C .
1 0 .
G e r b a ,
C . P . ,
a n d
J . S .
M c L e o d . 1 9 7 6 . E f f e c t s o f s e d i -
m e n t s o n s u r v i v a l
o f
E s c h e r i c h i a
c o l i i n m a r i n e
w a t e r s .
A p p i .
E n v i r o n .
M i c r o b i o l .
3 2 : 1 1 4 - 1 2 0 .
1 1 . G r e e n be r g, A .
E .
1 9 5 6 .
S u r v i v a l o f
e n t e r i c
o r g a n i s m s
i n
s e a w a t e r .
P u b l i c H e a l t h
R e p .
7 1 : 7 7 - 8 6 .
1 2 .
H o b b i e ,
J . E . ,
R . J .
D a l e y ,
a n d S . J a s p e r . 1 9 7 7 . U s e
o f
n u c l e p o r e f i l t e r s
f o r c o u n t i n g
b a c t e r i a b y
f l u o r e s c e n c e
m i c r o s c o p y .
A p p l .
E n v i r o n .
M i c r o b i o l .
3 3 : 1 2 2 5 - 1 2 2 8 .
1 3 . H o o d ,
M .
A . , G . E . N e s s ,
a n d G . E .
R o d r i c k .
1 9 8 1 . I s o l a -
t i o n
o f V i b r i o
c h o l e r a e s e r o t y p e 0 1 f r o m
t h e e a s t e r n
o y s t e r ,
C r a s s o s t r e a
v i r g i n i c a .
A p p l .
E n v i r o n .
M i c r o b i o l .
4 1 : 5 5 9 - 5 6 0 .
1 4 .
K a p e r , J . , H . L o c k m a n , R .
R . C o l w e l l , a n d
S .
W. J o s e p h .
1 9 7 9 . E c o l o g y ,
s e r o l o g y ,
a n d e n t e r o t o x i n
p r o d u c t i o n
o f
V i b r i o c h o l e r a e i n C h e s a p e a k e B a y .
A p p l . E n v i r o n . M i -
c r o b i o l .
3 7 : 9 1 - 1 0 3 .
1 5 . M i t c h e l l , R . 1 9 6 8 . F a c t o r s
a f f e c t i n g t h e
d e c l i n e
o f n on -
m a r i n e m i c r o o r g a n i s m s
i n
s e a w a t e r .
W a t e r
R e s . 2 : 5 3 5 -
5 4 3 .
1 6 .
P o l l i t z e r ,
R .
1 9 5 9 . C h o l e r a ,
p .
1 8 5 - 1 8 6 .
I n W o r l d H e a l t h
O r g a n i z a t i o n
m o n o g r a p h s e r i e s
n o .
4 3 .
W o r l d H e a l t h
O r g a n i z a t i o n ,
G e n e v a .
1 7 .
S n e d e c o r ,
G . W . , a n d
W.
G .
C o c h r a n .
1 9 6 7 .
S t a t i s t i c a l
m e t h o d s ,
6 t h e d . I o w a S t a t e U n i v e r s i t y
P r e s s , A m e s .
1 8 .
T w e d t ,
R . M . , J . M.
M a d d e n , J . M . H u n t , D .
W .
F r a n c i s ,
J .
T .
P e e l e r , A .
P .
D u r a n ,
W. 0 .
H e b e r t ,
S . G . M c C a y ,
C .
N . R o d e r i c k ,
G .
T .
S p i t e ,
a n d
T .
J . W a z e n s k i .
1 9 8 1 .
C h a r a c t e r i z a t i o n
o f V i b r i o c h o l e r a e i s o l a t e d
f r o m o y s t e r s .
A p p l .
E n v i r o n . M i c r o b i o l .
4 1 : 1 4 7 5 - 1 4 7 8 .
1 9 .
W e i s s , C .
M .
1 9 5 1 . A d s o r p t i o n o f E . c o l i
o n
r i v e r a n d
e s t u a r i n e
s i l t s .
S e w a g e
I n d .
W a s t e s .
2 3 : 2 2 7 - 2 3 7 .
2 0 .
W e i s s m a n ,
J . B . , W. E . D e W i t t ,
J . T h o m p s o n , C . N .
M u c h n i c k ,
B .
L .
P o r t n o y , J . C .
F e e l e y , a n d
E .
J . G a n g a r o -
s a .
1 9 7 5 .
A
c a s e o f c h o l e r a
i n
T e x a s ,
1 9 7 3 .
Am.
J .
E p i d e m i o l .
1 0 0 : 4 8 7 - 4 9 8 .
A P P L . E N V I R O N . M I C R O B I O L .
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