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s
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m
e
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ls
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c
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lt
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les
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m
e
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a
n
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a
u
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d
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c
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re
a
d
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b
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th
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a
n
d
s.
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e
ID
b
ro
t
h
sta
b
il
ize
s
th
e
b
a
c
teria
'
s
c
o
n
d
i
ti
o
n
b
e
f
o
re
b
e
in
g
g
ro
w
n
o
n
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u
tri
e
n
t
a
g
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r
m
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ia
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K
ey
w
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d
s
:
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b
r
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cCo
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k
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m
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lmo
n
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typ
h
i
SDS
-
PAGE
T
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is i
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a
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a
c
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a
rticle
u
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d
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CC B
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C
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A
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Su
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Dep
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Me
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Hea
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s
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Salm
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ac
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o
n
m
ed
ia
ap
p
ea
r
s
co
n
v
e
x
,
s
lip
p
er
y
,
an
d
clea
r
[
5
]
,
[
6
]
.
OM
P
an
tig
en
s
ar
e
lo
ca
ted
o
n
th
e
s
u
r
f
ac
e
o
f
g
r
am
-
n
eg
ativ
e
b
ac
ter
ia.
T
h
ey
a
r
e
k
n
o
wn
to
b
e
g
o
o
d
at
in
d
u
cin
g
im
m
u
n
ity
ag
ai
n
s
t
S
a
lmo
n
ella
typ
h
i
b
y
ac
tiv
atin
g
th
e
ce
llu
lar
im
m
u
n
e
s
y
s
tem
.
R
esear
ch
er
s
ar
e
s
tu
d
y
in
g
OM
P
as
a
p
o
ten
tial
v
ac
cin
e
ca
n
d
id
ate
ag
ain
s
t
ty
p
h
o
id
f
e
v
er
.
T
h
e
p
ath
o
g
en
icit
y
o
f
S
a
lmo
n
ella
in
in
d
u
cin
g
d
iar
r
h
ea
is
a
co
m
p
le
x
p
h
en
o
m
en
o
n
th
at
in
v
o
lv
es
s
ev
er
al
m
ec
h
an
is
m
s
,
in
clu
d
in
g
th
e
p
r
o
d
u
ctio
n
o
f
en
ter
o
to
x
in
[
7
]
.
Stu
d
ies
h
av
e
s
h
o
wn
th
at
v
ac
cin
es
b
ased
o
n
OM
P
ar
e
e
f
f
ec
tiv
e
i
n
in
d
u
cin
g
p
r
o
tectiv
e
an
tib
o
d
ies
ag
ain
s
t
ty
p
h
o
id
,
m
ak
in
g
OM
P
f
r
o
m
S
a
lmo
n
ella
a
p
o
ten
tial
ca
n
d
id
ate
f
o
r
p
r
o
tectio
n
.
Ho
wev
er
,
v
ac
cin
es
b
ased
o
n
p
o
ly
s
ac
ch
a
r
id
es
an
d
O
a
n
tig
en
s
m
a
y
n
o
t
b
e
ef
f
ec
ti
v
e
an
d
ca
n
e
v
en
in
cr
ea
s
e
s
u
s
ce
p
tib
ilit
y
to
ex
tr
ac
ellu
lar
S
a
lmo
n
ella
g
r
o
wth
,
wh
ich
ca
n
b
e
d
an
g
e
r
o
u
s
.
I
n
r
ec
en
t
y
ea
r
s
,
r
esear
ch
e
r
s
h
av
e
co
n
d
u
cted
ex
ten
s
iv
e
s
tu
d
ies
o
n
th
e
m
o
lecu
lar
s
tr
u
ctu
r
e
an
d
f
u
n
ctio
n
o
f
OM
P
f
r
o
m
S
a
lmo
n
ella
as
a
p
o
ten
tial
v
ac
cin
e
ca
n
d
id
ate,
v
ir
u
len
ce
f
ac
to
r
,
a
n
d
d
ia
g
n
o
s
tic
an
tig
e
n
.
Desp
it
e
th
is
,
o
n
ly
a
lim
ited
n
u
m
b
er
o
f
OM
Ps
h
av
e
b
ee
n
f
u
lly
ch
ar
ac
ter
ize
d
s
o
f
ar
[
8
]
.
SDS
-
PAGE
,
wh
ich
s
tan
d
s
f
o
r
s
o
d
iu
m
d
o
d
ec
y
l
s
u
lf
ate
-
p
o
ly
ac
r
y
lam
id
e
g
el
elec
tr
o
p
h
o
r
esis
,
is
a
tech
n
iq
u
e
u
s
ed
to
s
ep
ar
ate
p
r
o
tein
s
b
ased
o
n
th
ei
r
m
o
lec
u
lar
weig
h
t.
T
h
is
m
eth
o
d
u
ti
lizes
th
e
ab
ilit
y
o
f
p
r
o
tein
s
to
m
ig
r
ate
to
war
d
s
an
elec
tr
o
d
e
in
th
e
p
o
l
y
ac
r
y
l
am
id
e
g
el
b
ased
o
n
th
eir
s
iz
e
u
n
d
e
r
an
elec
tr
ic
cu
r
r
en
t
[
9
]
,
[
1
0
]
.
Pro
tein
s
a
r
e
s
ep
ar
ated
b
y
p
ass
in
g
th
r
o
u
g
h
a
g
el
m
atr
i
x
m
ad
e
o
f
ac
r
y
lam
id
e
d
u
r
in
g
elec
tr
o
p
h
o
r
esis
.
T
h
is
s
ep
ar
atio
n
r
elies
o
n
th
e
m
o
lecu
lar
m
as
s
o
f
th
e
p
r
o
tein
s
,
with
s
m
aller
m
o
lecu
les
m
o
v
in
g
t
h
r
o
u
g
h
t
h
e
g
e
l
f
a
s
t
e
r
t
h
a
n
l
a
r
g
e
r
m
o
l
e
c
u
l
e
s
.
E
l
e
c
t
r
o
p
h
o
r
e
s
i
s
i
s
a
h
i
g
h
l
y
s
e
n
s
i
t
i
v
e
t
e
c
h
n
i
q
u
e
t
h
a
t
d
o
e
s
n
o
t
a
l
t
e
r
t
h
e
s
t
r
u
c
t
u
r
e
o
f
p
r
o
t
e
i
n
s
,
e
v
e
n
w
i
t
h
s
l
i
g
h
t
v
a
r
i
a
n
c
e
s
i
n
c
h
a
r
g
e
a
n
d
m
o
l
e
c
u
l
a
r
w
e
i
g
h
t
a
f
f
e
c
t
i
n
g
t
h
e
i
r
m
o
v
e
m
e
n
t
[
1
1
]
.
Gr
o
wth
m
ed
ia
is
a
s
u
b
s
tan
ce
th
at
co
n
tain
s
n
u
tr
ien
ts
ess
en
tial f
o
r
th
e
g
r
o
wth
o
f
m
icr
o
o
r
g
an
i
s
m
s
.
T
h
i
s
m
ed
iu
m
ca
n
b
e
u
s
ed
f
o
r
is
o
latin
g
,
id
en
tif
y
in
g
,
a
n
d
cr
e
atin
g
p
u
r
e
cu
ltu
r
es
o
f
m
icr
o
o
r
g
a
n
is
m
s
,
wh
o
s
e
n
u
tr
itio
n
al
r
eq
u
ir
em
en
ts
v
ar
y
d
ep
en
d
i
n
g
o
n
th
e
ty
p
e.
Gen
er
ally
,
th
ey
r
eq
u
ir
e
m
ac
r
o
n
u
tr
ie
n
ts
,
m
icr
o
n
u
tr
ien
ts
,
v
itam
in
s
,
an
d
m
in
er
als
to
th
r
iv
e.
Ma
cr
o
n
u
tr
ie
n
ts
s
u
ch
as
ca
r
b
o
n
,
h
y
d
r
o
g
e
n
,
an
d
n
itr
o
g
e
n
ar
e
n
ec
ess
ar
y
in
lar
g
er
q
u
an
titi
es,
wh
ile
m
icr
o
n
u
tr
ien
ts
ar
e
r
eq
u
ir
ed
in
s
m
aller
am
o
u
n
ts
an
d
c
o
n
tr
ib
u
te
to
e
n
zy
m
e
f
u
n
ctio
n
a
n
d
p
r
o
tein
s
tr
u
ctu
r
e.
Mic
r
o
n
u
tr
ie
n
ts
ar
e
ca
teg
o
r
ize
d
in
to
t
r
ac
e
elem
en
ts
an
d
g
r
o
wth
f
ac
t
o
r
s
,
with
tr
ac
e
elem
en
ts
s
er
v
in
g
as
en
zy
m
e
co
f
ac
to
r
s
,
in
clu
d
in
g
ir
o
n
,
m
an
g
a
n
ese,
an
d
zin
c
.
C
u
ltu
r
e
m
ed
ia
is
a
m
at
er
ial
co
n
s
is
tin
g
o
f
a
m
ix
tu
r
e
o
f
n
u
tr
ien
ts
u
s
ed
f
o
r
cu
ltu
r
es
o
f
a
m
icr
o
o
r
g
an
is
m
.
T
h
e
cr
ea
tio
n
o
f
cu
ltu
r
e
m
ed
ia
aim
s
to
p
r
o
v
id
e
a
b
alan
ce
o
f
n
u
tr
ien
t
m
ix
tu
r
es
n
ee
d
ed
b
y
m
icr
o
o
r
g
an
is
m
s
b
y
p
r
o
v
id
i
n
g
an
ar
tific
ial
en
v
ir
o
n
m
en
t
as
a
s
im
u
latio
n
o
f
n
atu
r
al
co
n
d
itio
n
s
th
at
ar
e
im
p
o
r
tan
t f
o
r
g
r
o
w
th
.
Mic
r
o
o
r
g
an
is
m
s
ar
r
an
g
e
t
h
eir
ce
ll
co
m
p
o
n
e
n
ts
b
y
ass
em
b
lin
g
s
m
all
m
o
lec
u
les
u
tili
zin
g
n
u
tr
ien
ts
in
th
e
g
r
o
wth
m
ed
i
u
m
.
Dif
f
er
en
t
m
icr
o
o
r
g
a
n
is
m
s
r
eq
u
ir
e
d
if
f
er
en
t n
u
tr
itio
n
al
m
ate
r
ials
.
T
h
er
ef
o
r
e,
th
e
r
e
ar
e
c
u
ltu
r
e
m
ed
ia
in
v
ar
y
in
g
s
h
a
p
es
an
d
c
o
m
p
o
s
itio
n
s
ad
ap
ted
to
th
e
ty
p
e
o
f
m
icr
o
o
r
g
an
is
m
s
b
ein
g
cu
ltu
r
ed
[
1
2
]
,
[
1
3
]
.
Ma
cCo
n
k
ey
m
ed
iu
m
is
a
s
elec
tiv
e
an
d
d
if
f
er
en
tial
m
ed
i
u
m
f
o
r
th
e
g
r
o
wth
o
f
g
r
am
-
n
e
g
ativ
e
b
ac
ter
ia
b
ased
o
n
th
e
b
ac
ter
ia'
s
ab
ilit
y
to
b
r
ea
k
d
o
w
n
lacto
s
e.
T
h
e
m
ain
co
m
p
o
n
e
n
ts
o
f
Ma
cCo
n
k
ey
m
e
d
iu
m
in
clu
d
e
cr
y
s
tal
v
io
let
d
y
es,
b
ile
s
alts
,
l
ac
to
s
e,
an
d
n
eu
tr
al
r
ed
(
p
H
in
d
icato
r
)
.
C
r
y
s
tal
v
io
let
d
y
es a
n
d
b
ile
s
alts
s
to
p
th
e
g
r
o
wth
o
f
g
r
am
-
p
o
s
itiv
e
b
ac
t
er
ia.
Ma
cCo
n
k
ey
m
ed
iu
m
co
n
tain
s
a
p
H
in
d
icato
r
th
at
tu
r
n
s
p
in
k
u
n
d
e
r
ac
id
ic
co
n
d
itio
n
s
.
B
ac
ter
ia
th
at
ca
n
f
er
m
en
t
lacto
s
e
will
p
r
o
d
u
ce
o
r
g
an
ic
ac
id
s
,
esp
ec
ially
lacti
c
ac
id
,
wh
ich
ca
n
lo
wer
p
H.
T
h
e
p
r
esen
ce
o
f
n
e
u
tr
al
r
ed
as
a
p
H
in
d
icato
r
wi
ll
ch
an
g
e
f
r
o
m
f
a
d
ed
w
h
ite
to
b
r
ig
h
t
r
ed
o
r
p
in
k
wh
en
th
e
p
H
is
less
th
an
6
.
8
.
T
h
u
s
,
p
i
n
k
co
lo
n
ies
will
f
o
r
m
o
n
lacto
s
e
f
e
r
m
en
ter
b
ac
ter
ia
,
wh
ile
n
o
n
-
lacto
s
e
f
er
m
en
ter
s
will f
o
r
m
o
p
aq
u
e
wh
ite
co
lo
n
ies
.
Path
o
g
en
ic
b
ac
ter
ia,
esp
ec
ially
b
ac
ter
ia
th
at
ca
u
s
e
g
astro
en
ter
itis
s
u
ch
as
E
s
ch
erich
ia
an
d
S
a
lmo
n
ella
,
ar
e
wid
el
y
d
is
tin
g
u
is
h
e
d
u
s
in
g
th
is
m
ed
i
u
m
[
1
4
]
.
Nu
t
r
i
en
t
a
g
a
r
is
o
n
e
o
f
t
h
e
c
o
m
p
le
x
m
ed
ia
.
P
er
l
ite
r
,
a
g
a
r
n
u
t
r
ie
n
ts
co
n
t
ai
n
b
ee
f
e
x
t
r
ac
t
(
3
g
)
,
p
ep
to
n
e
(5
g
)
,
s
o
d
i
u
m
ch
lo
r
i
d
e
(
5
g
)
,
a
n
d
a
g
a
r
(
1
5
g
)
.
T
h
e
m
ai
n
c
o
m
p
o
n
en
ts
o
f
n
u
tr
i
e
n
t
a
g
a
r
m
e
d
i
a
ar
e
c
ar
b
o
h
y
d
r
at
es
an
d
p
r
o
t
ei
n
s
f
o
u
n
d
i
n
m
e
at
e
x
t
r
a
cts
a
n
d
p
ep
to
n
es.
T
h
ese
n
u
t
r
ien
ts
p
r
o
v
i
d
e
a
s
o
u
r
ce
o
f
n
i
tr
o
g
e
n
a
n
d
c
ar
b
o
n
,
as
well
as
v
ita
m
i
n
s
t
o
a
id
in
b
ac
ter
ial
g
r
o
wt
h
[
1
5
]
.
T
h
e
n
u
tr
i
e
n
t
c
o
m
p
o
s
iti
o
n
o
f
a
g
ar
an
d
n
u
t
r
ie
n
t
b
r
o
t
h
ca
n
b
e
r
e
p
l
ac
e
d
wit
h
al
te
r
n
at
iv
e
in
g
r
e
d
ie
n
ts
w
it
h
s
i
m
il
a
r
n
u
t
r
iti
o
n
a
l
co
n
t
en
t t
o
s
u
p
p
o
r
t
b
a
ct
er
i
al
g
r
o
wt
h
[
1
6
]
.
T
h
e
S
a
lmo
n
ella
typ
h
i
is
o
late
t
o
b
e
id
en
tifie
d
n
ee
d
s
to
b
e
is
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b
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Hea
lth
Sci
,
Vo
l.
13
,
No
.
3
,
Sep
tem
b
er
20
24
:
48
-
55
50
p
r
o
ce
s
s
is
n
ec
ess
ar
y
th
at
in
v
o
lv
es
u
s
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tim
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f
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latio
n
s
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iv
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e
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ac
te
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ia
wh
ile
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ed
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cin
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r
esen
ce
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m
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etin
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cter
ia.
Nu
tr
ien
t
a
g
ar
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e
x
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le
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n
o
n
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s
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m
en
t
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et
h
o
d
u
s
ed
f
o
r
is
o
latin
g
S
a
lmo
n
ella
[
1
7
]
.
I
D
b
r
o
th
is
a
b
r
o
th
u
s
ed
as
a
m
an
u
f
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ial
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au
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atic
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tib
io
tic
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u
s
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p
tib
ilit
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test
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g
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th
e
Ph
o
en
ix
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y
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tem
.
Po
s
itiv
e
cu
ltu
r
es
o
f
g
r
o
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g
b
ac
ter
ia
n
e
ed
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b
e
test
ed
f
o
r
an
tim
icr
o
b
ial
s
u
s
ce
p
tib
ilit
y
test
in
g
(
AST
)
r
esis
tan
ce
.
AST
is
a
cr
itica
l
p
r
o
ce
s
s
in
cl
in
ical
m
icr
o
b
io
lo
g
y
to
ev
alu
ate
an
tib
io
tic
ef
f
ec
tiv
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e
s
s
.
I
t id
en
tifie
s
wh
en
b
ac
ter
ia
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ec
o
m
e
less
s
u
s
ce
p
tib
le
o
r
r
esis
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t to
an
tib
io
tics
,
r
ed
u
cin
g
th
eir
a
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ilit
y
to
elim
in
ate
b
ac
ter
ia
o
r
in
h
ib
it
th
eir
g
r
o
wth
[
1
8
]
.
Ph
o
en
i
x
'
s
au
to
m
atic
AST
r
es
is
tan
c
e
test
was
p
er
f
o
r
m
ed
b
y
m
ak
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g
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lu
m
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p
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th
i
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th
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m
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n
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f
0
.
5
-
0
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6
Mc
Far
lan
d
.
T
h
e
b
ac
ter
ial
s
u
s
p
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n
s
io
n
f
r
o
m
I
D
B
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is
tr
an
s
f
er
r
e
d
to
AST
b
r
o
th
a
n
d
co
n
tin
u
ed
wit
h
au
to
m
ate
d
test
in
g
[
1
9
]
.
T
h
e
in
o
cu
lu
m
s
u
s
p
en
s
io
n
o
n
I
D
B
r
o
th
ca
n
b
e
u
s
ed
as a
n
ad
v
a
n
ce
d
test
in
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m
ater
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I
n
th
is
s
tu
d
y
,
test
in
g
will
b
e
ca
r
r
ied
o
u
t
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n
t
h
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ef
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ctiv
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tr
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M
E
T
H
O
D
I
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tu
d
y
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u
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S
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lmo
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lated
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2
0
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2
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s
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h
e
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en
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ch
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as
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n
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u
cted
at
th
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Dep
ar
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en
t
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Me
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ical
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ab
o
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ato
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ly
tech
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c
Min
is
tr
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f
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Su
r
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ay
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th
e
I
n
s
titu
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r
o
p
ical
Dis
ea
s
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o
m
Ap
r
i
l 2
0
2
3
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2
0
2
3
,
u
s
in
g
th
r
ee
s
am
p
les o
f
S
a
lmo
n
ella
typ
h
i
is
o
lates f
r
o
m
E
ast
J
av
a
with
th
r
ee
r
ep
licatio
n
s
.
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h
is
s
tu
d
y
was
ap
p
r
o
v
ed
b
y
th
e
Hea
lth
R
esear
ch
E
th
ics
C
o
m
m
ittee
at
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ly
tech
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ic
Min
is
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o
f
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lth
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r
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ay
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with
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0
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ltek
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ated
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.
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a
k
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edia
Me
d
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c
c
u
r
s
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t
h
e
B
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ct
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o
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ab
o
r
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to
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y
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t
h
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p
a
r
tm
e
n
t
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Me
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ic
al
L
a
b
o
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o
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y
T
ec
h
n
o
l
o
g
y
at
th
e
P
o
l
y
t
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h
n
i
c
Mi
n
is
tr
y
o
f
H
ea
l
th
i
n
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u
r
a
b
a
y
a
.
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n
t
h
is
r
esea
r
ch
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w
e
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t
iliz
ed
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cCo
n
k
e
y
m
e
d
i
u
m
C
M0
1
1
5
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I
D
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d
n
u
t
r
ie
n
t
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g
a
r
C
M
0
0
0
3
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OXOI
D)
.
T
h
e
m
e
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ia
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p
r
e
p
ar
ed
f
o
ll
o
w
in
g
th
e
OXOI
D
p
r
o
ce
d
u
r
es.
T
o
c
r
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at
e
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cC
o
n
k
e
y
m
e
d
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u
m
C
M
0
1
1
5
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D
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ed
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,
4
9
.
5
3
g
r
a
m
s
o
f
m
e
d
i
a
p
o
w
d
e
r
,
co
n
s
is
ti
n
g
o
f
p
ep
t
o
n
e
(
1
7
g
)
,
p
r
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te
o
s
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p
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t
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e
(
3
g
)
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l
ac
t
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s
e
m
o
n
o
h
y
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r
ate
(
1
0
g
)
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ile
s
al
ts
(
1
.
5
g
)
,
s
o
d
i
u
m
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h
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o
r
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(
5
g
)
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n
e
u
t
r
a
l
r
e
d
(
3
0
m
g
)
,
c
r
y
s
tal
v
i
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le
t
(
1
m
g
)
,
a
n
d
ag
ar
(
1
3
.
5
g
)
,
is
d
is
s
o
l
v
e
d
i
n
o
n
e
li
te
r
o
f
a
q
u
a
d
est
.
T
h
e
m
e
d
ia
s
h
o
u
l
d
b
e
d
is
s
o
l
v
e
d
w
h
i
le
s
ti
r
r
in
g
a
t
8
0
º
C
.
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f
t
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d
is
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lv
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t
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a
n
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ad
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s
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it
t
o
t
h
e
f
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n
al
p
H
o
f
N
A
m
e
d
ia
,
w
h
ic
h
is
7
.
1
±
0
.
2
.
Fi
n
al
ly
,
w
r
a
p
t
h
e
m
e
d
ia
a
n
d
s
te
r
il
iz
e
u
s
i
n
g
a
1
2
1
º
C
a
u
t
o
cla
v
e
f
o
r
1
5
m
i
n
u
tes
.
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o
c
r
e
at
e
n
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t
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n
t
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ar
C
M
0
0
0
3
(
OX
OI
D
)
m
e
d
ia
,
2
8
g
r
am
s
o
f
m
e
d
ia
p
o
w
d
e
r
,
c
o
n
s
is
t
in
g
o
f
p
e
p
t
o
n
e
(
5
g
)
,
y
ea
s
t
e
x
tr
ac
t
(
2
g
)
,
s
o
d
i
u
m
c
h
l
o
r
i
d
e
(
5
g
)
,
b
e
ef
e
x
t
r
ac
t
(
1
g
)
,
a
n
d
a
g
a
r
(
1
5
g
)
,
is
d
is
s
o
l
v
e
d
i
n
o
n
e
li
te
r
o
f
aq
u
a
d
est.
T
h
e
m
e
d
ia
s
h
o
u
l
d
b
e
d
is
s
o
l
v
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d
w
h
il
e
s
ti
r
r
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n
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at
8
0
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f
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ch
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p
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n
d
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d
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u
s
t
i
t
t
o
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e
f
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n
a
l
p
H
o
f
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m
e
d
i
a,
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i
ch
is
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.
4
±
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.
2
.
Fi
n
al
ly
,
w
r
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p
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h
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m
e
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a
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n
d
s
te
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liz
e
u
s
in
g
a
1
2
1
º
C
a
u
t
o
c
la
v
e
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o
r
1
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m
i
n
u
tes
.
2
.
2
.
Sa
lm
o
nella
typ
hi
is
o
la
t
io
n
Fo
r
th
is
s
tu
d
y
,
two
ty
p
es
o
f
m
ed
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-
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cCo
n
k
ey
m
ed
iu
m
an
d
I
D
B
r
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th
-
wer
e
u
s
ed
to
in
o
cu
late
S
a
lmo
n
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h
i
b
ac
ter
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T
h
e
b
ac
ter
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e
g
r
o
wn
o
n
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cCo
n
k
ey
m
ed
iu
m
u
s
in
g
th
e
s
t
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k
tech
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iq
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e
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d
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h
o
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r
s
.
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n
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D
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r
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th
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th
e
b
ac
te
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e
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o
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lev
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f
0
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5
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6
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Far
lan
d
.
Af
ter
g
r
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wth
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S
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lmo
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f
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al
s
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g
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o
n
Nu
tr
ien
t
Ag
ar
a
s
a
s
u
p
p
lem
en
ta
r
y
m
ed
iu
m
.
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o
o
b
tain
a
p
u
r
e
cu
l
tu
r
e
o
f
th
e
b
ac
ter
ia
f
o
r
f
u
r
th
e
r
test
in
g
,
th
e
s
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p
les
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s
t
r
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k
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tr
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Ag
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t
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b
ated
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f
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.
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.
O
M
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lm
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C
f
o
r
2
0
m
in
.
C
en
tr
if
u
g
es
r
etu
r
n
is
o
lates
at
4
º
C
to
1
5
,
0
0
0
r
p
m
f
o
r
3
0
m
in
u
tes.
T
h
e
p
ellets
wer
e
r
es
u
s
p
en
d
ed
with
5
0
0
μ
L
PB
S.
Sto
r
e
at
-
20
º
C
u
n
til
r
ea
d
y
to
u
s
e.
Evaluation Warning : The document was created with Spire.PDF for Python.
I
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2252
-
8
8
0
6
Th
e
effec
tiven
ess
o
f u
s
in
g
I
D
b
r
o
th
in
id
en
tifyin
g
th
e
o
u
ter me
mb
r
a
n
e
p
r
o
tein
…
(
P
esta
r
ia
ti
)
51
2
.
4
.
P
r
o
t
ein c
o
nte
nt
ca
lcula
t
io
n
Pro
tein
lev
els
wer
e
m
ea
s
u
r
e
d
u
s
in
g
th
er
m
o
s
cien
tific
n
an
o
d
r
o
p
at
a
wav
elen
g
th
o
f
2
8
0
n
m
[
2
1
]
.
SDS
-
PAGE
T
o
ex
tr
ac
t
th
e
S
a
lmo
n
ella
typ
h
i
OM
P
an
tig
en
,
we
u
s
ed
SDS
-
P
AGE
with
a
1
2
%
s
ep
ar
atin
g
g
el
co
n
ce
n
tr
atio
n
an
d
a
4
%
s
tack
in
g
g
el.
W
e
m
ix
e
d
th
e
cr
u
d
e
s
am
p
les
an
d
lo
a
d
in
g
b
u
f
f
er
s
i
n
m
icr
o
ce
n
tr
if
u
g
e
tu
b
es
at
a
r
atio
o
f
2
:1
a
n
d
h
o
m
o
g
en
ized
th
em
u
s
in
g
s
p
in
d
o
wn
.
Nex
t,
we
h
ea
te
d
th
e
s
am
p
le
f
o
r
5
m
in
u
tes
b
etwe
en
9
0
ºC
-
95
º
C
.
W
e
p
r
e
p
ar
ed
t
h
e
g
el
in
a
ch
a
m
b
er
with
a
r
u
n
n
i
n
g
b
u
f
f
er
a
n
d
f
illed
th
e
av
ailab
le
wells
with
2
0
μ
L
o
f
p
r
ep
a
r
ed
s
am
p
les.
W
e
al
s
o
f
illed
o
n
e
well
with
1
0
μ
L
o
f
p
r
o
tein
m
ar
k
er
s
.
W
e
r
an
elec
tr
o
p
h
o
r
esis
at
1
0
0
V
4
0
m
A
f
o
r
9
0
m
in
u
tes,
u
n
til
th
e
s
am
p
le
alm
o
s
t
r
ea
ch
ed
th
e
g
el
b
o
tto
m
.
Fin
ally
,
we
s
tain
ed
th
e
g
el
u
s
in
g
c
o
o
m
ass
ie
b
r
illi
an
t b
lu
e
R
-
2
5
0
(
B
io
-
R
ad
)
.
3.
RE
SU
L
T
S AN
D
D
I
SCU
SS
I
O
N
Ma
cCo
n
k
ey
m
ed
iu
m
is
a
ty
p
e
o
f
b
ac
ter
ial
g
r
o
wth
m
ed
iu
m
th
at
is
b
o
th
s
elec
tiv
e
an
d
d
if
f
er
en
tial
,
m
ea
n
in
g
it
is
u
s
ed
to
id
en
tify
an
d
is
o
late
g
r
am
-
n
eg
ativ
e
b
ac
ter
ia.
T
h
is
m
ed
ia
h
as
a
c
o
m
p
o
s
itio
n
o
f
b
ile
s
alts
as
in
h
ib
ito
r
s
o
f
g
r
a
m
-
p
o
s
iti
v
e
b
ac
ter
ia,
n
e
u
tr
al
r
ed
d
y
e
as
an
in
d
icato
r
o
f
p
H,
lac
to
s
e,
an
d
p
ep
t
o
n
e
.
S
a
lmo
n
ella
typ
h
i
ap
p
ea
r
s
as
w
h
ite
co
lo
n
ies
t
h
at
d
o
n
o
t
ca
u
s
e
an
y
ch
an
g
es
to
th
e
p
H
in
th
e
ab
s
en
ce
o
f
lacto
s
e
f
er
m
en
tatio
n
.
Ma
cCo
n
k
e
y
is
a
co
m
m
o
n
ly
u
s
ed
g
r
o
wth
m
e
d
i
u
m
th
at
s
elec
tiv
ely
p
r
o
m
o
tes
th
e
g
r
o
wth
o
f
g
r
am
-
n
eg
ativ
e
b
ac
ter
ia
a
n
d
d
is
tin
g
u
is
h
es
th
em
b
ased
o
n
th
eir
f
e
r
m
en
tatio
n
p
r
o
f
ile.
T
h
is
m
ed
i
a
is
wid
ely
u
s
ed
to
d
if
f
er
en
tiate
p
at
h
o
g
e
n
ic
b
ac
ter
ia,
esp
ec
ially
in
s
p
ec
ies th
at
ca
u
s
e
g
astro
en
ter
itis
[
1
4
]
,
[
2
2
]
.
I
D
B
r
o
th
(
B
D
Ph
o
en
ix
I
D
B
r
o
th
)
is
a
s
o
lu
tio
n
u
s
ed
to
cr
ea
te
b
ac
ter
ial
s
u
s
p
en
s
io
n
s
f
o
r
u
s
e
in
Ph
o
en
ix
'
s
au
to
m
ated
s
u
s
ce
p
tib
ilit
y
id
en
tific
atio
n
an
d
test
in
g
s
y
s
tem
.
I
t
co
n
tain
s
p
o
tass
iu
m
ch
lo
r
id
e,
ca
lciu
m
ch
lo
r
id
e,
tr
icin
e
g
l
y
cin
e,
an
d
p
o
ly
s
o
r
b
ate
8
0
in
a
o
n
e
-
liter
s
o
lu
tio
n
.
T
h
e
am
o
u
n
t
o
f
b
ac
t
er
ial
s
u
s
p
en
s
io
n
is
d
eter
m
in
ed
u
s
in
g
Mc
Far
lan
d
s
tan
d
ar
d
s
.
Gly
cin
e
is
o
n
e
o
f
th
e
ess
en
tial
am
in
o
ac
id
s
an
d
is
a
s
o
u
r
ce
o
f
co
n
d
en
s
ed
n
itro
g
en
[
2
3
]
.
T
r
is
-
tr
icin
e
h
as
b
ee
n
f
o
u
n
d
to
b
e
a
s
u
p
er
io
r
b
u
f
f
e
r
f
o
r
s
ep
ar
ati
n
g
s
er
u
m
p
r
o
tein
s
an
d
is
co
m
m
o
n
ly
u
s
ed
in
co
n
ju
n
ct
io
n
with
g
ly
cin
e
to
s
tab
ilize
p
H.
I
D
B
r
o
th
co
n
tain
s
n
o
t
o
n
ly
tr
icin
e
an
d
g
ly
cin
e,
b
u
t
also
p
o
ly
s
o
r
b
ate
8
0
.
T
r
i
cin
e
is
a
r
ec
o
m
m
en
d
ed
m
et
h
o
d
f
o
r
co
n
tr
o
llin
g
p
H,
as
it
clo
s
ely
r
esem
b
les
p
h
y
s
io
lo
g
ical
s
o
lu
tio
n
s
[
2
4
]
,
[
2
5
]
.
T
h
er
ap
eu
tic
p
r
o
tein
f
o
r
m
u
latio
n
s
u
s
e
Po
ly
s
o
r
b
ate
8
0
as
a
s
u
r
f
ac
tan
t
to
p
r
ev
en
t
ad
s
o
r
p
tio
n
an
d
p
r
o
m
o
te
p
r
o
tein
ag
g
r
eg
atio
n
[
2
6
]
.
Po
ly
s
o
r
b
ate
im
p
ac
ts
S
a
lmo
n
ella
typ
h
i
b
y
s
tab
ilizin
g
b
ac
ter
ial
ce
lls
an
d
r
ed
u
cin
g
th
eir
clu
m
p
in
g
,
wh
ich
ca
n
in
ter
f
e
r
e
with
m
ea
s
u
r
in
g
p
r
o
tein
lev
els.
Po
ly
s
o
r
b
ate
ca
n
also
in
f
lu
en
c
e
th
e
in
teg
r
ity
o
f
th
e
b
ac
ter
ial
ce
ll
m
em
b
r
a
n
e
an
d
alter
p
r
o
t
ein
s
tr
u
ctu
r
e
with
i
n
th
e
ce
lls
[
2
7
]
.
Nu
tr
ien
t
Ag
ar
is
a
co
m
m
o
n
ly
u
s
ed
n
o
n
-
s
elec
tiv
e
m
ed
ia
f
o
r
r
ap
id
ly
g
r
o
win
g
v
ar
io
u
s
b
ac
ter
ial
s
p
ec
ies
f
o
u
n
d
in
wate
r
,
waste,
an
d
f
e
ce
s
.
I
t
co
n
s
is
ts
o
f
p
ep
t
o
n
e,
b
e
ef
ex
tr
ac
t,
a
n
d
a
g
ar
,
p
r
o
v
id
in
g
ess
en
tial
n
u
tr
ien
ts
f
o
r
b
ac
ter
ial
g
r
o
wth
.
T
h
is
m
ak
es
it
an
ex
ce
llen
t
ch
o
ice
f
o
r
b
ac
ter
ial
id
en
tific
atio
n
,
esp
ec
ially
f
o
r
cu
ltiv
atin
g
lar
g
e
q
u
an
titi
es
o
f
Salm
o
n
ella
b
ac
ter
ia
[
2
8
]
.
I
n
a
s
tu
d
y
c
o
n
d
u
cte
d
b
y
Kh
o
s
r
av
i
(
2
0
2
2
)
,
im
m
u
n
o
m
a
g
n
etic
b
ea
d
s
wer
e
d
ev
el
o
p
ed
t
o
is
o
late
Salm
o
n
ella
b
ac
ter
ia
f
r
o
m
co
n
tam
in
ated
m
ilk
s
am
p
l
es.
T
h
e
r
esear
ch
er
s
s
u
cc
ess
f
u
lly
d
etec
ted
at
least
3
×1
0
4
C
FU/m
L
o
f
b
ac
ter
ia
i
n
th
e
p
o
llu
ted
s
am
p
les
u
s
in
g
n
u
tr
ien
t
ag
ar
m
e
d
ia
[
2
9
]
.
S
a
lmo
n
ella
typ
h
i
is
o
la
te
is
e
x
tr
ac
ted
u
s
in
g
s
o
n
icatio
n
tec
h
n
iq
u
es
b
y
u
tili
zin
g
u
ltra
s
o
n
i
c
en
er
g
y
to
m
o
v
e
th
e
co
n
s
titu
en
t
p
ar
ticles
o
f
th
e
ce
ll
m
e
m
b
r
an
e
s
o
th
at
th
e
p
r
o
tein
co
m
p
o
n
e
n
ts
ca
n
b
e
r
elea
s
ed
.
OM
P
ex
tr
ac
tio
n
o
f
S
a
lmo
n
ella
typ
h
i
in
th
is
s
tu
d
y
u
s
ed
a
s
o
n
icatio
n
tech
n
iq
u
e
with
a
f
r
eq
u
e
n
c
y
o
f
3
5
k
Hz
f
o
r
3
0
s
ec
o
n
d
s
an
d
r
ep
ea
ted
f
o
r
1
5
cy
cles
with
a
g
ap
o
f
3
0
s
ec
o
n
d
s
.
Af
ter
th
e
s
o
n
icatio
n
p
r
o
ce
s
s
,
p
h
o
s
p
h
ate
-
b
u
f
f
er
e
d
s
alin
e
(
PB
S
)
s
u
r
f
ac
tan
t
an
d
T
r
ito
n
-
X
ar
e
ad
d
e
d
to
ass
is
t
in
ly
s
in
g
th
e
b
ac
ter
ial
ce
ll
s
u
r
f
ac
e
s
o
th
at
OM
P
ex
tr
ac
t
is
o
b
tain
ed
to
b
e
an
aly
ze
d
.
PB
S
is
a
wate
r
-
b
ased
s
alt
s
o
lu
tio
n
u
s
ed
to
m
a
in
tain
th
e
p
H
an
d
o
s
m
o
lar
ity
f
o
r
ce
lls
,
tis
s
u
es,
a
n
d
b
io
l
o
g
ical
s
am
p
les
[
3
0
]
.
T
h
e
p
r
o
tein
co
n
ten
t
o
f
OM
P
S
a
lmo
n
ella
typ
h
i
ex
tr
ac
t
wa
s
m
ea
s
u
r
ed
u
s
in
g
a
T
h
er
m
o
Scien
tific
Nan
o
d
r
o
p
at
a
wav
elen
g
th
o
f
2
8
0
n
m
.
T
h
er
m
o
Scien
tific
n
an
o
d
r
o
p
u
tili
ze
s
a
UV
-
Vis
s
p
ec
tr
o
p
h
o
t
o
m
eter
to
m
ea
s
u
r
e
ab
s
o
r
b
an
ce
.
Pu
r
if
ied
p
r
o
tein
s
ab
s
o
r
b
lig
h
t
at
a
p
ea
k
o
f
2
8
0
n
m
[
2
1
]
.
T
a
b
le
1
s
h
o
ws
th
at
th
e
p
r
o
tein
lev
els in
S
a
lmo
n
ella
typ
h
i
OM
P e
x
tr
ac
t o
b
tain
ed
f
r
o
m
I
D
B
r
o
th
cu
ltu
r
e
s
am
p
les r
an
g
ed
f
r
o
m
8
.
3
1
0
m
g
/m
L
to
8
.
9
8
5
m
g
/m
L
,
wh
ile
s
am
p
les
f
r
o
m
Ma
cCo
n
k
ey
cu
ltu
r
es
r
an
g
ed
f
r
o
m
1
.
1
0
5
m
g
/m
L
to
5
.
4
7
1
m
g
/m
L
.
T
h
e
d
if
f
er
en
ce
i
n
b
ac
ter
ial
g
r
o
wth
b
etwe
en
s
o
lid
an
d
liq
u
id
c
u
ltu
r
e
m
ed
ia
m
a
y
b
e
d
u
e
to
d
if
f
er
en
ce
s
in
n
u
tr
ie
n
t
av
ailab
ilit
y
.
B
ef
o
r
e
p
r
o
tein
elec
tr
o
p
h
o
r
es
is
,
OM
P
ex
tr
ac
t
was
p
r
ep
ar
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-
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-
PAGE
.
T
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I
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2
2
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I
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b
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Hea
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Sci
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3
,
Sep
tem
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20
24
:
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-
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52
p
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r
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Gel
An
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9
.
1
ap
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licatio
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h
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SDS
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PA
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1
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Evaluation Warning : The document was created with Spire.PDF for Python.
I
n
t J Pu
b
lic
Hea
lth
Sci
I
SS
N:
2252
-
8
8
0
6
Th
e
effec
tiven
ess
o
f u
s
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I
D
b
r
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th
in
id
en
tifyin
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ter me
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r
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p
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…
(
P
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r
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ti
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53
T
h
e
m
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lec
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lar
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h
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p
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b
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d
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o
r
m
ed
a
f
ter
O
MP
s
ep
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n
u
s
in
g
SDS
-
PAGE
was
m
ea
s
u
r
ed
u
s
in
g
Gel
An
aly
ze
r
1
9
.
1
s
o
f
twar
e,
f
o
llo
win
g
s
t
ain
in
g
with
c
o
o
m
ass
ie
b
r
illi
a
n
t
b
lu
e
.
Fro
m
t
h
e
m
ea
s
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r
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en
t
r
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h
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r
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m
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d
etails
in
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ab
le
2
.
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h
e
d
if
f
er
e
n
ce
in
t
h
e
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m
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e
r
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d
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d
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tim
al,
p
r
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b
a
n
d
s
ca
n
n
o
t
b
e
r
ea
d
,
an
d
m
o
lecu
lar
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h
t
ca
n
n
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t
b
e
id
en
tifie
d
[
3
2
]
.
T
h
e
p
r
o
tein
b
an
d
f
o
r
m
e
d
in
SDS
-
PAGE
ca
n
b
e
d
eter
m
in
ed
f
r
o
m
th
e
r
esu
lts
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f
s
am
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le
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x
tr
ac
tio
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ca
r
r
ie
d
o
u
t
p
r
ev
io
u
s
ly
,
c
o
n
s
id
er
in
g
th
at
in
th
is
s
tu
d
y
s
am
p
le
ex
tr
ac
tio
n
with
th
e
s
am
e
m
et
h
o
d
,
th
e
d
if
f
e
r
en
ce
in
r
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ca
n
b
e
ca
u
s
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y
th
e
p
r
o
tein
le
v
els
co
n
tain
e
d
.
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h
e
d
is
ad
v
a
n
tag
e
o
f
th
is
SDS
-
PAGE
m
eth
o
d
is
th
at
p
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a
n
d
v
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d
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n
e
m
an
u
ally
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n
th
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u
g
h
m
o
lec
u
lar
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h
t
ca
lcu
latio
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s
ar
e
d
o
n
e
au
to
m
ati
ca
lly
u
s
in
g
Gel
An
aly
ze
r
1
9
.
1
.
I
t
is
co
m
m
o
n
to
o
b
s
er
v
e
d
if
f
er
en
ce
s
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etwe
en
th
e
ac
tu
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o
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lar
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t
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f
p
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s
o
n
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DS
-
PAGE
g
els
an
d
th
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ex
p
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ted
s
ize.
T
h
is
d
if
f
er
en
ce
ca
n
ar
is
e
d
u
e
to
ch
em
ical
m
o
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f
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n
s
s
u
ch
as
g
ly
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s
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latio
n
an
d
u
b
i
q
u
itin
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,
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ich
c
an
s
ig
n
if
ican
tly
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ec
t
th
eir
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o
b
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y
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n
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e
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el.
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t'
s
wo
r
th
n
o
tin
g
th
at
p
h
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s
p
h
o
r
y
latio
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ca
n
s
o
m
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es
ca
u
s
e
s
lig
h
t
v
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n
s
in
SDS
-
PAGE
g
els.
Ho
w
ev
er
,
wh
en
h
y
p
er
-
p
h
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s
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h
o
r
y
latio
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o
cc
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s
,
it
s
h
o
ws
u
p
as
s
lo
wer
s
tain
s
.
T
h
er
ef
o
r
e,
it
is
im
p
o
r
ta
n
t
to
ex
am
in
e
wh
eth
er
th
e
d
if
f
er
en
ce
in
s
ize
is
d
u
e
to
ch
em
ical
m
o
d
if
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s
o
r
s
p
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if
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f
ea
tu
r
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s
u
c
h
as
th
e
p
r
o
tein
'
s
am
in
o
ac
i
d
co
m
p
o
s
itio
n
[
3
1
]
.
4.
CO
NCLU
SI
O
N
R
esear
ch
h
as
s
h
o
wn
th
at
OM
P
S
a
lmo
n
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h
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e
x
tr
ac
t
g
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o
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Nu
tr
ie
n
t
Ag
ar
f
r
o
m
I
D
B
r
o
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co
n
tain
s
h
ig
h
er
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r
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lev
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an
S
a
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n
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i
cu
ltu
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e
d
f
r
o
m
Ma
cCo
n
k
ey
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ed
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m
.
T
h
is
v
ar
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n
is
d
u
e
to
en
v
ir
o
n
m
en
tal
co
n
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itio
n
s
an
d
n
u
tr
ie
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t
ab
s
o
r
p
tio
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with
in
th
e
m
ed
ia.
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n
g
I
D
B
r
o
t
h
b
ef
o
r
e
g
r
o
win
g
o
n
Nu
tr
ien
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Ag
ar
s
tab
ilizes
th
e
b
ac
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d
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Evaluation Warning : The document was created with Spire.PDF for Python.