The effects of ozone, chlorine dioxide, cetylpyridinium chloride and trisodium phosphate as multiple antimicrobial interventions on microbiological, instrumental color, and sensory color and odor characteristics of ground beef


Pohlman, F.; Stivarius, M.; Mcelyea, K.; Johnson, Z.; Johnson, M.

Meat Science 61(3): 307-313

2002


The impact of multiple antimicrobial interventions on ground beef microbial, color and sensory characteristics was studied. For this, beef trimmings were inoculated with Escherichia coli (EC) and Salmonella typhimurium (ST) then treated with either (1) 1% ozonated water followed by 5% acetic acid (OA), (2) 1% ozonated water followed by 0.5% cetylpyridinium chloride (OC), (3) 200 ppm chlorine dioxide followed by 10% trisodium phosphate (CT) or (4) control (C). Trimmings were ground, packaged and sampled at 0, 1, 2, 3 and 7 days of display for EC, ST, coliforms (CO), aerobic plate count (APC), instrumental color and sensory color and odor characteristics. The OA and OC treatments reduced (P < 0.05) all bacterial types evaluated, while CT reduced (P < 0.05) EC, CO and APC. The CT treatment was redder (P < 0.05) in overall color than C, and there was no difference (P > 0.05) in beef odor or off odor between OC, CT or C treatments.

MEAT
SCIENCE
ELSEVIER
Meat
Science
61
(2002)
307-313
www.elsevier.com/locate/meatsci
The
effects
of
ozone,
chlorine
dioxide,
cetylpyridinium
chloride
and
trisodium
phosphate
as
multiple
antimicrobial
interventions
on
microbiological,
instrumental
color,
and
sensory
color
and
odor
characteristics
of
ground
beef
F.W.
Pohlmana
,
*,
M.R.
Stivariusb,
K.S.
McElyeaa,
Z.B.
Johnsona,
M.G.
Johnsonc
'Department
of
Animal
Science,
University
of
Arkansas,
Fayetteville,
AR
72701,
USA
bGriffith
Laboratories,
Griffith
Center,
Alsip,
IL
60658,
USA
'Department
of
Food
Science,
University
of
Arkansas,
Fayetteville,
AR
72701,
USA
Received
26
March
2001;
received
in
revised
form
20
September
2001;
accepted
20
September
2001
Abstract
The
impact
of
multiple
antimicrobial
interventions
on
ground
beef
microbial,
color
and
sensory
characteristics
was
studied.
For
this,
beef
trimmings
were
inoculated
with
Escherichia
coli
(EC)
and
Salmonella
typhimurium
(ST)
then
treated
with
either
(1)
1%
ozonated
water
followed
by
5%
acetic
acid
(OA),
(2)
1%
ozonated
water
followed
by
0.5%
cetylpyridinium
chloride
(OC),
(3)
200
ppm
chlorine
dioxide
followed
by
10%
trisodium
phosphate
(CT)
or
(4)
control
(C).
Trimmings
were
ground,
packaged
and
sampled
at
0,
1,
2,
3
and
7
days
of
display
for
EC,
ST,
coliforms
(CO),
aerobic
plate
count
(APC),
instrumental
color
and
sensory
color
and
odor
characteristics.
The
OA
and
OC
treatments
reduced
(P
<
0.05)
all
bacterial
types
evaluated,
while
CT
reduced
(P
<
0.05)
EC,
CO
and
APC.
The
CT
treatment
was
redder
(P
<
0.05)
in
overall
color
than
C,
and
there
was
no
difference
(P>
0.05)
in
beef
odor
or
off
odor
between
OC,
CT
or
C
treatments.
©
2002
Elsevier
Science
Ltd.
All
rights
reserved.
Keywords:
Ground
beef;
Chlorine
dioxide;
Acetic
acid;
Ozone;
Cetylpyridinium
chloride;
Meat
color
1.
Introduction
Antimicrobial
intervention
treatment
of
carcasses
and
meat
tissues
have
been
investigated
and
have
ranged
from
water
washing
and
steam
pasteurization
(Phebus
et
al.,
1997;
Reagan
et
al.,
1996)
to
organic
acids,
alka-
line
phosphates
and
other
novel
compounds
(Kim
&
Slavik,
1994;
Kochevar,
Sofos,
LeValley,
&
Smith,
1997).
Recently,
research
has
been
conducted
using
the
concept
of
hurdle
technology
(Ellebracht,
Castillo,
Lucia,
Miller,
&
Acuff,
1999;
Phebus
et
al.,
1997).
Hur-
dle
technology
utilizes
multiple
intervention
treatments
to
provide
different
barriers
for
microorganisms
to
overcome
for
survival
and
proliferation.
An
important
factor
when
using
multiple
intervention
technology
is
the
order
of
application
of
antimicrobial
treatments.
*
Corresponding
author.
Tel.:
+1-501-575-5634;
fax:
+
1-501-575-
7294.
E-mail
address:
fpohlma@comp.uark.edu
(F.W.
Pohlman).
Dorsa,
Cutter,
and
Siragusa
(1997)
reported
that
hydration
of
a
carcass
before
and
during
antimicrobial
interventions
provide
protection
to
bacteria.
Gorman,
Sofos,
Morgan,
Schmidt,
and
Smith
(1995)
reported
the
loss
of
activity
of
antimicrobial
agents
when
followed
by
plain
water
spray
washing,
possibly
due
to
physical
removal
or
dilution
of
the
sanitizing
agents.
Ellebracht
et
al.
(1999)
used
hot
water
and
lactic
acid
on
beef
trimmings
destined
for
ground
beef
to
achieved
a
reduction
of
1.1,
1.8
and
1.5
log
colony
forming
units
(CFU)/g
of
Escherichia
coli,
Salmonella
typhimurium
and
aerobic
plate
counts,
respectively.
Unfortunately,
it
remains
unknown
what
effect
other
antimicrobials
such
as
ozonated
water,
cetylpyridinium
chloride,
acetic
acid,
chlorine
dioxide
or
trisodium
phosphate,
when
used
as
combination
treatments,
might
have
on
microbial
reductions,
color
and
sensory
characteristics
of
ground
beef.
Therefore,
the
objective
of
this
research
was
to
determine
the
effectiveness
of
multiple
intervention
technologies
during
the
manufacture
of
ground
beef
on
0309-1740/02/$
-
see
front
matter
©
2002
Elsevier
Science
Ltd.
All
rights
reserved.
P11:
S0309-1740(01)00198-X
308
F.W.
Pohlman
et
al.
/Meat
Science
61
(2002)
307-313
the
reduction
of
microorganisms
and
on
instrumental
color
and
sensory
characteristics
of
ground
beef.
2.
Materials
and
methods
2.1.
Bacterial
preparation
and
inoculation
Inoculums
were
prepared
from
frozen
(-80
°C)
stock
cultures
of
E.
coli
(ATCC
11775;
EC)
and
a
nalidixic
acid
resistant
strain
of
Salmonella
typhimurium
(ATCC
1769NR;
ST).
E.
coli
was
maintained
by
brain
heart
infusion
(BHI;
Difco
Laboratories,
Detroit,
Michigan,
USA)
broth
with
glycerol
(20%)
and
Salmonella
Typhi-
murium
was
maintained
by
BHI
broth
containing
nali-
dixic
acid
(86
millimolar;
Fisher
Scientific,
Fairlawn,
New
Jersey,
USA)
with
glycerol
(20%).
Frozen
cultures
of
E.
coli
and
Salmonella
typhimurium
were
thawed,
and
0.1
ml
of
E.
coli
suspension
was
inoculated
into
separate
40
ml
aliquots
of
BHI,
and
0.1
ml
of
Salmonella
typhi-
murium
suspension
was
inoculated
into
separate
40
ml
aliquots
of
BHI
with
nalidixic
acid
(86
millimolar).
After
18
h
of
incubation
at
37
°C,
bacteria
were
har-
vested
by
centrifugation
(3649
xg
for
20
min
at
37
°C;
Beckman
GS-6
series,
Fullerton,
California,
USA),
re-suspended
in
the
same
volume
of
0.1%
buffered
peptone
water
(BPW;
Difco
Laboratories,
Detroit,
Michigan, USA)
and
then
pooled
together
(1600
ml
of
E.
coli
and
1600
ml
of
Salmonella
typhimurium)
to
make
a
bacterial
cocktail.
The
cocktail
(3200
ml;
log
10
7
colony
forming
units
(CFU)/ml
E.
coli
and
log
10
7
CFU/ml
Salmonella
typhimurium)
was
cooled
to
4
°C
and
combined
with
boneless
beef
trimmings
(12.8
kg)
and
allowed
to
attach
for
1
h
under
refrigeration
(4
°C).
The
meat
was
then
drained
and
separated
into
3.2
kg
batches
and
placed
in
a
4
°C
cooler
for
12-14
h
to
allow
further
microbial
attachment.
2.2.
Antimicrobial
treatment
application
and
sample
processing
Treatment
combinations
for
this
study
included
(1)
1%
ozonated
water
bath
(7.2
°C;
15
min)
followed
by
a
0.5%
(wt:vol)
cetylpyridinium
chloride
solution
(Zee-
land
Inc.,
Zeeland,
Michigan,
USA;
OC);
2)
1%
ozo-
nated
water
bath
(7.2
°C;
15
min.)
followed
by
a
5%
(vol:vol)
acetic
acid
solution
(Shurfine
Inc.,
Northlake,
Illinois,
USA;
OA),
(3)
200
ppm
(vol:vol)
chlorine
dioxide
solution
(Midland
Chemical
Company,
Lenexa,
Kansas,
USA),
followed
by
a
10%
(wt:vol)
trisodium
phosphate
solution
(Rhone
Poulenc,
Cranbury,
New
Jersey,
USA;
CT)
and
(4)
an
untreated
control
(C).
All
antimicrobial
treatments
were
prepared
in
deionized
water
with
the
exceptions
of
ozone
and
acetic
acid.
Ozone
was
generated
into
tap
water,
while
acetic
acid
was
commercially
prepared.
For
ozone
treatment
com-
binations,
batches
(3.2
kg)
of
inoculated
beef
trimmings
were
placed
into
a
stainless
steel
vessel
continuously
replenished
with
ozonated
water
supplied
by
an
ozone
generator
(Aqua
Air
Technologies,
Bloomfield,
New
Jersey,
USA)
for
15
min,
removed,
and
allowed
to
drip
dry
for
1
min.
The
ozonated
trimmings
were
then
placed
into
a
clean
Lyco
meat
tumbler
(Model
4Q,
Lyco
Inc.,
Janesville,
Wisconsin,
USA)
with
400
ml
of
a
selected
antimicrobial
treatment
(either
cetylpyridinium
chloride
or
acetic
acid)
and
tumbled
for
3
min
(16
rpm)
aero-
bically.
For
the
CT
treatment,
beef
trimmings
were
placed
into
a
meat
tumbler
with
400
ml
of
chlorine
dioxide,
aerobically
tumbled
for
3
min
(16
rpm),
removed
from
the
tumbler
and
placed
into
another
clean
tumbler
with
400
ml
of
trisodium
phosphate
then
tumbled
again
for
3
min
(16
rpm),
aerobically.
Upon
completion
of
the
antimicrobial
application
phase,
beef
trimmings
were
removed
from
the
tumbler
and
ground
twice
using
a
Hobart
grinder
(Model
310,
Hobart
Inc.,
Troy,
Ohio,
USA)
with
a
3.2-mm
plate.
The
ground
beef
was
divided
into
454-g
samples
and
packaged
on
stryrofoam
trays
with
absorbent
diapers.
The
trays
were
overwrapped
with
polyvinyl
chloride
film
with
an
oxygen
transmission
rate
of
1400
cc/m
2
/
2
4
h/1
atm
(Borden
Inc.,
Dallas,
Texas,
USA)
and
stored
under
simulated
retail
conditions
(4
°C;
deluxe
warm
white
fluorescent
lighting,
1630
lx,
Phillips
Inc.,
Somer-
set,
New
Jersey,
USA).
Multiple
trays
of
ground
beef
from
each
treatment
were
packaged
to
allow
for
inde-
pendent
package
use
for
microbial,
instrumental
color
and
sensory
color
and
odor
analysis
on
each
sampling
day
of
display
(days
0,
1,
2,
3,
and
7).
Fat
content
was
standardized
to
10%
and
validated
using
a
Hobart
Fat
Analyzer
(Model
F101,
Hobart
Inc.
Troy,
Ohio,
USA).
Treated
ground
beef
pH
was
also
determined
immedi-
ately
after
grinding
by
homogenizing
a
1.8-g
portion
of
ground
beef
in
18
ml
of
distilled
water
and
evaluated
using
an
Orion
Model
420A
pH
meter
with
a
ROSS
electrode
(Model
8165BN,
Orion
Research,
Inc.,
Bev-
erly,
Massachusetts,
USA).
2.3.
Microbial
sampling
On
days
0,
1,
2,
3,
and
7
of
simulated
retail
display,
25
g
of
ground
beef
was
aseptically
removed
from
the
packages
and
placed
into
whirlpack
bags
(Nasco,
Ft.
Atkinson,
Wisconsin,
USA)
with
225
ml
of
0.1%
buf-
fered
peptone
water
and
buffered
to
a
pH
of
7
with
either
sodium
hydroxide
or
hydrochloric
acid.
Samples
were
then
stomached
in
a
Model
400
Lab
Stomacher
(Seward,
London,
United
Kingdom)
for
2
min
and
serial
dilutions
were
made.
Subsequent
duplicate
plat-
ings
were
made
on
Salmonella
shigella
agar
(Difco
Laboratories,
Detroit,
Michigan,
USA)
containing
nali-
dixic
acid,
Petrifilm®
(3M
Corp.,
St.
Paul,
Minnesota,
USA)
aerobic
plate
count
(APC)
plates
and
Petrifilm®
F.W.
Pohlman
et
al.
/Meat
Science
61
(2002)
307-313
309
E.
coli/coliform
plate
count
plates.
Plates
were
then
incubated
at
37
°C
in
an
aerobic
incubation
chamber
(either
VWR
Model
5015
or
Model
3015
incubators,
VWR
Scientific,
West
Chester,
Pennsylvania,
USA)
and
APC,
Salmonella
shigella
agar
plates,
and
E.
coli
plates
were
read
at
48
h,
while
coliform
counts
were
deter-
mined
at
24
h.
Counts
were
recorded
as
colony
forming
units
per
gram.
2.4.
Instrumental
color
On
days
0,
1,
2,
3
and
7
of
simulated
retail
display,
instrumental
color
was
evaluated
using
a
HunterLab
MiniScan
XE
Spectrocolorimeter,
Model
4500L
(Hun-
ter
Associates
Laboratory
Inc.,
Reston,
West
Virginia,
USA).
Samples
were
read
using
illuminant
A/10°
obser-
ver
and
evaluated
for
CIE
(L*
a*
and
b*)
color
values.
In
addition,
reflectance
measurements
were
taken
in
the
visi-
ble
spectrum
from
580
to
630
nm.
The
reflectance
ratio
of
630
nm/580
nm
was
calculated
and
used
to
estimate
the
oxymyoglobin
proportion
of
the
myoglobin
pigment
(Hunt
et
al.,
1991;
Strange,
Benedict,
Gugger,
Metzger,
&
Swift,
1974).
In
addition,
hue
angle,
which
describes
the
hue
or
color
of
ground
beef
was
calculated
(tan'
(b*
I
a*),
as
was
the
saturation
index
((a*2
+
b*2)0.5),
which
describes
the
brightness
or
vividness
of
color
(Hunt
et
al.,
1991).
Before
use,
the
Spectrocolorimeter
was
standardized
using
white
tile,
black
tile,
and
work-
ing
standards.
Eight
measurements
were
taken
of
each
sample
and
averaged
for
statistical
analysis.
2.5.
Sensory
color
and
odor
A
six
member
trained
sensory
panel
was
used
to
evaluate
sensory
color
and
odor
characteristics
of
ground
beef
samples
through
display.
Panelists
were
selected
and
trained
by
an
experienced
panel
leader
according
to
the
American
Meat
Science
Association
guidelines
(AMSA,
1978;
Hunt
et
al.,
1991).
On
days
0,
1,
2,
3
and
7
of
simulated
retail
display,
sensory
pane-
lists
evaluated
overall
color
and
worst
point
color
(5
=
bright purplish
red,
4
=
dull
purple
red,
3
=
slightly
brownish
red,
2
=
moderately
brownish
red,
and
1=
brown)
and
percentage
surface
discoloration
(7
=
no
discoloration
(0%),
6
=
slight
discoloration
(1-20%),
5
=
small
discoloration
(20-39%),
4
=
modest
discolora-
tion
(40-59%),
3
=
moderate
discoloration
(60-79%),
2
=
extensive
discoloration
(80-95%),
1
=
total
dis-
coloration
(96-100%).
In
addition
panelists evaluated
beef
odor
(8
=
extremely
beef
like,
7
=
very
beef
like,
6
=
moderately
beef
like,
5
=
slightly
beef
like,
4
=
slightly
non-beef
like,
3
=
moderately
non-beef
like,
2
=
very
non-beef
like,
and
1
=
extremely
non-beef
like)
and
off
odor
characteristics
(5
=
no
off
odor,
4
=
slight
off
odor,
3
=
small
off
odor,
2
=
moderate
off
odor,
and
1
=
extreme
off
odor)
(Hunt
et
al.,
1991).
For
evalua-
lion,
packages
were
first
viewed
under
simulated
retail
lighting
conditions
(deluxe
warm
white
fluorescent
lighting,
1630
lx)
for
overall
color,
worst
point
color
and
percentage
discoloration.
Packages
were
then
taken
to
a
static
pressure
room,
opened,
and
evaluated
by
panelists
for
beef
odor
and
off
odor
characteristics.
2.6.
Statistical
analysis
The
experiment
was
replicated
three
times.
The
ran-
domized
complete
block
factorial
experiment
was
ana-
lyzed
using
the
GLM
procedure
of
SAS
(1988).
For
sensory
panel
data,
a
panelist
term
was
added
to
the
model
to
account
for
sensory
panelist
variation.
Treat-
ments
were
blocked
by
replicate
then
analyzed
for
the
main
effects
of
antimicrobial
treatment
combination,
day
of
display
and
appropriate
interactions.
For
vari-
ables
confounded
by
interactions,
interaction
means
were
generated,
separated
using
the
PDIFF
option
of
SAS
(1988),
and
plotted.
Least
square
means
for
all
other
variables
were
generated
and
separated
using
the
PDIFF
option
of
SAS
(1988).
3.
Results
and
discussion
3.1.
Effect
of
antimicrobial
treatment
combinations
on
microbial
populations,
instrumental
color
and
sensory
odor
characteristics
The
principle
of
multiple
intervention
technology
is
to
capitalize
on
different
weaknesses
of
various
bacterial
strains.
Placing
hurdles
such
as
pH,
chlorinated
com-
pounds
or
oxidizing
environments
in
front
of
micro-
organisms
may
cause
compromises
in
microbial
cell
wall
integrity,
metabolism
or
both,
resulting
in
a
lethal
or
inhibitory
environment
for
microbial
survival
and
pro-
liferation.
The
impact
of
multiple
interventions
on
the
reduction
of
microbial
populations
in
ground
beef
is
presented
in
Table
1.
Using
ozone
followed
by
cetyl-
pyridinium
chloride
(OC)
interventions
on
beef
trim-
mings
before
grinding
reduced
(P
<
0.05)
EC,
ST,
coliforms
(CO)
and
aerobic
bacteria
(APC)
by
1.68, 1.77,
1.88
and
1.50
log
CFU/g,
respectively
in
ground
beef
compared
with
C.
Likewise,
treatment
of
beef
trim-
mings
with
ozonated
water
followed
by
acetic
acid
(OA)
reduced
(P
<
0.05)
EC,
ST,
CO
and
APC
by
1.42, 1.66,
1.84
and
1.27
log
CFU/g,
respectively
in
ground
beef
compared
with
C.
Therefore,
it
appears
that
treatment
of
beef
trimmings
before
grinding
with
a
combination
of
a
strong
oxidant
(ozone)
and
either
a
surfactant
(cetyl-
pyridinium
chloride)
or
an
organic
acid
(acetic
acid)
were
very
effective
against
both
gram-negative
and
gram-positive
bacterium.
These
results
are
consistent
with
those
reported
by
Gorman
et
al.
(1995)
and
Graves-Delmore,
Sofos,
Schmidt,
and
Smith
(1998).
310
F.W.
Pohlman
et
al.
/Meat
Science
61
(2002)
307-313
Treatment
of
beef
trimmings
before
grinding
with
chlorine
dioxide
followed
by
trisodium
phosphate
(CT)
did
not
reduce
microorganisms
in
ground
beef
to
the
same
extent
as
the
other
combination
treatments
(Table
1).
Table
1
Effect
of
multiple
antimicrobial
treatmentsa
applied
to
beef
trimmings
on
least
square
mean
(±S.E.)
log
CFUb/g
Escherichia
coli,
coliform,
Salmonella
typhimurium,
aerobic
plate
count
(APC)
and
CIE
L*c
value,
beef
odors
and
off
odors
intensities
of
ground
beef
through
simulated
retail
display
Treatment
C
OC
OA
CT
S.E.
Microorganism
E.
coli
6.77zf
5.09w
5.35x
6.16y
0.09
Coliform
6.02z
4.14y 4.18y
5.65y
0.10
Salmonella
typhimurium
5.81z
4.04y 4.15y
5.52z
0.11
APC
7.06z
5.56x
5.79x
6.76y
0.09
Instrumental
color
CIE
L*
48.35x
52.30z
49.87y
43.80w
0.31
Sensory
odor
Beef
odor
6.44z
6.37z
3.98y
6.51z
0.14
Off
odor
4.55z
4.36z
2.54y
4.60z
0.09
a
C,
Control;
OC,
15
min
ozonated
water
bath
(1%;
7.2
°C)
and
0.5%
cetylpyridinium
chloride;
OA,
15
min
ozonated
water
bath
(1%;
7.2
°C)
and
5%
acetic
acid;
CT,
200
ppm
chlorine
dioxide
and
10%
trisodium
phosphate.
b
Colony
forming
units.
c
0
=
black
and
100
=
white.
d
Beef
odor
score:
1=
extremely
non-beef
like
and
8=
extremely
beef
like.
e
Off
odor
score:
1
=extreme
off
odor
and
5=
no
off
odor.
f
Least
square
means
within
a
row
bearing
different
letters
are
dif-
ferent
(P
<
0.05).
The
CT
treatment
reduced
(P
<
0.05)
EC,
CO
and
APC
in
ground
beef
by
0.61,
0.37
and
0.30
log
CFU/g,
respectively,
while
ST
was
not
affected
(P
>
0.05)
by
the
CT
treatment.
The
lower
bacterial
reductions
realized
by
the
CT
treatment
could
possibly
be
explained
by
the
differences
in
pH
between
the
CT,
OC,
OA
and
C
treatments.
Chlorine
dioxide's
low
pH
was
counteracted
by
trisodium
phosphate's
high
pH,
thus
resulting
in
an
overall
neutral
treatment
pH
(7.02)
for
the
CT
treat-
ment
compared
with
a
pH
of
5.72
for
C.
Therefore,
any
inhibitory
effect
due
to
individual
chlorine
dioxide
or
trisodium
phosphate
treatments
was
probably
negated
by
the
elevated
overall
pH
for
this
treatment.
The
impact
of
multiple
antimicrobial
treatments
of
beef
trimmings
before
grinding
on
instrumental
color
and
sensory
characteristics
are
shown
in
Table
1.
Ground
beef
from
the
OC
and
OA
treatments
were
lighter
(L*)
in
color
(P
<0.05),
whereas
ground
beef
from
the
CT
treatment
was
darker
(P
<0.05)
in
color
compared
with
C.
Color
differences
between
treatments
may
be
related
to
ground
beef
pH
where
both
OA
and
OC
treatments
had
a
lower
pH
(4.63
and
5.42,
respec-
tively)
than
C
(5.72)
and
the
CT
treatment
had
a
higher
pH
(7.02)
than
C.
Kaess
and
Weidemann
(1968)
found
beef
color
was
unaffected
by
low
levels
of
an
atmo-
spheric
ozone
treatment.
On
the
other
hand,
Fournaud
and
Lauret
(1972)
reported
that
atmospheric
ozone
treatment
of
beef
caused
undesirable
color
changes
to
occur.
Using
acetic
acid
as
a
single
intervention
treat-
ment,
Bell,
Marshall,
and
Anderson
(1986)
found
that
beef
cubes
discolored
immediately
upon
contact
with
this
antimicrobial.
The
OC
and
CT
treatments
were
not
different
(P>
0.05)
from
C
for
both
beef
odor
and
off
odor
Table
2
Effect
of
duration
of
display,
pooled
across
antimicrobial
treatments,
on
least
square
mean
(±SE)
log
CFUa/g
Escherichia
coli,
coliform,
Salmonella
Typhimurium,
aerobic
plate
count
(APC),
CIE
L*b
value,
beef
odorc
and
off
odors
characteristics
of
ground
beef
Days
of
display
0
1
2
3
7
Microorganism
E.
coli
5.94±0.11yze
6.08
±0.11z
5.71
±0.10y
5.80±0.10yz
5.67±0.10y
Coliform
5.22±0.12z
5.28
±0.11z
4.87±0.11y
4.82±0.11y
4.80±0.11y
Salmonella
typhimurium
5.36
±
0.13z
5.29±0.13z
4.65±0.13y
4.66±0.13y
4.44±0.13y
APC
6.20±0.10
6.29±0.10
6.24±0.10
4.44±0.13
6.51±0.10
Instrumental
color
CIE
L*
47.24
±
0.35x
47.93
±0.35xy
48.23±0.35y
49.98±0.35z 49.50±0.35z
Sensory
odor
Beef
odor
6.13±0.16yz 6.08±0.16yz
6.21±0.16z
5.78±0.16y
4.93±0.16x
Off
odor
4.21
±0.10z
4.34±0.10z
4.09±0.10yz
3.87±0.10y
3.54
±0.10x
a
Colony
forming
units.
b
0
=
black
and
100=
white.
c
Beef
odor
score:
1=
extremely
non-beef
like
and
8=
extremely
beef
like.
d
Off
odor
score:
1
=extreme
off
odor
and
5
=
no
off
odor.
e
Least
square
means
within
a
row
bearing
different
letters
are
different
(P<
0.05).
b
a
a
ab
bcb
c
:Jr
a
E#4
b=
bcb
a
_ap
a
as
=
I
A
a
d
C
abb
b
a
a
ava
aaij
a
a
a
a
a
E
A4
30
20
10
b
a
a
a
b b
b
a
bb
b
bb
a
b
a
0
0
1
2
3
7
Days
of
display
30
bb
bcc
20
=
a
a
be
aab
bb
a
b b b
a
a
i
O
P
i
10
O'
....
E
A
i
.
=
)1
1
0
C
IE
a
*
va
lue
'
(c)
a)
0
1
2
3
7
Days
of
display
630
nm
/5
80
nrn
J
4
3
2
0
(d)
a)
cv
a)
70
60
50
30
4
20
10
0
b
b
bb
b
b
b
b
a
a
a
F.W.
Pohlman
et
al.
/Meat
Science
61
(2002)
307-313
311
characteristics,
however,
ground
beef
from
the
OA
treatment
had
less
(P
<0.05)
beef
like
odor
and
more
(P
<
0.05)
off
odor
than
all
other
treatments
(Table
1).
3.2.
Effect
of
duration
of
display
on
microbial
populations,
instrumental
color
and
sensory
odor
characteristics
The
effect
of
duration
of
display,
pooled
across
anti-
microbial
treatments,
on
microbial
populations,
instru-
mental
color
and
sensory
odor
characteristics
are
summarized
in
Table
2.
By
day
7
of
display,
EC
and
APC
counts
were
similar
(P
>
0
.0
5)
to
counts
observed
on
day
1
of
display.
However,
CO
and
ST
were
reduced
(P<
0.05)
by
0.42
and
0.92
log
CFU/g,
respectively
through
7
days
of
display.
Ground
beef
became
(P
<
0.05)
lighter
(L*)
in
color,
and
had
less
(P
<0.05)
beef
odor
and
more
off
odor
from
day
1
to
day
7
of
display.
Because
EC
and
APC
remained
constant,
and
CO
and
ST
declined
through
display,
changes
in
ground
beef
color
and
odor
characteristics
may
not
be
explained
by
microbial
growth
patterns.
Instead,
pH
and
oxidative
environmental
changes
caused
by
anti-
microbial
treatments
may
have
resulted
in
myoglobin
and
lipid
oxidation,
resulting
in
color
and
odor
changes
of
ground
beef
through
display.
3.3.
Effects
of
antimicrobial
treatments
and
duration
of
display
on
instrumental
and
sensory
color
characteristics
Fig.
1
shows
the
day
of
display
by
antimicrobial
treatment
interaction
effect
on
ground
beef
redness
0
1
2
3
7
Days
of
display
40
b
bb
b
b
30
a
d
ab
b
20
10
=
0
0
1
0
1
2
3
7
Days
of
display
7
Sa
tu
ra
t
ion
in
dex
"'
2
3
Days
of
display
Ce
0
Cf
OA
9
r
:4
CTh
Fig.
1.
Day
of
display
by
antimicrobial
treatment
interaction
effect
on
the
least
square
mean
SE)
(a)
CIE
a*
value,
(b)
630
nm
reflectance/580
nm
reflectance
ratio,
(c)
CIE
b*
value,
(d)
hue
angle
and
(e)
saturation
index
of
ground
beef
through
simulated
retail
display.
abcdL
east
square
means
within
a
day
bearing
different
superscripts
are
different
(P
<
0.05).
eC,
control;
fOC,
15
min
ozonated
water
bath
(1%;
7.2
°C)
and
0.5%
cetylpyr-
idinium
chloride;
gOA,
15
min
ozonated
water
bath
(1%;
7.2
°C)
and
5%
acetic
acid;
and
h
CT,
200
ppm
chlorine
dioxide
and
10%
trisodium
phosphate.
'a*:
-60
=
green
and
+
60
=
red.
JCalculated
as
630
nm
reflectance/580
nm
reflectance.
k
b*:-60
=
blue
and
+
60
=
yellow.
'Calculated
as
tan
-1
(b*/a*).
mCalculated
as
(a*
+
b*)15.
a
C
b
111,
a
b
a
a
a
s
b
b
4
a
4
4
b%
is
b
a
A
=
A04
GOA
(a)
0
0
0
tB
a)
>
0
6
5
4
3
2
0
b
b
b b
bc
4
4
4
14
4
E
I
I
4
I
I
r4
E
Ati
a
A&
=
AM
a
a
a
C
b
a
a
0
1
2
3
7
Days
of
display
(c)
-
C
0
co
8
7
c
1
a)
0
0
1
2
3
7
Days
of
display
6
5
-0
4
3
cv
2
C
C
C
a
s
b
b
b
a
b
ab
a
a
a
4
4
1.1
14
14
4
=
V41
E
S'Oe4
A
gffit.
OA'
CT
312
F.W.
Pohlman
et
al.
/Meat
Science
61
(2002)
307-313
(a*;
panel
a)
and
oxymyoglobin
content
(630
nm/580
nm;
panel
b).
The
OC
treatment
was
less
(P
<
0.05)
red
(a*)
on
days
0
and
7
of
display
and
contained
less
(P
<
0.05)
oxymyoglobin
(630
nm/580
nm)
on
day
0
of
display,
however
was
not
different
(P
>
0.05)
on
days
1
through
3
for
either
a*
or
630
nm/580
nm
values
when
compared
with
C.
The
OA
treatment
was
initially
less
(P
<
0.05)
red
(a*)
than
C
and
lost
redness
faster
than
any
other
treatment
through
display.
Lower
redness
(a*)
values
for
the
OA
treatment
was
caused
by
a
reduced
(P
<0.05)
oxymyoglobin
(630
nm/580
nm)
content
by
day
1
of
display
for
this
treatment
compared
with
all
other
treatments.
The
lower
oxymyoglobin
content
for
the
OA
treatment
was
possibly
due
to
heme
iron
oxidation
in
the
porphyrin
ring
of
myoglobin
or
to
globin
denaturation
caused
by
the
low
pH
of
the
acetic
acid
portion
of
this
treatment.
Bell
et
al.
(1986)
also
observed
discoloration
of
beef
cubes
when
exposed
to
acetic
acid
treatment.
On
day
0
of
display,
ground
beef
from
the
CT
treat-
ment
was
less
(P
<0.05)
red
(a*;
Fig.
1,
panel
a),
how-
ever
not
different
(P
>
0.05)
in
oxymyoglobin
content
(630
nm/580
nm;
Fig.
1,
panel
b)
than
C.
Ground
beef
from
the
CT
treatment
was
similar
(P
>
0.05)
in
redness
(a*)
and
oxymyoglobin
content
(630
nm/580
nm)
on
days
1-3
of
display
to
C,
however,
by
day
7,
the
CT
treatment
remained
(P
<0
.05)
redder
(a*)
in
color
and
contained
more
(P
<0.05)
oxymyoglobin
than
all
other
treatments.
Therefore,
in
addition
to
reducing
EC,
CO
and
APC
in
ground
beef,
CT
tended
to
promote
extended
redness
and
oxymyoglobin
stability
through
display
com-
pared
to
ground
beef
made
from
untreated
trimmings
(C).
The
day
of
display
by
antimicrobial
treatment
inter-
action
effect
on
the
CIE
b*
value
is
shown
in
Fig.
1,
panel
c.
Ground
beef
from
the
OC
treatment
was
simi-
lar
(P
>
0.05)
in
yellowness
(b*)
to
C
through
display.
Likewise,
OA
was
not
different
(P
>
0.05)
in
yellowness
(b*)
on
day
0
of
display,
however
became
less
(P
<
0.05)
yellow
by
day
1
and
through
the
remainder
of
display
compared
with
C.
Ground
beef
from
the
CT
treatment
had
lower
(P
<0.05)
b*
values
on
days
0
and
7
of
dis-
play,
however,
was
not
different
(P
>
0.05)
on
days
1-3
of
display
compared
with
C.
The
hue
angle,
which
is
a
calculated
value
using
a*
and
b*
values
to
determine
the
hue
or
color
of
a
sample,
is
shown
in
Fig.
1,
panel
d.
On
day
0
of
display,
all
treatments
were
similar
(P
>
0.05)
in
hue
angle
to
C.
The
OC
and
CT
treatments
were
also
similar
(P
>
0.05)
in
hue
angle
to
C
on
days
1-3
of
display,
however,
by
day
7,
OC
had
a
larger
(P
<0.05)
hue
angle
and
CT
had
a
smaller
(P
<
0.05)
hue
angle
than
C.
The
OA
treated
samples
maintained
a
larger
(P
<0.05)
hue
angle
from
day
1—day
7
of
display
than
all
other
treatments.
The
saturation
index,
which
is
another
mathematical
calculation
using
a*
and
b*
values
to
calculate
vividness
of
color,
is
shown
in
Fig.
1,
panel
e.
Because
of
lower
a*
and
b*
values,
ground
beef
from
the
OA
treatment
was
less
(P
<0.05)
vivid
in
color
(saturation
index)
by
day
1,
and
through
the
remainder
of
display
compared
to
all
other
treatments.
Although
ground
beef
from
the
CT
treatment
was
less
(P
<
0.05)
vivid
in
color
(saturation
0
1
2
3
7
Days
of
display
4
O
3
2
1
O
Cd
oce
Fig.
2.
Day
of
display
by
antimicrobial
treatment
interaction
effect
on
the
least
square
mean
(±S.E.)
sensory
evaluated
(a)
overall
color,
(b)
worst
point
color
and
(c)
percentage
discoloration
of
ground
beef
through
simulated
retail
display.
ahcLeast
square
means
within
a
day
bearing
dif-
ferent
superscripts
are
different
(P
<
0.05).
dC,
control;
°0C,
15
min
ozo-
nated
water
bath
(1%;
7.2
°C)
and
0.5%
cetylpyridinium
chloride;
f0A,
15
min
ozonated
water
bath
(1%;
7.2
°C)
and
5%
acetic
acid;
and
gCT,
200
ppm
chlorine
dioxide
and
10%
trisodium
phosphate.
hColor
score:
1
=brown
and
5=
bright
purple
red.
Percentage
discoloration:
1=
total
discoloration
(96-100%)
and
7
=no
discoloration
(0%).
F.W.
Pohlman
et
al.
/Meat
Science
61
(2002)
307-313
313
index)
initially
(day
0),
no
difference
(P
>
0.05)
in
saturation
index
occurred
between
C,
OC
and
CT
treat-
ments
by
day
1
and
through
the
duration
of
display.
Fig.
2,
panels
a,
b
and
c
show
the
day
of
display
by
antimicrobial
treatment
interaction
effect
for
overall
color,
worst
point
color
and
percentage
discoloration,
respectively.
Sensory
panelists
found
that
ground
beef
from
the
OC
treatment
was
less
(P
<
0.05)
bright
purple
red
in
overall
color
on
days
0,
1
and
7
of
display,
how-
ever
found
no
difference
(P
>
0.05)
in
overall
color
on
days
2
and
3
of
display
compared
with
C
(panel
a).
By
day
1,
and
through
the
duration
of
display,
sensory
panelists
found
worst
point
color
to
be
similar
(P>
0.05)
between
OC
and
C
treatments
(panel
b).
Likewise,
by
day
1
of
display,
no
difference
(P>
0.05)
in
percentage
discoloration
was
evident
between
OC
and
C
treatments
until
day
7
of
display
(panel
c).
In
contrast,
sensory
panelists
indicated
that
ground
beef
from
the
OA
treat-
ment
was
less
(P<
0.05)
bright
purple
red
in
overall
(panel
a)
and
worst
point
(panel
b)
color,
and
had
a
larger
(P
<
0.05)
percentage
surface
discoloration
(panel
c)
than
C
throughout
display.
Beef
treated
with
CT
was
not
different
(P
>
0.05)
from
C
on
days
0-2
of
display
for
overall
color
(Fig.
2,
panel
a)
and
percentage
discoloration
(Fig.
2,
panel
c).
How-
ever,
the
CT
treatment
was
brighter
purple
red
(P
<
0.05)
in
overall
color
(Fig.
2,
panel
a)
and
had
less
(P
<
0.05)
percentage
discoloration
(Fig.
2,
panel
c)
than
C
on
days
3
and
7
of
display.
In
addition,
the
CT
samples
maintained
a
brighter
(P
<0.05)
purple
red
worst
point
color
than
all
other
treatments
throughout
display
(Fig.
2,
panel
b).
Therefore,
treatment
of
beef
trimmings
before
grinding
with
chlorine
dioxide
and
CT
not
only
reduced
microorganisms
in
ground
beef,
but
also
improved
color
stability
and
shelf
life
extension.
4.
Conclusion
Multiple
antimicrobial
intervention
treatment
combi-
nations
utilizing
ozone,
chlorine
dioxide,
cetylpyr-
idinium
chloride
and
trisodium
phosphate
on
beef
trimmings
before
grinding
can
be
used
to
effectively
reduce
bacterial
numbers
with
little
effect
on
fresh
ground
beef
color
and
odor
characteristics.
Therefore,
the
inclusion
of
multiple
antimicrobial
intervention
technologies
into
ground
beef
processing
systems
could,
if
approved,
provide
an
added
measure
of
safety
to
ground
beef
without
negatively
impacting
fresh
product
sensory
characteristics.
Acknowledgements
Appreciation
is
expressed
to
the
Arkansas
Beef
Council
for
funding
this
research.
The
authors
would
like
to
thank
J.
Davis,
L.
Rakes,
A.
Ivey,
L.
McBeth,
R.
Story
and
E.
Kroger
for
their
assistance
in
conducting
these
trials.
References
American
Meat
Association
(AMSA).
(1978).
Guidelines
for
cookery
and
sensory
evaluation
of
meat.
Chicago,
IL:
Am.
Meat
Sci.
Assoc.
and
National
Live
Stock
and
Meat
Board.
Bell,
M.
F.,
Marshall,
R.
T.,
&
Anderson,
M.
E.
(1986).
Micro-
biological
and
sensory
tests
of
beef
treated
with
acetic
and
formic
acids.
Journal
of
Food
Protection,
49(3),
207-210.
Dorsa,
W.
J.,
Cutter,
C.
N.,
&
Siragusa,
G.
R.
(1997).
Effects
of
acetic
acid,
lactic
acid
and
trisodium
phosphate
on
the
microflora
of
refrigerated
beef
carcass
surface
tissue
inoculated
with
Escherichia
coli
0157:H7,
Listeria
innocua,
and
Clostridium
sporogenes.
Journal
of
Food
Protection,
60(6),
610-624.
Ellebracht,
E.
A.,
Castillo,
A.,
Lucia,
L.
M.,
Miller,
R.
K.,
&
Acuff,
R.
G.
(1999).
Reduction
of
pathogens
using
hot
water
and
lactic
acid
on
beef
trimmings.
Journal
of
Food
Science,
64(6),
1094-1099.
Foumaud,
J.,
&
Lauret,
R.
(1972).
Influence
of
ozone
on
the
surface
microbial
flora
of
beef
during
refrigeration
and
thawing.
Tecnologia-
Alimentaria,
6(35/36),
12-14,16.
Gorman,
B.
M.,
Sofos,
J.
N.,
Morgan,
J.
B.,
Schmidt,
G.
R.,
&
Smith,
G.
C.
(1995).
Evaluation
of
hand-trimming,
various
sanitizing
agents,
and
hot
water
spray-washing
as
decontamination
interven-
tions
for
beef
brisket
adipose
tissue.
Journal
of
Food
Protection,
58(8),
899-907.
Graves-Delmore,
L.
R.,
Sofos,
J.
N.,
Schmidt,
G.
R.,
&
Smith,
G.
C.
(1998).
Decontamination
of
inoculated
beef
with
sequential
spray-
ing
treatments.
Journal
of
Food
Science,
63(5),
890-893.
Hunt,
M.
C.,
Acton,
J.
C.,
Benedict,
R.
C.,
Calkins,
C.
R.,
Corn-
forth,
D.
P.,
Jeremiah,
L.
E.,
Olson,
D.
G.,
Salm,
C.
P.,
Savell,
J.
W.,
&
Shivas,
S.
D.
(1991).
AMSA
guidelines
for
meat
color
evaluation.
In
E.
Schofield
(Ed.),
Proceedings
44th
Annual
Reciprocal
Meat
Conference,
9-12
July
1991.
Kansas
State
University,
Man-
hattan,
Kansas,
Chicago,
IL:
National
Live
Stock
and
Meat
Board
(pp.
3-17).
Kaess,
G.,
&
Weidemann,
J.
F.
(1968).
Ozone
treatment
of
chilled
beef.
I.
Effect
of
low
concentrations
of
ozone
on
microbial
spoilage
and
surface
colour
of
beef.
Journal
of
Food
Technology,
3(4),
325-
334.
Kim,
J.
W.,
&
Slavik,
M.
F.
(1994).
Trisodium
phosphate
(TSP)
treatment
of
beef
surfaces
to
reduce
Escherichia
coli
0157:H7
and
Salmonella
typhimurium.
Journal
of
Food
Science,
59(1),
20-22.
Kochevar,
S.
L.,
Sofos,
J.
N.,
LeValley,
S.
B.,
&
Smith,
G.
C.
(1997).
Effect
of
water
temperature,
pressure
and
chemical
solution
of
removal
of
fecal
material
and
bacteria
from
lamb
adipose
tissue
by
spray-washing.
Meat
Science,
45(3),
377-388.
Phebus,
R.
K.,
Nutsch,
A.
L.,
Schaffer,
D.
E.,
Wilson,
C.
R.,
Reim-
ann,
M.
J.,
Leising,
J.
D.,
Kastner,
C.
L.,
Wolfe,
J.
R.,
&
Prasai,
R.
K.
(1997).
Comparison
of
steam
pasteurization
and
other
meth-
ods
for
reduction
of
pathogens
on
surfaces
of
freshly
slaughtered
beef.
Journal
of
Food
Protection,
60(5),
476-484.
Reagan,
J.
0.,
Acuff,
G.
R.,
Buege,
D.
R.,
Buyck,
M.
R.,
Dickson,
J.
S.,
Kastner,
C.
L.,
Marsden,
J.
L.,
Morgan,
J.
B.,
Nickelson
II,
R.,
Smith,
G.
C.,
&
Sofos,
J.
N.
(1996)
Trimming
and
washing
of
beef
carcasses
as
a
method
of
improving
the
microbiological
quality
of
meat.
Journal
of
Food
Protection,
59(7),
751-756.
SAS.
(1988).
SAS/STAT
user's
guide:
release
6.12.
Cary,
NC:
SAS
Institute,
Inc.
Strange,
E.
D.,
Benedict,
R.
C.,
Gugger,
R.
E.,
Metzger,
V.
G.,
&
Swift,
C.
E.
(1974).
Simplified
methodology
for
measuring
meat
color.
Journal
of
Food
Science,
39,
988-992.