ARTICLE IN PRESS

FOOD
MICROBIOLOGY
Food Microbiology 23 (2006) 423–429
www.elsevier.com/locate/fm

Shelf-life of a chilled precooked chicken product stored in air and under

modified atmospheres: microbiological, chemical, sensory attributes
A. Patsias, I. Chouliara, A. Badeka, I.N. Savvaidis, M.G. Kontominas
Laboratory of Food Chemistry and Food Microbiology, Section of Industrial and Food Chemistry, Department of Chemistry,
University of Ioannina, Ioannina 45110, Greece
Received 29 March 2005; received in revised form 25 July 2005; accepted 4 August 2005
Available online 18 October 2005

Abstract

This study evaluated the effect of modified atmosphere packaging on shelf-life extension of a precooked chicken meat product stored
at 41C using microbiological, physico-chemical and sensory analyses. The following gas mixtures were used: M1: 30%/70% (CO2/N2),
M2: 60%/40% (CO2/N2) and M3: 90%/10% (CO2/N2). Identical chicken samples were aerobically packaged and used as control
samples. Sampling was carried out at predetermined time intervals namely: 0, 4, 8, 12, 16 and 20 days. Total viable counts (TVC), Lactic
acid bacteria (LAB), Brochothrix thermosphacta, pseudomonads, yeasts and molds, and Enterobacteriaceae were monitored. TVC of
precooked chicken product reached 7 log cfu/g, after days 12 and 16 of storage (air and M1 samples), respectively. The M2 and M3 gas
mixture packaged samples did not reach this value throughout the 20 days storage period under refrigeration. LAB and to a lesser degree
B. thermosphacta, constituted part of the natural microflora of precooked chicken samples stored in air and under MAP reaching
7.0–8.1 log cfu/g at the end of storage period. Of the remaining bacterial species monitored, both pseudomonads and yeasts/molds were
significantly higher ðPo0:05Þ for chicken samples stored in air than under MAP (M1, M2, M3) throughout the entire storage period
under refrigeration. Finally, counts of Enterobacteriaceae were low (o2 log cfu/g) in all chicken samples irrespective of the packaging
conditions throughout the entire storage period. Of the chemical indices determined, thiobarbituric (TBA) values in all cases remained
low, equal or lower than 3.0 mg malonaldehyde (MA)/kg during the entire storage period. Results of the present work show that the limit
of sensory acceptability was only reached for the aerobically stored and M1 gas mixture chicken samples somewhat before days 16 and 20
of storage, respectively. This limit coincided with high TVC and LAB populations (46.8 log cfu/g), increased lipid oxidation (aerobic
storage only) and apparent growth of yeasts/moulds on the surface of chicken samples. The use of MAP as shown in the present study,
resulted in an extension of shelf-life of precooked chicken by ca. 4 days (M1 gas mixture), and by more than 6 days (M2 and M3 gas
mixtures), respectively. Precooked chicken meat was better preserved under M2 and M3 mixtures maintaining desirable odor/taste
attributes even on final day of storage tested.
r 2005 Elsevier Ltd. All rights reserved.

Keywords: Precooked Chicken; Shelf-life; Packaging; Spoilage

1. Introduction available as either fresh or precooked (i.e. fried) chicken

Poultry products are highly perishable foods. Depending
on the degree of processing following slaughter, their
spoilage varies between 4 and 10 days under refrigeration
(Marenzi, 1986). In the last decade, chicken-based meat
products have become increasingly popular worldwide due
to their high nutritional quality and low cost and are
Corresponding author. Tel.: +30 265 10 98342; fax: +30 265 10 98795.
E-mail address: mkontomi@cc.uoi.gr (M.G. Kontominas).
and/or microbiological products, which after subsequent
packaging are usually stored under refrigeration (Barbut,
2002). Additionally, frozen chicken-based meat products
also available on the market include specialties such as:
nuggets, meatballs, hamburgers, frankfurters, etc.
Susceptibility of chicken meat and chicken-based meat
products to microbial spoilage presents a potential health
hazard, since poultry meat may harbor pathogenic micro-
organisms (Geornaras et al., 1998). Spoilage is commonly
detected by sensory and/or micro-biological analysis

0740-0020/$ - see front matter r 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.fm.2005.08.004
 ARTICLE IN PRESS
424 A. Patsias et al. / Food Microbiology 23 (2006) 423–429
(Dainty, 1996). A disadvantage of the former often requires
trained panelists to minimize subjectivity, whereas the
latter is laborious, time consuming and requires extensive
knowledge of the specific spoilage organisms. An alter-
native method to the above-mentioned analyses involves
the measurement of chemical changes associated with the
growth of specific spoilage organisms in meat and meat
products (Dainty, 1996). Among these changes, lactate,
glucose, degree of lipid oxidation, and biogenic amines
(BAs) have been proposed as potential indicators of
spoilage (de Azevedo et al., 2003; Patsias et al., 2005).
Modified atmosphere packaging (MAP) has gained
considerable popularity over the last decades as a modern
non-thermal method of food preservation. The proper
combination of gases (CO2, N2 and O2) in the headspace of
food packs results in suppression of the microbial flora of
perishable foods such as meat, fish and related products,
developed under aerobic conditions and retention of their
sensorial attributes (Davies, 1995). A minimum CO2
concentration of 20 – 30% is necessary to exhibit an
inhibitory effects (Stiles, 1991). An extended storage-life of
poultry meat under low O2, high CO2 concentrations is
observed because the spoilage caused by LAB occurs later
as compared to the spoilage caused by aerobic bacteria
such as Pseudomonas spp. that prevail under aerobic
conditions. In general, the defect caused by LAB is
described as ‘souring’, which is less offensive than the
putrefaction that develops aerobically (Stiles, 1991).
Microflora and spoilage pattern of chicken carcasses
packaged under CO2 conditions are similar to those
observed for red meat (Gill, 1986).
Despite the work on preservation of fresh poultry meat
using (MAP), (Sawaya et al., 1995; Kim and Marshall,
1999; Saucier et al., 2000) alone or in combination with
other methods including: treatment with acids (Jimenez et
al., 1999), EDTA-nisin treatment (Cosby et al., 1999),
addition of phosphates (Hwang and Beauchat, 1995),
essential oils (Carlos and Harrison, 1999), high hydrostatic
pressure treatment (Linton et al., 2004) and irradiation
(Mahrour et al., 2003; Nam and Ahn, 2003), limited data
are available in the literature on the effect of MAP on
spoilage microflora of precooked chicken meat products.
In contrast, there are several studies focusing on the fate/
survival of pathogens in MA packaged precooked chicken
meat products (Marshall et al., 1991, 1992, Wel et al.,
2001). Thus, the objective of the present work was to study
the effect of MAP including air on the shelf-life and quality
of a precooked chicken meat product stored at 4 1C using
microbiological, physico-chemical and sensory analyses.
2. Materials and methods
2.1. Preparation of precooked chicken samples and storage
conditions
Fresh chicken breast fillets were prepared by massaging
and forming by a local poultry processing plant (Pindos
SA, Ioannina, Greece) ca. 8 h after slaughter. Breast fillets,
(portions of 100 g) were dipped in a batter, coated with
rusk granules and fried at 180 1C for 2 min in a CFC, model
GR 3000/40 fryer (CFS, Utrecht, The Netherlands) using
corn/sunflower (1:1 v/v) frying oil. Samples were then
aseptically transferred to a CFS, model HLT 8000/600
oven (CFS, Utrecht, The Netherlands) and baked (air
temperature 150 1C, 5 min) until the core of the product
reached an internal temperature of at least 75 1C and
immediately cooled at
1 1C according to the company
recipe using standard industrial procedures. Precooked
(‘Schnitzel’ type) products were transported to the labora-
tory in insulated polystyrene boxes on ice within 1 h of the
baking process. The precooked chicken samples (ca. 300 g)
were then placed in low-density polyethylene/polyamide/
low-density polyethylene (LDPE/PA/LDPE) barrier
pouches (1 fillet/pouch), 75 mm in thickness having an
oxygen permeability of 52.2 ml/m2 day atm at 60% RH/
251C and water vapour permeability of 2.4 g/m2 day at
100% RH/251C. The following gas mixtures were used:
M1: 30%/70% (CO2/N2), M2: 60%/40% (CO2/N2) and
M3: 90%/10% (CO2/N2). Gas mixtures were prepared
using a PBI-Dansensor model 9000 gas mixer (Ringsted,
Denmark). Pouches were heat-sealed using a BOSS model
N48 vacuum sealer (Boss GmbH, Germany) and kept
under refrigeration ð4  0:5 o CÞ. Gas concentrations in
packages were determined using a BPI, model checkmate
9900 headspace analyzer instrument (Ringsted, Denmark).
Identical chicken samples were aerobically packaged and
used as control samples. Control samples were packaged in
a polyethylene film, 75 mm in thickness, having an oxygen
permeability of 4600 cm3/(m2 day atm) at 60% RH/25 1C
and a water vapor permeability of 1.75 g/(m2 day atm) at
100% RH/25 1C. Sampling was carried out at predeter-
mined time intervals namely: 0, 4, 8, 12, 16 and 20 days
based on information provided by the company quality
control laboratory according to which the product shelf life
packaged in air was 12–13 days.
2.2. Microbiological analysis
A sample (25 g) was removed aseptically using a scalpel
and forceps from the breaded precooked chicken breast
fillet, transferred to a stomacher bag (Seward Medical,
UK), containing 225 ml of sterile quarter-strength Ringer’s
solution, and homogenized using a stomacher (Lab
Blender 400, Seward Medical, UK) for 60 s at room
temperature. For microbial enumeration, 0.1 ml samples of
serial dilutions (1:10, diluent, quarter-strength Ringer’s
solution) of chicken homogenates were spread on the
surface of dry media. Total viable counts (TVC) were
determined using Plate Count Agar (PCA, Merck, Den-
mark, Germany), after incubation for 3 days at 30 1C.
Pseudomonads were determined on cetrimide fusidin
cephaloridine agar (Oxoid supplemented with selective
supplement SR 103, Oxoid, Basingstoke, UK) after
incubation at 25 1C for 2 days (Mead and Adams, 1977).
 ARTICLE IN PRESS
A. Patsias et al. / Food Microbiology 23 (2006) 423–429
Brochothrix thermosphacta was determined on streptomy-
cin sulphate–thallous acetate–cycloheximide (actidione)
agar, prepared from basic ingredients in the laboratory
after incubation at 25 1C for 3 days (Gardner, 1966). For
members of the Enterobacteriaceae family, 1.0 ml sample
was inoculated into 10 ml of molten (45 1C) violet red bile
glucose agar (Oxoid). After setting, a 10 ml overlay of
molten medium was added and incubated at 30 1C for 24 h.
The large colonies with purple haloes were counted (Mossel
et al., 1979). Lactic acid bacteria (LAB) were determined
on de Man Rogosa Sharpe medium (Oxoid) after incuba-
tion at 25 1C for 5 days. Yeasts and molds were enumerated
using rose bengal chloroamphenicol agar (RBC, Merck)
after incubation at 25 1C for 3 days in the dark. All plates
were examined visually for typical colony types and
morphological characteristics associated with each growth
medium. In addition, the selectivity of each medium was
checked routinely by Gram staining and microscopic
examination of smears prepared from randomly selected
colonies from all of the media.
2.3. Chemical analysis
The pH value was recorded using a Metrohm, model
691, pH meter. Chicken samples were thoroughly homo-
genized with 10 ml of distilled water and the homogenate
used for pH determination. TBA was determined according
to the method proposed by Pearson (1976). Thiobarbituric
TBA content was expressed as milligrams of malonalde-
hyde (MA)/ kg chicken.
2.4. Sensory evaluation
After each individual sampling, samples were frozen
until sensory evaluation. Each precooked chicken breast
fillet (ca. 100 g) was reheated in a microwave oven at high
power (700 W) for 4 min including time of defrosting. A
panel of seven judges experienced in chicken evaluation
was used for sensory analysis. Panelists were trained for a
period of 3 months in 1-h sessions three times a week (36 h
total) (Winger and Pope, 1976). Triangle tests were
performed in order to select seven panelists who could
detect off-flavors in precooked chicken. Prior to sample
evaluation, the seven selected panelists participated in
orientation sessions to familiarize with the flavor attributes
(off-odor, off-taste) of precooked chicken. Panelists were
asked to evaluate taste and odor intensities of the heated
samples. Along with the test samples, the panelists were
presented with a freshly thawed chicken sample, stored at

30 1C throughout the experiment, this serving as the
reference sample. Acceptability as a composite of odor and
taste was estimated using a descriptive scale ranging from
1–9, where: 1 ¼ extreme foreign flavor or dislike intensely
and 9 ¼ no foreign flavor or like extremely. A score of 6
was taken as the lower limit of acceptability (Boerema et
al., 1993; Penney et al., 1993). The product was defined as
425
unacceptable after development of first off-odor or off-
taste as scored by at least 50% of the judges.
2.5. Statistical analysis
Experiments were replicated twice on different occasions
with different precooked chicken samples. Different
packages were sampled on predetermined time intervals.
Analyses were run in triplicate for each replicate
ðn ¼ 2  3Þ. Microbiological data were transformed into
logarithms of the number of colony forming units (cfu/g)
and were subjected to analysis of variance (ANOVA).
Means and standard deviations were calculated, and, when
F-values were significant at the Po0:05 level, mean
differences were separated by the Least Significant
Difference (LSD) procedure (Steel and Torrie, 1980).
3. Results
3.1. Microbiological changes
The present study focused on the monitoring of the
following species of micro-organisms: TVC, LAB, B.
thermosphacta, pseudomonads, Enterobacteriaceae, and
yeasts and molds. TVC of precooked chicken product
(Fig. 1a) was ca. 3.9 log cfu/g (day 0). Initial TVC reached
the value of 7 log cfu/g, which is considered as the upper
acceptability limit for fresh poultry meat as defined by
ICMSF (1986) ca. on days 12 and 16 of storage (air and
M1 samples), respectively. The M2 and M3 gas mixture
packaged samples did not reach this value throughout the
20 days storage period under refrigeration. After 20 days of
storage, the M3 gas mixture contributed to significantly
lower ðPo0:05Þ TVC count than the M1 and air samples.
Of the facultative anaerobic bacterial species, LAB and
to a lesser degree B. thermosphacta constituted part of the
natural microflora of precooked chicken samples stored in
air and under MAP (M1) (Fig. 1b and c), respectively. Of
these bacterial species, LAB was a dominant bacterial
species and constituted a major part of the microbial
association of precooked chicken product irrespective of
the packaging conditions (Fig. 1b). Initial (day 0) LAB
counts determined were low, 2.7 log cfu/g, increasing
progressively with storage time attaining final counts of
ca. 7.5–8.1 log cfu/g for M1, M2 and M3 gas mixture
packaged samples. Air-packaged samples attained final
values of 7 log cfu/g. Interestingly, a different trend was
noted for B. thermosphacta for which much higher counts
ðPo0:05Þ were recorded in air-packaged chicken samples
as compared to the M1 gas mixture samples throughout the
entire storage period. Respective counts of M2 and M3 gas
mixture packaged samples remained lower than 2.0 log cfu/
g throughout the 20 day storage period.
Of the remaining bacterial species determined, both
pseudomonads and yeasts/molds although strictly aerobic,
were also found in the microbial association of the pre-
cooked chicken product; however, significantly higher counts
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426 A. Patsias et al. / Food Microbiology 23 (2006) 423–429

9 8
8 7
7 6
6
log cfu /g

log cfu /g
5
5
4
4
3 3
2 2
1 1
0 0
0 4 8 12 16 20 24 0 4 8 12 16 20 24
(a) Storage time (days) (c) Storage time (days)

9 6
8
5
7
6
log cfu /g

log cfu /g
5
3
4
3 2
2
1
1
0 0
0 4 8 12 16 20 24 0 4 8 12 16 20 24
(b) Storage time (days) (d) Storage time (days)

7
6
5
log cfu /g

4
3
2
1
0
0 4 8 12 16 20 24
(e) Storage time (days)

Fig. 1. Changes (log cfu/g) in total viable count (a); lactic acid bacteria (b); Brochothrix thermosphacta (c); Pseudomonas spp. (d) and yeasts and molds (e)
of chilled precooked chicken meat packaged in air (’) and under MAP. Gas mixtures: M1, (~); M2, (m) and M3 (K). Each point is the mean 7 SE
(0.2–0.4) of two replicate experiments with three samples analyzed per replicate ðn ¼ 6Þ.

ðPo0:05Þ were recorded for chicken samples stored in air
than under M1, M2 and M3 samples throughout the entire
storage period under refrigeration (Fig. 1d and e). Initial (day
0) counts for both these species were low (o 2 log cfu/g) and
increased progressively with storage time for air-packaged
chicken samples attaining final counts of 5.5–6.2 log cfu/g on
final day 20 of storage. For Pseudomonas spp., counts of M1,
M2 and M3 gas mixture samples remained low (o 2 log cfu/
g) throughout the 20 day storage period (Fig. 1d). Yeasts/
molds, were inhibited by MAP attaining values of ca.
2.3–3.2 log cfu/g on final day 20 of storage (Fig. 1e). The
higher the CO2 concentration in the MAP gas mixture, the
higher the inhibition recorded. Finally, counts of Enterobac-
teriaceae were also low (o 2 log cfu/g) in all chicken samples
irrespective of the packaging conditions throughout the entire
storage period (results not shown).
3.2. Chemical changes
Changes in pH during storage of chicken samples both in
air and under MAP conditions were not statistically
significant ðP40:05Þ. Values of pH for air and M1, M2
and M3 gas mixture packaged chicken samples were in the
range of ca. 6.25–6.42 (results not shown).
TBA values of precooked chicken samples are presented
in Fig. 2. Interestingly, TBA values of air and MA-
packaged chicken samples decreased up to day 8 of
refrigerated storage. Beyond this point in time TBA values
significantly ðPo0:05Þ increased for air-packaged chicken
samples while those obtained for M1, M2 and M3 gas
mixture packaged samples remained more or less constant
(Fig. 2). TBA values in all cases remained lower or equal to
3.0 mg malonaldehyde (MA)/kg during the entire storage
period of 20 days (Fig. 2).
3.3. Sensory analysis
The results of the sensory evaluation (odor and taste
attributes) of the microwave heated chicken product are
presented as overall acceptability scores (Fig. 3). Combined
scores for odor and taste showed a similar pattern of
decreasing acceptability (individual results not shown).
 ARTICLE IN PRESS
TBA (mg malonaldehyde / kg) A. Patsias et al. / Food Microbiology 23 (2006) 423–429 427

4 processed chicken products. Of the three gas mixtures used

in the present study, M2: 60%/40% and M3: 90%/10%
3 (CO2/N2) gases were the most effective for controlling
TVC. For these gas mixtures, TVC counts remained under
2 the maximum value of 7 log cfu/g during the entire storage
period of 20 days. It is now well established that
1 atmospheres containing CO2 either alone or in combina-
tion with nitrogen delay the growth of typically aerobic
0
0 4 8 12 16 20 24 spoilage microflora by inhibiting the growth of aerobic
Storage time (days) Gram-negative bacteria such as Pseudomomas spp. and
also yeasts/molds. This is a result of an extension of lag
Fig. 2. Changes in thiobarbituric acid (TBA) (mg of malonaldehyde/kg)
phase of growth, and a decrease in the growth rate during
of chilled precooked chicken meat packaged in air (’) and under MAP.
Gas mixtures: M1, (~); M2, (m) and M3 (K). Each point is the mean 7 the logarithmic phase (Farber, 1991). In the present study,
SE (0.05–0.44) of two replicate experiments with three samples analyzed CO2 atmospheres and aerobic storage as expected, favored
per replicate ðn ¼ 6Þ. the dominance of facultative anaerobic population i.e.
LAB and of aerobic Pseudomonads in precooked chicken
samples, respectively. Present results indicate that LAB
10 were an important part of the precooked chicken spoilage
microflora, irrespective of the packaging conditions prob-
8
Sensory score

ably originating either from post-heat treatment cross-

6 contamination or partial survival of certain thermotolerant
4
2 chicken samples stored under aerobic conditions. The
0
0 4 8 12 16
Storage time (days)
Fig. 3. Changes in overall acceptability scores (mean values of taste and
odor) of chilled precooked chicken meat packaged in air (’) and under
MAP. Gas mixtures: M1, (~); M2, (m) and M3 (K). Each point is the
mean 7 SE (0.1–0.3) of two replicate experiments with three samples
analyzed per replicate ðn ¼ 6Þ.
Air-packaged chicken samples received higher overall
acceptability scores ðPo0:05Þ than M1, M2 and M3 gas
packaged samples up to day 12, whereas after this time
period this trend was reversed continuing throughout the
entire period of refrigerated storage. All chicken samples
received high scores during the first 12 days, while after this
period significant differences ðPo0:05Þ were observed in
sensory scores between air and MA-packaged samples. The
limit of overall acceptability (score 6) was reached some-
what before day 16 (air samples) and day 20 (M1 samples),
while M2 and M3 samples never reached this limit within
the time span of the experiment (Fig. 3).
4. Discussion
In the present study initial (3.9 log cfu/g) populations of
aerobic micro-organisms (TVC) for precooked chicken
indicate probable post-cooking cross-contamination as
well as a high initial (prethermal treatment) microbial load
(105–106 cfu/g) given that during the pasteurization treat-
ment only a 2 log reduction in TVC is expected. Despite
this fact the product is still considered acceptable in terms
of TVC. Similarly, Ismail et al. (2000) reported mean TVC
populations of 3.32–5.77 log cfu/g for various raw and
species after thermal processing (Holzapfel, 1998), whereas
the contribution of pseudomonads was significant only for
suppression of aerobic bacteria i.e. pseudomonads using
20 24 MAP can be beneficial in the sense that the end-products of
LAB and/or B. thermosphacta are relatively less offensive
as compared to the typical spoilage odors produced by
Pseudomonas spp. Surprisingly, B. thermosphacta was only
detected in chicken stored under aerobic conditions and to
a lesser extent in chicken meat stored under CO2 (mixture
M1). This bacterium is known to grow under both aerobic
and anaerobic conditions, and has been implicated in raw
chicken meat spoilage (Kakouri and Nychas, 1994)
producing cheesy or dairy odors (Dainty and Mackey,
1992). It is likely that increasing populations of this
bacterium in air packaged and M1 samples may be due
to probable cross-contamination after thermal processing
treatment of chicken samples, whereas suppression of B.
thermosphacta growth in precooked chicken samples
packaged under M2 and M3 gas mixtures, is due to high
numbers (5–8 log cfu/g) of LAB between days 8 and 20 of
storage, as also reported for vacuum and MA-packaged
beef (Nissen et al., 1996). The presence of Brochothrix spp.
in large populations is highly undesirable because of its
high spoilage potential (Gill, 1986). It must be stated that
there is only limited information concerning the implica-
tion of this bacterium in the spoilage of meat, including
poultry and products stored under aerobic and CO2
conditions. Of the microbial associations developed in
chicken meat under aerobic and CO2 storage, yeasts/molds
were found to be a member, and along with LAB, B.
thermosphacta, and to a lesser extent Pseudomonads, play a
significant role in spoilage of precooked chicken. Accord-
ing to Roth and Clark (1976) the extension of product
shelf-life afforded by the use of carbon dioxide may be
attributed largely to the effective control of the growth of
 ARTICLE IN PRESS
428 A. Patsias et al. / Food Microbiology 23 (2006) 423–429
Brochothrix spp. Interestingly in our study growth of
yeasts/fungi was observed in precooked chicken samples
stored aerobically between days 16 and 20 of refrigerated
storage corresponding to yeasts/mold populations of ca.
6 log cfu/g. Present results obtained for yeasts/molds in
chicken meat are in agreement with results obtained by
Ismail et al. (2000) for various chicken products. It is
noteworthy that, populations of yeasts and specific yeast
species in fresh and processed poultry products have been
determined only in a few studies and so far limited
information is available on identification of yeasts and
moulds and their association to poultry spoilage (Deak and
Beuchat, 1996). Knowledge of the presence and numbers of
yeasts and moulds in poultry and processed poultry
products would be useful when developing technologies
to retard spoilage of poultry. Interestingly, in the present
study, low numbers of Enterobacteriaceae (o 2.0 log cfu/g)
in precooked chicken, noted throughout the entire storage
period, irrespective of packaging conditions (aerobic and
modified atmosphere), may be due to the inherent
sensitivity of Gram-negative bacteria, including this genus
to extrinsic factors such as heat, aw, pH, etc. (Holzapfel,
1998) (results not shown). One increasingly important
aspect of extended storage of chilled precooked products is
microbiological safety (Farber, 1991). Packaging atmo-
spheres rich in carbon dioxide have been shown to slow
growth of mesophilic pathogens in products such as sliced
roast beef stored at temperatures above 10 1C (Hintlian and
Hotchiss, 1987). On the other hand, cold tolerant patho-
gens such as Listeria monocytogenes can under certain
circumstances outgrow spoilage bacteria on raw and
cooked chicken products (Ingham et al., 1990, Marshall
et al., 1991) stored under MAP. Since spoilage may follow
pathogen growth the microbiological safety of MAP foods
continues to be questioned (Farber, 1991).
Gill and Reichel (1989) found that cold tolerant
pathogens such as Listeria monocytogenes, Yershinia
enterocolitica and Aeromonas hydrophila either did not
grow or grew more slowly on high pH raw meat in oxygen-
free carbon dioxide packaging atmospheres than on
vacuum packed meat. Therefore the potential for growth
of cold tolerant pathogens on cooked meat remains to be
determined and safe storage temperature regimes need to
be established before the use of CO2 packs can be safely
recommended for the extended chilled storage of cooked
meats.
TBA is a measure of malonaldehyde (MA), one of the
degradation products of lipid hydroperoxides formed
through oxidation of unsaturated fatty acids (UFA)
(Nawar, 1996). As can be seen in Fig. 2 there is a trend
towards a decrease in TBA values up to day 8, irrespective
of packaging conditions, followed by an increase after day
8 of storage for both aerobically and modified atmosphere
packaged chicken samples. The reason for this trend is
unclear. It is interesting to note that the onset of increase in
TBA values of aerobically packaged chicken after day 16 of
storage coincided with the dramatic decrease in the overall
acceptability score reaching its lower limit (Fig. 3).
Presence of oxygen is the most critical factor influencing
lipid oxidation. Present results show that aerobic storage
enhances lipid oxidation, whereas MAP has the potential
to control lipid oxidation in chicken. In related studies,
increasing CO2 concentrations (30%, 70%) were effective
in inhibiting the production of free fatty acids in fresh
poultry meat (Sawaya et al., 1995).
Results of the present work show that the limit of
sensory acceptability (a score of 6), was only reached for
the aerobically stored and M1 gas mixture chicken samples
somewhat before days 16 and 20 of storage, respectively.
This limit coincided with high TVC and LAB populations
(46.8 log cfu/g) (Fig. 1a and b), increased lipid oxidation
(aerobic storage only) and apparent growth of yeasts/
moulds on the chicken samples. The use of CO2 atmo-
spheres as shown in the present study, resulted in an
extension of shelf-life of precooked chicken by ca. 4 days
(M1 gas mixture), and by more than 6 days (M2 and M3
gas mixtures), respectively. Overall acceptability data (Fig.
3) of air and MA-packaged chicken samples correlated
rather well with TVC data (Fig. 1a). Precooked chicken
meat was better preserved under M2 and M3 mixtures
maintaining acceptable odor/taste attributes even on final
day of storage.
5. Conclusions
Although the combination of precooking treatment and
subsequent aerobic storage of chicken gives a good-quality
poultry product with acceptable shelf-life (14–15 days), as
judged by both and sensory and microbiological analyses,
the use of MAP combined with precooking treatment may
further increase the shelf-life of the precooked chicken
product by at least 6 days.
Acknowledgements
The authors would like to thank PINDOS S.A.,
Ioannina, Greece for providing chicken samples.
References
Barbut, S., 2002. Poultry Products Processing. An Industry Guide. CRC
Press, London.
Boerema, J.A., Penney, N., Cummings, T.L., Bell, G., 1993. Carbon
dioxide controlled atmosphere packaging of sliced ham. Int. J. Food
Sci. Tech. 28, 435–442.
Carlos, A.M.A., Harrison, M.A., 1999. Inhibition of selected microorgan-
isms in marinated chicken by pimento leaf oil and clove oleoresin. J.
Appl. Poultry Res. 8, 100–109.
Cosby, D.E., Harrison, M.A., Toledo, R.T., 1999. Vacuum or modified
atmosphere packaging and EDTA-nisin treatment to increase poultry
product shelf-life. J. Appl. Poultry Res. 8, 185–190.
de Azevedo Gomes, H., da Silva, E.D., Cardello, H.M.A.B., Cipolli,
K.M.V.A.B., 2003. Effect of gamma radiation on refrigerated
mechanically deboned chicken meat quality. Meat Sci 65, 919–926.
Dainty, R.H., 1996. Chemical/biochemical detection of spoilage. Int. J.
Food Microbiol. 33, 19–34.
 ARTICLE IN PRESS
A. Patsias et al. / Food Microbiology 23 (2006) 423–429
Dainty, R.H., Mackey, B.M., 1992. The relationship between the
phenotypic properties of bacteria from chill-stored meat and spoilage
processes. J. Appl. Bacteriol. 73, 103S–114S.
Davies, A.R., 1995. Advances in modified-atmosphere packaging. In:
Gould, G.W. (Ed.), New Methods of Food Preservation. Blackie
Academic and Professional, London, pp. 304–320.
Deak, T., Beuchat, L.R., 1996. Handbook of Food Spoilage Yeasts. CRC
Press, Boca Raton, Fl, USA.
Farber, J.M., 1991. Microbiological aspects of modified-atmosphere
packaging technology-A review. J. Food Prot. 54, 58–70.
Gardner, G.A., 1966. A selective medium for the enumeration of
Microbacterium thermosphactum in meat and meat products. J. Appl.
Bacteriol. 29, 455–460.
Geornaras, I., de Jesus, A., van Zyl, E., von Holy, A., 1998. Bacterial
populations associated with the dirty area of the South African poultry
abattoir. J. Food Prot. 61, 700–703.
Gill, C.O., 1986. The control of microbial spoilage in fresh meats. In:
Pearson, A.M., Dutson, T.R. (Eds.), Advances on Meat Research:
Meat Poultry Microbiology. Macmillan, London, pp. 49–88.
Gill, C.O., Reichel, M.P., 1989. Growth of the cold tolerant pathogens
Yeshinia enterocolitica, Aeromonas hydrophila and Listeria mono-
cytogens on high pH beef packaged under vacuum or carbon cioxide.
Food Microbiol 6, 223–230.
Hintlian, C.B., Hotchkiss, J.H., 1987. Comparative growth of spoilage
and pathogenic organisms on modified atmosphere packaged cooked
beef. J. Food Prote 50, 218–223.
Holzapfel, W.H., 1998. The Gram-positive bacteria associated with meat
and meat products. In: Davis, A., Board, R. (Eds.), Microbiology of
meat and Poultry. Blackie Academic and Professional, London,
England, pp. 35–84.
Hwang, H., Beuchat, L.R., 1995. Efficacy of selected chemicals for killing
pathogenic and spoilage microorganisms on chicken skin. J. Food
Prot. 58, 19–23.
Ingham, S.C., Escude, J.M., Cown, P., 1990. Comparative growth rates of
Listeria monocytogenes and Pseudomonas fungi on cooked chicken
loaf stored under air and two modified atmospheres. J. Food Prot 51,
177–182.
Ismail, S.A.S., Deak, T., Abd El-Rahman, H.A.M., Yassien, A.M.,
Beuchat, L.R., 2000. Presence and changes in populations of yeasts on
raw and processed poultry products stored at refrigeration tempera-
ture. Int. J. Food Microbiol. 62, 113–121.
Jimenez, S.M., Salsi, M.S., Tiburzi, M.C., Rafaghelli, R.C., Pirovani,
M.E., 1999. Combined use of acetic acid treatment and modified
atmosphere for extending the shelf-life of chilled chicken breast
portions. J. Appl. Microbiol. 87, 339–344.
International Commission on Microbiological Specifications for Foods
(ICMSF). 1986. In: Microorganisms in Foods. Sampling for Micro-
biological Analysis: Principles and Scientific Applications, Vol. 2,
second ed. University of Toronto Press, Toronto, pp. 181–196.
Kakouri, A., Nychas, G.-J.E., 1994. Storage of poultry meat under
modified atmospheres or vacuum packs: possible role of microbial
metabolites as indicator of spoilage. J. Appl. Bacteriol. 76, 163–172.
Kim, C.R., Marshall, D.L., 1999. Microbiological, colour and sensory
changes of refrigerated chicken legs treated with selected phosphates.
Food Res. Int. 32, 209–215.
Linton, M., McClements, J.M.J., Patterson, M.F., 2004. Changes in
microbiological quality of vacuum-packaged, minced chicken treated
with hydrostatic pressure. Innov. Food Sci. Emerg. Technol. 5, 151–159.
429
Mahrour, A., Caillet, S., Nketsia-Tabiri, J., Lacroix, M., 2003. The
antioxidant effect of natural substances on lipids during irradiation of
chicken legs. J. Am. Oil Chem. Soc. 80, 679–684.
Marenzi, C., 1986. Proper meat storage prevents spoilage. Poultry-Misset
6, 12–15.
Marshall, D.L., Wiese-Lehigh, P.L., Wells, J.A. and Farr, A.R.,
1991.Comparative growth of Listeria monocytogenes and Pseudominas
fluorescens on precooked chicken nuggets stored under modified
atmosphere. J. Food Prot. 54, 841-842, 851.
Marshal, D.L., Andrews, L.S., Welles, J.H., Farr, A.R., 1992. Influence of
MAP on the competitive growth of Listeria moncytogens and
Pseudomonas fluorescens on precooked chicken. Food Microbiol 9,
303–309.
Mead, G.C., Adams, B.W., 1977. A selective medium for the rapid
isolation of Pseudomonas associated with poultry meat spoilage. Brit.
Poultry Sci. 18, 661–670.
Mossel, D.A.A., Eelderink, L., Koopmans, M., Rossem, F.V., 1979.
Influence of carbon source, bile salts, and incubation temperature on
recovery of Enterobacteriaceae from foods using MacConkey-type
agars. J. Food Prot. 42, 470–475.
Nam, K.C., Ahn, D.U., 2003. Combination of aerobic and vacuum
packaging to control lipid oxidation and off-odor volatiles of
irradiated raw turkey breast. Meat Sci 63, 389–395.
Nawar, WW., 1996. Lipids. In: Fennema, O.R. (Ed.), Food Chemistry
Third ed. Marcel Dekker, New York, pp. 225–319.
Nissen, H., Sørheim, O., Dainty, R., 1996. Effects of vacuum, modified
atmospheres and storage temperature on the microbial flora of
packaged beef. Food Micro. 13 (3), 183–191.
Patsias, A., Chouliara, I., Paleologos, E.K., Savvaidis, I.N., Kontominas,
M.G., 2005. Relation of biogenic amines to microbial and sensory
changes of precooked chicken meat stored aerobically and under
modified atmosphere packaging at 41C. J. Food Protec., accepted.
Pearson, D., 1976. The Chemical Analysis of Foods. Chemical Publ. Co.,
Inc., New York, pp. 642–643.
Penney, N., Hagyard, C.J., Bell, R.G., 1993. Extension of shelf life of
chilled sliced roast beef in carbon dioxide packaging. Int. J. Sci. Tech.
28, 181–191.
Roth, L.A., Clark, D.S., 1976. Effect of Lactobacilli and carbon dioxide
on the growth of microbacterium thermosphactum on fresh beef.
Cana. J. Microbiol. 21, 629–632.
Saucier, L., Gendron, C., Gariepy, C., 2000. Shelf life of poultry meat
under modified atmosphere. Poultry Sci 79, 1851–1856.
Sawaya, W.N., Elnawawy, A.S., Abu-Ruwaida, A.S., Khalafawi, S.,
Dashti, B., 1995. Influence of modified atmosphere packaging on shelf-
life of chicken carcasses under refrigerated storage conditions. J. Food
Saf. 15, 35–51.
Steel, R.G.D., Torrie, J.H., 1980. Principles and Procedures of Statistics.
A Biometrical Approach. Mc Graw-Hill, New York.
Stiles, M.E., 1991. Scientific principles of controlled/modified atmosphere
packaging. In: Ooraikul, B., Stiles, M.E., (Eds.), Modified Atmosphere
Packaging of Food. Ellis Horwood Ltd., Chichester, England, pp.
18–25 and 118–147.
Wel, Q.K., Fang, T.J., Chen, W.C., 2001. Development and validation of
growth model for Yeshinia enterocolitica in cooked chicken meats
packages under various atmosphere packaging and stored at different
temperatures. J. Food Prot. 64 (7), 987–993.
Winger, R.J., Pope, C.G., 1976. Selection and training of panellists for
sensory evaluation of meat flavours. J. Technol. 16, 661–669.