Journal of Food Safety ISSN 1745-4565

PERSPECTIVES ON THE USE OF ESSENTIAL OILS AS

ANTIMICROBIALS AGAINST CAMPYLOBACTER JEJUNI CECT
7572 IN RETAIL CHICKEN MEATS PACKAGED IN
MICROAEROBIC ATMOSPHERE
jfs_342

37..47

DJAMEL DJENANE1, JAVIER YANGELA2, DIEGO GMEZ2 and PEDRO RONCALS2,3

1

Facult des Sciences Biologiques et des Sciences Agronomiques, Department: Biochimie et Microbiologie, Universit Mouloud Mammeri,
Tizi-Ouzou, Algeria
2
Department: Produccin Animal y Ciencia de los Alimentos, Universidad de Zaragoza. C/ Miguel Servet, 177-50013 Zaragoza, Spain

Corresponding author. TEL:

+34-976-76-15-82; FAX: +34-976-76-15-90;
EMAIL: roncales@unizar.es
Received for Publication February 23, 2011
Accepted for Publication July 28, 2011
doi:10.1111/j.1745-4565.2011.00342.x

ABSTRACT
The chemical composition of the essential oils (EOs) of Inula graveolens, Laurus
nobilis, Pistacia lentiscus and Satureja montana was analyzed using a gas
chromatography-mass spectrometry technique. The main components of EOs
obtained were, respectively, bornyl acetate, 1,8-cineole, b-myrcene and carvacrol.
EOs were screened for their ability to inhibit the growth of Campylobacter jejuni
CECT 7572 using the standard agar-disk diffusion assay. The results obtained, followed by measurements of minimal inhibitory concentrations, indicated that I. graveolens was most active (F = 53.3 mm), with the lowest MIC value against C. jejuni
(2 mL/mL). EOs were tested in chicken stored in microaerobic conditions at 3 2C,
experimentally inoculated with the pathogen at a level of 5 105 cfu/g. C. jejuni
counts in treated samples were 0.74.7 log10 cfu/g lower (P < 0.05) than the controls
throughout storage. The latter reached numbers of about 8 log10 cfu/g after 1 week.
Lipid oxidation (thiobarbituric acid reactive substances [TBARS]) and sensory
freshness odor were also determined. Samples treated with any EO had the lowest
TBARS values (P < 0.05). The presence of EOs significantly extended fresh meat
odor. The results of the bioassays, together with the chemical profile of the EOs,
support the possibility of using all EOs as potent natural preservatives to contribute
in the reduction of experimentally inoculated C. jejuni in chicken meat.

PRACTICAL APPLICATIONS
The results revealed for the first time in a chicken meat system the potential of I. graveolens, L. nobilis, P. lentiscus and S. montana EOs in inhibiting C. jejuni. This suggests the possibility that they, particularly I. graveolens, could be used as natural
preservatives in chicken meat for reducing food hazards caused by this pathogen,
which is now recognized as the leading cause of bacterial foodborne gastroenteritis.

INTRODUCTION
Campylobacter species cause campylobacteriosis in humans, a
gastrointestinal tract infection (Suzuki and Yamamoto 2009;
Wegener 2010). Campylobacter contamination of chicken
carcasses is common, and poultry meat is generally recognized to play a significant role in human Campylobacter infection (Zilbauer et al. 2008). The most important pathogenic
Journal of Food Safety 32 (2012) 3747 2011 Wiley Periodicals, Inc.

strains belong to the group of thermotolerant Campylobacter,

particularly Campylobacter jejuni (Son et al. 2007). C. jejuni
has recently overtaken Salmonella spp. as the major reported
source of foodborne bacterial diseases within the European
Union (European Food Safety Authority 2009). C. jejuni is
part of normal enteric microbiota in animals (cattle, chicken
and pigs) and can be transmitted to humans through
contaminated foods (Aslim and Yucel 2008). A positive
37

ANTIMICROBIALS FOR C. JEJUNI IN CHICKEN

correlation was observed between the number of C. jejuni

present in the caeca and the number of bacteria present on
carcasses and cut products (Reich et al. 2008). Contamination
of carcasses with C. jejuni occurs particularly during scalding,
defeathering, evisceration and chilling operations (Nauta
et al. 2009).
Several recent studies described in detail the antimicrobial
properties of some EOs against a number of relevant, foodborne, pathogenic bacteria, which may be envisaged as
natural alternatives to chemical-based antibacterial for food
safety and preservation (Bakkali et al. 2008; Solomakos et al.
2008; Djenane et al. 2011a,b,c). Despite the potential of many
common plants and EOs is considerable, knowledge of this
area and studies on their biological activities remain scarce.
Most of the data published on the antimicrobial properties of
plant EOs are fragmented and employ only basic screening
techniques. Moreover, most studies on the antimicrobial
action of plant extracts have been conducted in vitro, so that
little information exists regarding the antimicrobial activity
of EOs in food systems (Koutsoudaki et al. 2005; Oussalah
et al. 2007; Oke et al. 2009).
The main objectives of this study were (1) to determine
chemical composition of the steam-distilled EOs of Inula
graveolens, Laurus nobilis, Pistacia lentiscus and Satureja
montana by gas chromatography-mass spectrometry (GCMS); (2) to investigate the antimicrobial activity of these EOs
against the food pathogen C. jejuni by in vitro disk diffusion
methods, as well as to determine their minimum inhibitory
concentration (MIC); (3) to study their antimicrobial effect
in a chicken meat system; and (4) to investigate the effect of
EO addition on the sensory properties of meat throughout
storage.

MATERIALS AND METHODS

Plant Material and Essential Oils Extraction
The aerial parts of I. graveolens and L. nobilis were collected at
Tizi-Ouzou province (Algeria), from March to July 2009 and
authenticated by the Department of Biology, University
Mouloud Mammeri of Tizi-Ouzou (Algeria). The whole
fresh plants were then extensively washed with distilled
water (20C) to remove epiphytic hosts normally found on the
surface and were dried in the darkness at 25C. Only the leaves
were recuperated for subsequent extraction to obtain their
EOs. EOs from P. lentiscus and S. montana were purchased
from Florame Aromathrapie (St Rmy de Provence, France).
Both EOs were certified by Ecocert SAS F32600 (France) and
were considered 100% pure and natural, obtained from
Mediterranean biological culture.
The I. graveolens and L. nobilis EOs were obtained from
dried leaves and plant parts by steam hydrodistillation in a
Clevenger-type apparatus for 3 h (Groupe Pharmaceutique
38

D. DJENANE ET AL.

SAIDAL, Filiale Biotic, Algiers, Algeria). The EOs obtained

were separated from water and dried over anhydrous sodium
sulphate (Na2SO4). The isolated EOs were preserved in darkness in a sealed vial at 1C until its analysis or its use in bioassays.

Analysis of Essential Oils

GC Analysis. GC analyses of EOs obtained from dried
material were performed using a Hewlett Packard 6890
gas chromatograph (Hewlett-Packard Company, Palo Alto,
CA) equipped with a flame ionization detector (FID) and
a Stabilwax (polyethylene glycol) column (30 m 0.32 mm
internal diameter, 1-mm film thickness; Centre de Recherche
en Analyses Physico-Chimiques [CRAPC]; USTHB, Algiers,
Algeria).
The operating conditions were as follows: injector and
detector temperatures, 250 and 280C, respectively; carrier
gas, N2 at a flow rate of 1 mL/min; oven temperature
program, 3 min isothermal at 50C, raised at 2C/min to 220C,
and finally held isothermal for 15 min. The identities of the
separated components on the polar column were determined
by comparing their retention indices relative to aliphatic
hydrocarbons injected under the above temperature program
with literature values measured on columns with identical
polarities.
GC-MS Analysis. The Gas chromatography-mass spectrometry (GC-MS) analysis was performed using a HewlettPackard 6890 series GC systems coupled to a quadrupole
mass spectrometer (model HP 5973) equipped with an
HP5 MS capillary column (5% phenyl methylsiloxane,
30 m 0.25 mm, 0.25-mm film thickness; CRAPC, USTHB).
For GC-MS detection, an electron ionization system with
ionization energy of 70 eV was used over a scan range of
30550 atomic mass units. Helium was the carrier gas, at a
flow rate of 0.5 mL/min. Injector and detector MS transfer
line temperatures were set at 250 and 280C, respectively; the
temperature of the ion source was 230C. Column temperature was initially kept at 60C for 8 min, then gradually
increased to 280C at 2C/min, and finally held isothermal for
30 min. The volume of injections was 0.2 mL of a hexaneoil
solution, injected in the splitless mode. The identity of the
components was assigned by matching their spectral data
with those detailed in the Wiley 7N, National Institute of
Standards and Technology (NIST) 02 and NIST 98 libraries.
The results were also confirmed by the comparison of their
retention indices, relative to C7-C29 n-alkanes assayed under
GC-MS in the same conditions as the oils. Some structures
were further confirmed by available authentic standards
analyzed under the same conditions described earlier. The
percentage composition of the oils was computed by the
normalization method from the GC peak areas, calculated
as the mean value of two injections from each EO.
Journal of Food Safety 32 (2012) 3747 2011 Wiley Periodicals, Inc.

D. DJENANE ET AL.

Antibacterial Activity Assays

Bacterial Strain and Culture Conditions. The bacterial
strain of gram-negative C. jejuni studied was provided by the
Spanish Type Culture Collection. Strain used was C. jejuni
CECT 7572. Bacterial strain was grown in Bolton broth
(CM0983) supplemented with 5% laked horse blood
(SR0048; BioMrieux, Marcy lEtoile, France) and selective
supplement (SR0183) at 42C in microaerobic conditions (5%
O2, 10% CO2 and 85% N2). One milliliter of stock culture was
standardized through two successive 24-h growth cycles at
42C in 9 mL of brainheart infusion broth (BHIB; Oxoid,
Basingstoke, U.K.). After 48 h, 100 mL of the suspension was
then inoculated in fresh BHIB and incubated at 42C for 12 h
to obtain a working fresh culture containing about
5 105 cfu/mL, determined by measuring transmittance at
600 nm (Spectrophotometer: Spectronic 20 Bausch & Lomb
U.K. Ltd., Kingston-Upon-Thames, Surrey, U.K.). The strain
was maintained frozen (-80C) in cryovials (Cryobanks, Mast,
Merseyside-Liverpool, U.K.) containing an antifreezing agent
to preserve the viability of the cells during storage and were
subcultured every antibacterial test.

Screening of EO. Screening of EOs for antibacterial activity

was done by the disk diffusion method. Petri dishes were prepared by pouring 20 mL of Mueller Hinton (MH) agar
medium and allowed to solidify.Plates were dried for 30 min in
a biological safety cabinet with vertical laminar flow and
0.1 mL of standardized inoculums suspension was poured and
uniformly extended. The inoculums were allowed to dry for
5 min. To prepare the stock solution of the samples, the pure
EOs were dissolved in 5% (v/v) dimethyl sulfoxide (DMSO;
Sigma-Aldrich Qumica S.A., Madrid, Spain). Then sterile
filter paper disk (6-mm diameter, Filter LAB ANOIA, testing
paper, Barcelona, Spain) was impregnated with 05 mL EO,
using a capillary micropipette (Finnpipette, Thermo Fischer
Scientific Inc., Helsinki, Finland). The dishes were left for
15 min at room temperature to allow the diffusion of the EO,
and then they were incubated at 42C for 2448 h in microaerobic conditions. At the end of the period, the diameter of the
clear zone around the disk was measured with a caliper (Wiha
dialMax, Schonach, Germany, ESD-Uhrmessschieber, CH)
and expressed in millimeters (disk diameter included) as its
antimicrobial activity. The sensitivity to the different EOs
was classified by the diameter of the inhibition halos as follows:
not sensitive (-) for diameter less than 8 mm; sensitive (+)
for diameter 914 mm; very sensitive (++) for diameter
1519 mm and extremely sensitive (+++) for diameter larger
than 20 mm (Ponce et al. 2003). Negative controls were prepared using the same solvent employed to dissolve the samples.
Standard reference antibiotic, gentamicin (10 mg/disk; Sigma
Aldrich Qumica S.A.), was used as positive control to
Journal of Food Safety 32 (2012) 3747 2011 Wiley Periodicals, Inc.

ANTIMICROBIALS FOR C. JEJUNI IN CHICKEN

determine the sensitivity of the tested microorganism. Each

assay in this experiment was replicated three times.
Microdilution Assays. The MIC values were also studied
for the target bacterium, which was determined as sensitive to
the EOs in disk diffusion assay, as described in the earlier
section. The inoculum of C. jejuni was prepared from 12-h
broth cultures and suspension was adjusted to 0.5 McFarland
standard turbidity to give a final density of 3 105 cfu/mL.
I. graveolens, L. nobilis, P. lentiscus and S. montana EOs dissolved in 0.5% DMSO were first diluted to the highest concentration (32 mL/mL) to be tested, and then serial twofold
dilutions were made in a concentration range from 32 to
0.3125 mL/mL in 10-mL sterile test tubes containing MH
broth. MIC values of all EOs against C. jejuni were determined based on a microwell dilution method. The 96-well
plates (Iwaki brand, Asahi Techno Glass, Funabashi, CHB,
Japan) were prepared by dispensing into each well 95 mL of
MH broth and 5 mL of the inoculum. A 100-mL aliquot from
all EOs extracts initially prepared at the concentration of
32 mL/mL was added into the first wells. Then, 100 mL from
their serial dilutions was transferred into consecutive wells.
The last well containing 195 mL of nutrient broth without
compound and 5 mL of the inoculums on each strip was used
as negative control. The final volume in each well was 200 mL.
After incubation at 42C for 1824 h under microaerobic conditions (5% O2, 10% CO2 and 85% N2; Genbox microaer,
BioMrieux) in 2.5-L anaerobic jars (Oxoid, Basingstoke,
U.K.) with agitation; the wells were then examined for evidence of growth and MICs (mL/mL) values were determined
as the lowest EO concentration that inhibited visible growth
of the tested microorganism, which was indicated by absence
of turbidity. The negative control was set up with DMSO in
amount corresponding to the highest quantity present in the
test solution (0.5%). The tests were performed in duplicate
and repeated twice.

Inhibitory Effect of the Essential Oils against

C. jejuni Inoculated in Chicken Meat
Preparation of Chicken Meat. Chicken muscles were
aseptically cut by means of a sterile meat knife from chicken
carcasses at about 6 h postslaughter (local supermarket,
Alcampo, Zaragoza, Spain) and transported to the laboratory
under refrigerated conditions within 30 min. After the aseptic
removal of the outer surface, meat was aseptically prepared by
means of a sterile steel meat preparation.
Treatment of Chicken Meat. Prior to meat inoculation
with C. jejuni and the addition of EOs, chicken muscles were
also examined for any contamination by bacteria or the tested
pathogen (results not shown). To evaluate the antimicrobial
activity of all EOs in a chicken meat system, a sufficient
39

ANTIMICROBIALS FOR C. JEJUNI IN CHICKEN

amount of fresh chicken meat was prepared following good

practices and was tested using the twofold MICs value found
for all EOs. After the aseptic removal of the outer surface. A
total of 80 meat samples were obtained. Two individual duplicates of each sample were performed in all cases. For microbiological study, meat samples (40 100 2 g) were placed
individually in stomacher bags and inoculated with strain of
C. jejuni at a level of 3 105 cfu/g. The inoculated samples
were homogenized to ensure proper distribution of the
pathogen. Following homogenization, the individual EO was
added to the inoculated samples. Addition of EO was done
at twofold MIC values (0.20.6%, respectively). To attain
uniform distribution of the added compounds, treated meat
samples were further homogenized, as previously described.
All stomacher bags with samples from all treatments were
wrapped in a pouch made of a polyethylene and polyamide
laminate (Sidlaw Packaging-Soplaril, Barcelona, Spain) of
water vapor permeability 57 g/m2/24 h at 23C and oxygen
permeability 4050 mL/m2/24 h atm at 23C. The pouch was
filled with 1.5 L of a gas mixture of 5% O2 + 10% CO2 + 85%
N2, supplied by Abell-Linde S.A. (Barcelona, Spain), thermosealed and stored in the dark at 3 2C for 8 days.
Microbial analyses of samples for populations of C. jejuni
were carried out at 2-day intervals up to the eighth day of
refrigerated storage. However, for the oxidation study, the
remainder (40 100 2 g) meat samples were placed into
polystyrene trays. The 40 portions were divided into five
groups of eight. The first group (control) was sprayed with
sterile distilled water, using 1.5 mL of solution to 100 g of
meat. The final four groups were similarly sprayed respectively with each EO solution, respectively. Samples from all
treatments were wrapped and stored under aerobic conditions at 3 2C for 8 days.
On days 2, 4, 6 and 8 of storage, two packs containing each
sample from each group were opened. One pouch from each
of the set was used for microbiological sampling, while the
other two were used for sensory analysis and for chemical
analyses.
Bacterial Enumeration. Microbiological analyses of
samples for populations of C. jejuni were carried out at
2-day intervals up to the eighth day of refrigerated storage
(3 2C). At each sampling time, samples (25 g) of chicken
muscle in the stomacher bags were aseptically added with
225 mL of sterile peptone water. The contents were macerated in the stomacher (Stomacher 400-Circulator. Seward.
Worthing, U.K.) for 1 min at room temperature. Resulting
slurries were serially diluted (1:10) in sterile peptone water.
Sample dilutions (0.1 mL) were spread plated on appropriate media in duplicate. The selective media used for
numeration of C. jejuni was modified charcoal cefoperazone
deoxycholate agar (mCCDA, Oxoid; CM0739 + SR0155).
The mCCDA plates were incubated for 48 h at 42C in 2.5-L
40

D. DJENANE ET AL.

gas jars with CampyGen microaerophilic generating gas

packs (BioMrieux). Triplicate sets of plates were prepared
on each occasion and each experiment repeated three times.
Counts were expressed as the log10 of colony forming units
(cfu) per gram.
Thiobarbituric Acid Reactive Substances. Lipid oxidation was measured in triplicate by the 2-thiobarbituric acid
(TBA) method of Pfalzgraf et al. (1995). Meat samples of 10 g
were taken and mixed with 20 mL trichloroacetic acid (10%),
using an Ultra-Turrax T25 macevator (Janke & Kunkel,
Staufen, Germany). Samples were centrifuged at 2,300 g for
30 min at 5C; supernatants were filtered through quantitative
paper (MN 640 W, Machinery-Nagel GmbH & Co. KG,
Dren, Germany). 2 mL of the filtrate was taken and mixed
with 2 mL of thiobarbituric acid (20 mM); tube contents
were homogenised and incubated at 97C for 20 min in
boiling water. Absorbance was measured at 532 nm. The concentration of the samples was calculated using a calibration
curve. TBARS values were expressed as milligrams of malonaldehyde per kilogram of sample.
Sensory Analysis. Samples of chicken meat were evaluated
for freshness odor by a sixth-member trained panel.
Panelists were selected among students and staff of the department and trained according to the American Meat Science
Association guidelines (AMSA 1995). The attribute freshness
odor was evaluated using a 5-point scale. Odor scores referred
to the intensity of freshness odor, inversely associated to meat
spoilage: 5 = very desirable odor, 4 = desirable odor,
3 = slightly desirable odor, 2 = moderately undesirable odor
and 1 = very undesirable odor (Djenane et al. 2001).
Statistical Analysis. Variance analyses were used to test
the significant difference among the results from the antibacterial assays, sensory and chemical analysis (SPSS 10.0 software package, SPSS Inc., Chicago, IL, USA; SPSS 1995).
Means and standard errors (SE) of the samples were calculated. Three replicates were performed for each treatment.
Differences between means were tested through least square
difference and values of P < 0.05 were considered significantly different.

RESULTS AND DISCUSSION

Chemical Composition of the EOs
Steam distillation is the most commonly used method for
producing EOs on a commercial basis. The average values of
hydrodistillation extraction yields of plant EOs from I. graveolens and L. nobilis were found to be 0.15 and 0.084% (v/w),
respectively. The results obtained in the qualitative and quanJournal of Food Safety 32 (2012) 3747 2011 Wiley Periodicals, Inc.

D. DJENANE ET AL.

ANTIMICROBIALS FOR C. JEJUNI IN CHICKEN

TABLE 1. MAIN CONSTITUENTS (%) OF THE ESSENTIAL OILS OF INULA, LAUREL, PISTACIA AND SATUREJA SPECIES, AS IDENTIFIED BY GAS
CHROMATOGRAPHY-MASS SPECTOMETRY ANALYSIS
Retention time (min)
1
5.177
2
5.320
3
5.527
4
5.932
5
6.640
6
6.844
7
7.287
8
7.335
9
7.763
10
8.232
11
8.577
12
8.742
13
8.773
14
9.130
15
9.544
16
9.887
17
10.034
18
10.392
19
11.349
20
12.020
21
14.725
22
15.377
23
16.063
24
16.854
25
19.982
26
20.877
27
22.330
28
23.330
29
23.854
30
24.740
31
26.310
32
26.944
33
28.563
34
30.215
35
32.135
36
33.802
37
34.759
38
35.040
39
35.716
40
36.428
41
39.611
Monoterpenes hydrocarbons
Oxygenated monoterpenes
Sesquiterpene hydrocarbons
Oxygenated sesquiterpenes
Total identified

Compound

Inula graveolens

Laurus nobilis

Tricyclene
a-Thujene
a-Pinene
Camphene
Sabinene
b-Pinene
Myrcene
b-Myrcene
a-Phellandrene
a-Terpinene
p-Cymene
Limonene
1,8-cineole
trans-b-ocimene
cis-b-ocimene
Isoamyle butyrate
g-terpinene
Trans-4-thujanol
Terpinolene
Linalool
Menthone
Borneol
Terpinen-4-ol
a-Terpineol
Thymol methyl ether
Farnesol
Bornyl acetate
Carvacrol
Thymol
Myrtenyl acetate
Terpenyl acetate
Eugenol
Geranyl acetate
Caryophyllene
a-caryophyllene
Germacrene-D
g-elemene
a-cadinene
b-bisabolene
d-cadinene
Caryophyllene oxide

8.17
1.05

0.64
0.70
7.15
0.81
8.12
3.17
0.65
0.80

0.58
1.03
2.41

21.04

1.52

40.85

1.27

2.05
13.22%
65.82%
3.27%
5.05%
84.62%

Pistacia lentiscus

40.25

0.85

4.95

1.87
2.16

10.15
2.10

0.71

25.86%
49.23%
11.98%

87.08%

Satureja montana

0.73
0.79
0.51

5.54
3.15

5.10

15.18
3.83
2.78
1.64

15.02

1.68
0.51
4.10

2.21

6.41
2.97

1.88

0.73

4.03
0.84
0.87

0.56

1.80

53.74%
31.34%
13.62%

98.7%

0.96
1.04

1.33
11.77
0.64

0.92

6.72
1.05

1.97
0.59
1.75
1.04

0.95
4.10

29.19
15.41

0.53
5.38

0.75

0.87
0.23
0.98
43.76%
12.54%
31.63%
6.87%
94.8%

Only components percentage at >0.5 were represented.

titative analyses according to their elution order on a HP5 MS

capillary column are shown in Table 1.
From the oil of I. graveolens, 30 constituents were identified, representing 87.36% (area percent) of the total oil,
among which bornyl acetate (40.85%), borneol (21.04%),
camphene (8.17%) and 1,8-cineole (2.41%) were the major
Journal of Food Safety 32 (2012) 3747 2011 Wiley Periodicals, Inc.

compounds. The most abundant chemical category were the

oxygenated monoterpenes (65.82%), followed by monoterpene hydrocarbons (13.22%), oxygenated sesquiterpenes
(5.05%) and sesquiterpene hydrocarbons (3.27%).
Forty-seven constituents have been identified from the oil
of L. nobilis, representing 87.08% of the total oil, among
41

ANTIMICROBIALS FOR C. JEJUNI IN CHICKEN

D. DJENANE ET AL.

f (mm)a*

Campylobacter
jejuni CECT 7572

I. graveolens

L. nobilis

P. lentiscus

S. montana

Gentamicine

53.3 9.0a*

37.3 5.5b

25.3 1.52c

25.8 0.2c

21.3 2.6c

All tests were performed in triplicate.

Values followed by the same letter, are not significantly different (P > 0.05).
* af: Inhibition zone in diameter around the disks impregnated with essential oils. The diameter
(6 mm) of the disk is included.

which 1,8-cineole (40.25%), terpenyl acetate (10.15%), sabinene (8.12%), a-pinene (7.15%) and linalool (5%) were
found as the most abundant components. Oxygenated
monoterpenes (49.23%) were the most abundant chemical
category, followed by monoterpene hydrocarbons (25.86%)
and sesquiterpene hydrocarbons (11.98%).
Fifty-seven constituents were detected from the oil of
P. lentiscus, representing 98.7% of the total oil, among which
b-myrcene (15.18%), 1,8-cineole (15.02%), terpinen-4-ol
(6.41%) and a-b-pinene (5.54% and 5.10% respectively)
were the major ones. The most abundant chemical category
were monoterpene hydrocarbons (53.74%), followed by oxygenated monoterpenes (31.34%) and sesquiterpene hydrocarbons (13.62%).
From the oil of S. montana, 44 compounds were identified,
representing 94.8%, among which carvacrol (29.19%),
thymol (15.41%), p-cymene (11.77%) and g-terpinene
(6.72%) were the main constituents. The oil was rich in
monoterpene hydrocarbons (43.76%), followed by the sesquiterpene hydrocarbons (31.63%), the oxygenated monoterpenes (12.54%) and the oxygenated sesquiterpenes
(6.87%). Our study supports the view that bornyl acetate and
1,8-cineole are major components of the EOs of I. graveolens
and L. nobilis, respectively, of Algerian origin.
In agreement with our results, Bokadia et al. (1986) and
Barla et al. (2007) found that the characteristic compounds of
the genus Inula and Laurus were monoterpenes and sesquiterpenes. The profile of the volatile oils obtained was also in
agreement with values reported from other countries. L. nobilis oil from Buenos Aires (Argentina) contains sabinene
(8%), 1,8-cineole (42.75%), linalool (13.23%) and a-terpinyl
(7.6%) (Lira et al. 2009). However, it has been found that
P. lentiscus EO from Greece was characterized by a high
monoterpene hydrocarbon fraction (4568.3%), followed by

oxygenated monoterpenes (13.323.1%) and sesquiterpene

hydrocarbons (9.228.1%) (Chryssavgi et al. 2008). Similar
findings have been reported by other authors (Zrira et al.
2003). In agreement with our results, previous investigations
on the chemical composition of the EO of Satureja species
indicated that it contains carvacrol and thymol as major components (Baser 2002).
The different qualitative and quantitative chemical compositions of these EOs with respect to previous investigations
could be related first and foremost to the different environmental conditions, genetics (degree of hybridization), geographical origin and harvest period (Slavkovska et al. 2001;
Mastelic and Jerkovic 2003).

Antimicrobial Activity (Disk Assay)

According to the results given in Table 2, the oil of I. graveolens showed the largest inhibitory effects against target bacteria (F = 53 mm), which was followed by that of L. nobilis
(37.3 5.5 mm; P < 0.05), while the lowest were those of
P. lentiscus and S. montana (25.3 1.52 and 25.8 0.2 mm,
respectively; P < 0.05). The average zone of inhibition of the
antibiotic gentamicin used as a positive control against the
same target bacteria was of only 21 2.6 mm. Those results
were consistent with those of the microdilution broth assay
(Table 3), since the EO of I. graveolens exhibited a MIC value
of 0.2% and all other three EOs reached only a MIC of 0.6%.
Smith-Palmer et al. (2001) reported similar results regarding the effect of oils of bay and thyme on C. jejuni. The same
authors recorded MICs close to this value when they tested the
antimicrobial properties of the EOs of 21 plants and two
essences against C. jejuni. Recently, Nannapaneni et al. (2009)
found that seven orange oil fractions showed a large inhibitory
effect on both C. jejuni and C. coli, exhibiting zones of

Minimal inhibitory concentration (%)*

Campylobacter
jejuni CECT 7572

TABLE 2. ANTIBACTERIAL ACTIVITY OF THE

EOs FROM INULA GRAVEOLENS, LAURUS
NOBILIS, PISTACIA LENTISCUS AND SATUREJA
MONTANA, USING PAPER DISK DIFFUSION
METHOD, EXPRESSED BY DIAMETER (mm) OF
INHIBITION ZONE (INCLUDING THE DISK
DIAMETER, 6 mm)

I. graveolens

L. nobilis

P. lentiscus

S. montana

0.2 0.02

0.6 0.05

0.6 0.02

0.6 0.04

TABLE 3. MINIMAL INHIBITORY

CONCENTRATIONS VALUES FROM INULA
GRAVEOLENS, LAURUS NOBILIS, PISTACIA
LENTISCUS AND SATUREJA MONTANA
ESSENTIAL OILSS USING BROTH
MICRODILUTION METHOD

All tests were performed in triplicate.

* Minimal inhibitory concentrations values expressed by % (v/v).

42

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D. DJENANE ET AL.

inhibition of up to 80 mm. On the other hand, C. jejuni

was inhibited by bergamot (F = 23 mm, MIC > 4%),
lemon (F = 18 mm, MIC > 4%), linalool (F > 90 mm,
MIC = 0.06%) and linalool vapor (Fisher and Phillips 2006).
Many reports confirm that the EOs of plant species studied in
our work are among the most potent antimicrobial agents (De
Corato et al. 2010; Zhao et al. 2010; Djenane et al. 2011b,c).
The antibacterial effect of all four EOs may be attributed to
their high content of compounds with known antimicrobial
activity. The phenolic components are chiefly responsible for
the antibacterial properties of EOs, but, aldehydes and alcohols also exhibit antimicrobial effects (Fitzgerald et al. 2003).
These components are able to disintegrate the outer membrane of gram-negative bacteria, releasing lipopolysaccharides and increasing the permeability of the cytoplasmic
membrane to adenosine triphosphate (Marino et al. 2001).
The strength and spectrum of activity is known to vary
according to the Gram type of the target bacteria; grampositive bacteria were generally more sensitive to the effects of
the EOs than gram-negative bacteria (Delaquis et al. 2002).
The antimicrobial activities of the EOs are difficult to correlate to a specific compound due to their complexity and
variability. Synergism has been observed between various
components of the same EO (Burt 2004). Some studies have
concluded that whole EOs have a greater antibacterial activity
than the major components individually (Gill et al. 2002;
Mourey and Canillac 2002). Nevertheless, some researchers
reported that there is a relationship between the chemical
composition of the most abundant components in the EO
and the antimicrobial activity. For example, 1,8-cineole
(abundant in Algerian L. nobilis EO tested in this study) is
well known for its antimicrobial potential (Pattnaik et al.
1997). Lis-Balchin and Deans (1997) showed that EOs containing large amounts of 1,8-cineole are better antilisterial
agents than EOs that do not contain 1,8-cineole. The antimicrobial effects of borneol (abundant in tested Algerian I. graveolens EO) were also reported by Dorman and Deans (2000).
As a result of these findings, the higher antimicrobial activity of I. graveolens EO could be attributed to this particular
chemotype, characterized by its complexity, with bornyl
acetate, borneol and camphene being the most abundant,
which have a well-documented antibacterial and antifungal
potential (Marino et al. 2001; Holley and Patel 2005).
It has been indicated that the results of the disk diffusion
method show a highly significant correlation with MIC
values (Oke et al. 2009). However, another report found that
the MIC values of EOs on foodborne pathogens were lowly
correlated with disk diffusion results (Djenane et al. 2011a).
This can be explained by the fact that the sensitivity depends
on the type of bacteria, the type, composition and concentration of the EO, the solubility in media, and seasonal and
intraspecific compositional variations. Generally, EOs produced from herbs harvested during or immediately after
Journal of Food Safety 32 (2012) 3747 2011 Wiley Periodicals, Inc.

ANTIMICROBIALS FOR C. JEJUNI IN CHICKEN

flowering possess the strongest antimicrobial activity

(Marino et al. 1999).

Antimicrobial Activity in Chicken Meat

Figure 1 depicts C. jejuni growth inhibition in chicken meat
by the four tested EOs, applied at twofold MIC values, compared with the control. It appeared to be evident that, despite
some significant differences among them in a first phase of
bacterial inactivation, all EOs reached similar inhibition
activities after 4 days of storage. The initial population of 5.6
log10 cfu/g of C. jejuni increased to 8.14 log10 cfu/g by the end
of storage in untreated samples. A reduction of 3.08, 3.50,
3.08 and 2.08 log10 cfu/g was recorded in 4 days of storage,
respectively by I. graveolens, L. nobilis, P. lentiscus and S. montana. Four days later (at day 8), reductions of 6.94, 6.14, 5.94
and 5.94 log10 cfu/g, were reached respectively by I. graveolens, L. nobilis, P. lentiscus and S. montana. As far as we are
aware, the antibacterial effect of these EOs against C. jejuni in
chicken meat had not yet been reported.
These results for the inhibition of C. jejuni in a meat
system were in good agreement with prior in vitro results,
as well as with the MIC calculated values. However, the
expected higher effect of I. graveolens was not confirmed.
Several studies have reported the effect of a food matrix on
microbial resistance to EOs, but no one of them quantified
it nor explained the mechanism, although some suggestions
have been made.
Little research has been carried out on the effect of Inula,
Laurus, Pistacia and Satureja EOs against C. jejuni. Further

FIG. 1. SURVIVAL CURVES OF CAMPYLOBACTER JEJUNI BY INULA

GRAVEOLENS, LAURUS NOBILIS, PISTACIA LENTISCUS AND SATUREJA
MONTANA ESSENTIAL OILS AT TWOFOLD MINIMUM INHIBITORY
CONCENTRATION VALUES IN CHICKEN MEAT STORED AT 3 2C
UNDER MICROAEROBIC CONDITIONS
() Control; () S. montana; () P. lentiscus; ( ) I. graveolens; ()
L. nobilis. The error bars represent standard deviation.

43

ANTIMICROBIALS FOR C. JEJUNI IN CHICKEN

D. DJENANE ET AL.

TBA Reactive Substances and

Sensory Analysis

FIG. 2. TBARS (MILLIGRAMS OF MALONALDEHYDE PER KILOGRAM OF

MEAT) IN CHICKEN TREATED WITH ESSENTIAL OILS AND STORED AT
3 2C UNDER AEROBIC CONDITIONS
() Control; () S. montana; () P. lentiscus; ( ) I. graveolens; ()
L. nobilis
The error bars represent standard deviation.

reviews of existing literature revealed that other herbs were

also effective in inhibiting C. jejuni. Lee et al. (2004) found
that an aqueous leek extract was very effective against Campylobacter species. Previous studies using other EOs have shown
antibacterial activity (Lis-Balchin and Deans 1997; Zhao et al.
2010).
The effect of combining these EOs remains to be elucidated. Nevertheless, Djenane et al. (2011c) reported that the
combination of P. lentiscus and S. montana at low concentrations (0.03%) exhibited a higher activity against Listeria
monocytogenes than individual EOs applied at higher concentrations (0.06%). Synergism between EOs and other parameters in antimicrobial action must be therefore considered.
Thus, further research is needed to evaluate the effectiveness
of combined EOs in this and other food systems, as well as by
using active packaging (Camo et al. 2008), in order to assess
their performance as natural antimicrobial agents in food
preservation and safety.

Figure 2 shows the results for TBARS indices during storage

of chicken meat under microaerobic conditions. Lipid oxidation increased slightly with increasing time in all samples
until the fourth day of storage. Significant differences
(P < 0.05) were evident among samples thereafter, depending
on the presence or not of the EOs. Untreated samples of
chicken meat showed the highest values of TBARS (P < 0.05);
they were well above 4 at day 8 of storage. On the other hand,
samples treated with any of the EOs had significantly lower
values (P < 0.05); they did not reach 1 mg malonaldehyde/kg,
even after 8 days of storage. Martnez et al. (2006) found that a
TBARS index of 1.5 mg/kg is closely related to perceptible and
unacceptable off-odor of meat by trained panel. SnchezEscalante et al. (2003) and Djenane et al. (2002) reported that
beef patties and beef steaks, respectively, treated with rosemary exhibited lower TBARS values and metmyoglobin
percentage than untreated samples during storage.
The protective effect of meat against lipid oxidation by
antioxidant treatment had been already reported by Djenane
et al. (2003) and Camo et al. (2011). The antioxidant activity
of EOs may be ascribed to the same chemical components
than those found in antioxidants. The polyphenols found in
the EOs of I. graveolens, L. nobilis, P. lentiscus and S. montana
may act as radical scavenging agents (Sharififar et al. 2007;
Tepe et al. 2007).
With a view to assess the possible effect of added EOs on
the sensory properties of meat, a sensory evaluation of meat
odor was carried out throughout a storage period of 8 days.
Sensory scores for freshness odor are summarized in Table 4.
Results showed that fresh meat odor intensity decreased
throughout storage in all samples, though not at the same
rate. Untreated samples were given scores below 3 by the panelists after 6 days of storage, whereas samples treated with any
of the EOs were given scores above 3, even at the end of the
storage period. Thus, any of the EOs significantly (P < 0.05)
extended fresh chicken odor, reaching a maximum of 3.5,
which may be considered as acceptable, after 8 days of storage.

TABLE 4. EFFECT OF EOs ON FRESHNESS ODOR SENSORY SCORES (MEAN STANDARD DEVIATION) OF CHICKEN MEAT STORED AT 3 2C
Days of storage
Treatment

Freshness odors*

5.00 00
5.00 00a
5.00 00a
5.00 00a
5.00 00a

Control
Inula graveolens (0.40%)
Laurus nobilis (1.2%)
Pistacia lentiscus (1.2%)
Satureja montana (1.2%)

2
a

4.83 0.4
5.00 00a
5.00 00a
4.83 0.4a
5.00 0.3a
a

3.66 0.5
4.66 0.5b
4.50 0.5b
4.83 0.4b
4.66 0.5b
a

2.83 0.4
4.00 00b
3.66 0.8bc
4.00 0.5b
3.33 0.5bc
a

1.50 0.5a
3.00 00b
3.00 0.6b
3.50 0.5b
3.50 0.5b

Mean values in the same column and relating to freshness odor are significantly different when accompanied by different letters (P < 0.05).
* 5 = Very desirable odor; 4 = Desirable odor; 3 = Slightly desirable odor; 2 = Moderately undesirable odor; and 1 = Very undesirable odor.

44

Journal of Food Safety 32 (2012) 3747 2011 Wiley Periodicals, Inc.

D. DJENANE ET AL.

It may, therefore, be concluded that from a practical point of

view, the use of EOs for chicken preservation is clearly advantageous since besides contributing to shelf-life extension; it
contributes to maintaining a pleasant fresh meat odor.

CONCLUSION
Our data support the possible use of I. graveolens, L. nobilis,
P. lentiscus and S. montana EOs, particularly that from I. graveolens, for the preservation of chicken meat. By using this
method, chicken meat can be stored in a modified atmosphere assuring a low risk associated to Campylobacter, at the
same time that lipid oxidation is inhibited, giving rise to a
higher sensory quality.

ACKNOWLEDGMENTS
The authors are grateful to Ministerio de Asuntos Exteriores
y Cooperacin of Spain (AECID) and Ministre de
lEnseignement Suprieur et de la Recherche Scientifique of
Algeria for financial assistances to this work within the Programa de Cooperacin Interuniversitaria e Investigacin
Cientfica PCI/MED Algeria-Spain (grant ALI A/011170/07;
A/019342/08; A/023365/09; A/033506/10) and CNEPRU
(F00520090025), respectively.
REFERENCES
AMSA 1995. Research Guidelines for Cookery, Sensory Evaluation,
and Instrumental Tenderness Measurements of Fresh Meat,
American Meat Science Association and National Live Stock
and Meat Board, Chicago, IL.
ASLIM, B. and YUCEL, N. 2008. In vitro antimicrobial activity of
essential oil from endemic Origanum minutiflorum on
ciprofloxacin-resistant Campylobacter spp. Food Chem. 107,
602606.
BAKKALI, F., AVERBECK, S., AVERBECK, D. and IDAOMAR, M.
2008. Biological effects of essential oils a review. Food Chem.
Toxicol. 46, 446475.
BARLA, A., TOPU, G., KSZ, S., TMEN, G. and
KINGSTON, D.G.I. 2007. Identification of cytotoxic
sesquiterpenes from Laurus nobilis L. Food Chem. 104,
14781484.
BASER, K.H.C. 2002. Aromatic biodiversity among the flowering
plant taxa of Turkey. Pure Appl. Chem. 74, 527525.
BOKADIA, M.M., MACLEOD, A.J., MEHTA, S.C., MEHTA, B.K.
and PATEL, H. 1986. The essential oil of Inula racemosa.
Phytochemistry 25, 28872888.
BURT, S. 2004. Essential oils: Their antibacterial properties and
potential applications in foods a review. Int. J. Food
Microbiol. 94, 223253.
CAMO, J., BELTRN, J.A. and RONCALS, P. 2008. Extension of
the display life of lamb with an antioxidant active packaging.
Meat Sci. 80, 10861091.

Journal of Food Safety 32 (2012) 3747 2011 Wiley Periodicals, Inc.

ANTIMICROBIALS FOR C. JEJUNI IN CHICKEN

CAMO, J., LORS, A., DJENANE, D., BELTRN, J.A. and

RONCALS, P. 2011. Display life of beef packaged with an
antioxidant active film as a function of the concentration of
oregano extract. Meat Sci. 88, 174178.
CHRYSSAVGI, G., VASSILIKI, P., ATHANASIOS, M., KIBOURIS,
T. and MICHAEL, K. 2008. Essential oil composition of
Pistacia lentiscus L. and Myrtus communis L.: Evaluation of
antioxidant capacity of methanolic extracts. Food Chem. 107,
11201130.
DE CORATO, U., MACCIONI, O., TRUPO, M. and DI SANZO,
G. 2010. Use of essential oil of Laurus nobilis obtained by means
of a supercritical carbon dioxide technique against post harvest
spoilage fungi. Crop Prot. 29, 142147.
DELAQUIS, P.J., STANICH, K., GIRARD, B. and MAZZA, G.
2002. Antimicrobial activity of individual and mixed fractions
of dill, cilantro, coriander and eucalyptus essential oils. Int. J.
Food Microbiol. 74, 101109.
DJENANE, D., SNCHEZ-ESCALANTE, A., BELTRN, J.A. and
RONCALS, P. 2001. Extension of the retail display life of fresh
beef packaged in modified atmosphere by varying lighting
conditions. J. Food Sci. 66, 181186.
DJENANE, D., SNCHEZ-ESCALANTE, A., BELTRN, J.A. and
RONCALS, P. 2002. Ability of a-tocopherol, taurine and
rosemary, in combination with vitamin C, to increase the
oxidative stability of beef steaks packaged in modified
atmosphere. Food Chem. 76, 407415.
DJENANE, D., SNCHEZ-ESCALANTE, A., BELTRN, J.A. and
RONCALS, P. 2003. The shelf-life of beef steaks treated with
dL-lactic acid and antioxidants and stored under modified
atmospheres. Food Microbiol. 20, 17.
DJENANE, D., LEFSIH, K., YANGELA, Y. and RONCALS, P.
2011a. Composition chimique et activit anti-Salmonella
Enteritidis CECT 4300des huiles essentielles dEucalyptus
globulus, Lavandula angustifolia et Satureja hortensis; Tests in
vitro et efficacit sur les ufs entiers liquides conservs
7 1C. Phytothrapie (accepted for publication).
DJENANE, D., YANGELA, J., AMROUCHE, T., BOUBRIT, S.,
BOUSAD, N. and RONCALS, P. 2011b. Chemical
composition and antimicrobial effects of essential oils of
Eucalyptus globulus, Myrtus communis and Satureja hortensis
against Escherichia coli O157:H7 and Staphylococcus aureus in
minced beef. Food Sci. Technol. Int. (in press). DOI:
10.1177/1082013211398803.
DJENANE, D., YANGELA, J., MONTAS, L., DJERBAL, M.
and RONCALS, P. 2011c. Antimicrobial activity of Pistacia
lentiscus and Satureja montana essential oils against Listeria
monocytogenes CECT 935 using laboratory media; efficacy and
synergistic potential in minced beef. Food Control 22,
10461053.
DORMAN, H.J. and DEANS, S.G. 2000. Antimicrobial agents
from plants: Antibacterial activity of plant volatile oils. J. Appl.
Microbiol. 88, 308316.
European Food Safety Authority 2009. The community summary
report on trends and sources of zoonoses and zoonotic agents
in the European Union in 2007. EFSA J. 223, 4312.

45

ANTIMICROBIALS FOR C. JEJUNI IN CHICKEN

FISHER, K. and PHILLIPS, C.A. 2006. The effect of lemon, orange

and bergamot essential oils and their components on the
survival of Campylobacter jejuni, Escherichia coli O157, Listeria
monocytogenes, Bacillus cereus and Staphylococcus aureus in
vitro and in food systems. J. Appl. Microbiol. 101,
12321240.
FITZGERALD, D.J., STRATFORD, M. and NARBAD, A. 2003.
Analysis of the inhibition of food spoilage yeasts by vanillin.
Int. J. Food Microbiol. 86, 113122.
GILL, A.O., DELAQUIS, P., RUSSO, P. and HOLLEY, R.A. 2002.
Evaluation of antilisterial action of cilantro oil on vacuum
packed ham. Int. J. Food Microbiol. 73, 8392.
HOLLEY, R.A. and PATEL, D. 2005. Improvement in shelf-life
and safety of perishable foods by plant essential oils and smoke
antimicrobials. Food Microbiol. 22, 273292.
KOUTSOUDAKI, C., KRSEK, M. and RODGER, A. 2005.
Chemical composition and antibacterial activity of the essential
oil and the gum of Pistacia lentiscus var. chia. J. Agric. Food
Chem. 53, 76817685.
LEE, C.F., HAN, C.K. and TSAU, J.L. 2004. In vitro inhibitory
activity of Chinese leek extract against Campylobacter species.
Int. J. Food Microbiol. 94, 169174.
LIRA, P.D., LEO RETTA, D., TKACIK, E., RINGUELET, J.,
COUSSIOA, J.D., VAN BAREN, C. and BANDONI, A.L. 2009.
Essential oil and by-products of distillation of bay leaves
(Laurus nobilis L.) from Argentina. Ind. Crop. Prod. 30,
259264.
LIS-BALCHIN, M. and DEANS, S.G. 1997. Bioactivity of selected
plant essential oils against Listeria monocytogenes. J. Appl.
Microbiol. 82, 759762.
MARINO, M., BERSANI, C. and COMI, G. 1999. Antimicrobial
activity of the essential oils of Thymus vulgaris L. measured
using a bioimpedometric method. J. Food Prot. 62, 10171023.
MARINO, M., BERSANI, C. and COMI, G. 2001. Impedance
measurements to study the antimicrobial activity of essential
oils from Lamiaceae, Compositae. Int. J. Food Microbiol. 67,
187195.
MARTNEZ, L., DJENANE, D., CILLA, I., BELTRN, J.A.
and RONCALS, P. 2006. Antioxidant effect of rosemary,
borage, green tea, pu-erh tea and ascorbic acid on fresh pork
sausages packaged in modified atmosphere. Influence
of the presence of sodium chloride. J. Sci. Food Agric. 86,
12981307.
MASTELIC, J. and JERKOVIC, I. 2003. Gas
chromatography-mass spectrometry analysis of the free and
glycoconjugated aroma compounds of the seasonally collected
Satureja montana L. Food Chem. 80, 135140.
MOUREY, A. and CANILLAC, N. 2002. Anti-Listeria
monocytogenes activity of essential oils components of conifers.
Food Control 13, 289292.
NANNAPANENI, R., CHALOVA, V.I., CRANDALL, P.G., RICKE,
S.C., JOHNSON, M.G. and OBRYAN, C.A. 2009.
Campylobacter and Arcobacter species sensitivity to
commercial orange oil fractions. Int. J. Food Microbiol. 129,
4349.

46

D. DJENANE ET AL.

NAUTA, M., HILL, A., ROSENQUIST, H., BRYNESTAD, S.,

FETSCH, A., VAN DER LOGT, P., FAZIL, A., CHRISTENSEN,
B., KATSMA, E., BORCK, B. et al. 2009. Review: A comparison
of risk assessments on Campylobacter in broiler meat. Int. J.
Food Microbiol. 129, 107123.
OKE, F., ASLIM, B., OZTURK, S. and ALTUNDAG, S. 2009.
Essential oil composition, antimicrobial and antioxidant
activities of Satureja cuneifolia Ten. Food Chem. 112,
874879.
OUSSALAH, M., CAILLET, S., SAUCIER, L. and LA CROIX, M.
2007. Inhibitory effects of selected plant essential oils on the
growth of four pathogenic bacteria: E. coli O157:H7,
Salmonella Typhimurium, Staphylococcus aureus and Listeria
monocytogenes. Food Control 18, 414420.
PATTNAIK, S., SUBRAMANYAM, V.R., BAPAJI, M. and KOLE,
C.R. 1997. Antibacterial and antifungal activity of aromatic
constituents of essential oils. Microbios 89, 3946.
PFALZGRAF, A., FRIGG, M. and STEINHART, H. 1995. Alpha
tocopherol contents and lipid oxidation in pork muscle and
adipose tissue during storage. J. Agric. Food Chem. 43,
13391342.
PONCE, A.G., FRITZ, R., DEL VALLE, C. and ROURA, S.I. 2003.
Antimicrobial activity of essential oils on the native microflora
of organic Swiss chard. Lebens. Wiss. Technol. 36, 679684.
REICH, F., ATANASSOVA, V., HAUNHORST, E. and KLEIN, G.
2008. The effects of Campylobacter numbers in caeca on the
contamination of broiler carcasses with Campylobacter. Int. J.
Food Microbiol. 127, 116120.
SNCHEZ-ESCALANTE, A., DJENANE, D., BELTRN, J.A. and
RONCALS, P. 2003. Antioxidant action of borage, rosemary,
oregano and ascorbic acid in beef patties packaged in modified
atmosphere. J. Food Sci. 68, 339344.
SHARIFIFAR, F., MOSHAFI, M.H., MANSOURI, S.H.,
KHODASHENAS, M. and KHOSHNOODI, M. 2007. In vitro
evaluation of antibacterial and antioxidant activities of the
essential oil and methanol extract of endemic Zataria
multiflora Boiss. Food Control 18, 800805.
SLAVKOVSKA, V., JANEIC, R., BOJOVIC, S., MILOSAVLJEVIC,
S. and DJOKOVIC, D. 2001. Variability of essential oils of
Satureja montana L. and Satureja kitaibelii Wierzb. Ex Heuff.
from the central part of the Balkan peninsula. Phytochemistry
57, 7176.
SMITH-PALMER, A., STEWART, J. and FYFE, L. 2001. The
potential application of plant essential oils as natural food
preservatives in soft cheese. Food Microbiol. 18, 463470.
SOLOMAKOS, N., GOVARIS, A., KOIDIS, P. and BOTSOGLOU,
N. 2008. The antimicrobial effect of thyme essential oil, nisin,
and their combination against Listeria monocytogenes in
minced beef during refrigerated storage. Food Microbiol. 25,
120127.
SON, I., ENGLEN, M.D., BERRANG, M.E., FEDORKA-CRAY, P.J.
and HARRISON, M.A. 2007. Antimicrobial resistance of
Arcobacter and Campylobacter from broiler carcasses. Int. J.
Antimicrob. Agents 29, 451455.
SPSS 1995. SPSS for Windows, 6.1.2, SPSS Inc, Chicago, IL.

Journal of Food Safety 32 (2012) 3747 2011 Wiley Periodicals, Inc.

D. DJENANE ET AL.

SUZUKI, H. and YAMAMOTO, S. 2009. Review: Campylobacter

contamination in retail poultry meats and by-products
in Japan: A literature survey. Food Control 20, 531
537.
TEPE, B., SIHOGLU-TEPE, A., DAFERERA, D., POLISSIOU, M.
and SOKMEN, A. 2007. Chemical composition and
antioxidant activity of the essential oil of Clinopodium vulgare
L. Food Chem. 103, 766770.
WEGENER, H.C. 2010. Danish initiatives to improve the safety of
meat products. Meat Sci. 84, 276283.

Journal of Food Safety 32 (2012) 3747 2011 Wiley Periodicals, Inc.

ANTIMICROBIALS FOR C. JEJUNI IN CHICKEN

ZHAO, J., LI, Y., LIU, Q. and GAO, K. 2010. Antimicrobial

activities of some thymol derivatives from the roots of Inula
hupehensis. Food Chem. 120, 512516.
ZILBAUER, M., NICK DORRELL, N., WREN, B.W. and
BAJAJ-ELLIOTT, M. 2008. Campylobacter jejuni-mediated
disease pathogenesis: An update. Trans. R. Soc. Trop. Med. Hyg.
102, 123129.
ZRIRA, S., ELAMRANI, A. and BENJILALI, B. 2003. Chemical
composition of the essential oil of Pistacia lentiscus L. from
Morocco a seasonal variation. Flav. Frag. J. 18, 475480.

47