International Journal of Food Microbiology 37 (1997) 155162

Antibacterial activity of selected fatty acids and essential oils

against six meat spoilage organisms
a, b c a
Blaise Ouattara *, Ronald E. Simard , Richard A. Holley , Gabriel J.-P. Piette ,
a
Andre Begin

a
Agriculture and Agri-Food Canada, Food Research and Development Centre, 3600 Casavant Blvd. West, St. Hyacinthe, Quebec ,
Canada, J2 S 8 E3
b

Departement de Science et Technologie des Aliments, Faculte des Sciences de l Agriculture et de l Alimentation, Universite Laval,
, Canada, G1 K 7 P4
Quebec
c
Department of Food Science, Faculty of Agricultural and Food Sciences, University of Mannitoba, Winnipeg, Mannitoba, Canada,
R3 T 2 N2

Received 2 December 1996; received in revised form 16 May 1997; accepted 10 June 1997

Abstract

The antibacterial activity of selected fatty acids and essential oils was examined against two gram-negative (Pseudomonas
fluorescens and Serratia liquefaciens), and four gram-positive (Brochothrix thermosphacta, Carnobacterium piscicola,
Lactobacillus curvatus, and Lactobacillus sake) bacteria involved in meat spoilage. Various amounts of each preservative
were added to brain heart infusion or MRS (deMan, Rogosa and Sharpe) agars, and the minimum inhibitory concentration
was determined for each organism. Essential oils were analysed by gasliquid chromatography to determine the
concentration of selected components commonly found in spices. B. thermosphacta, P. fluorescens and S. liquefaciens were
not affected by fatty acids, and generally overcame the inhibitory effect of essential oils after 24 h of exposure. Among the
fatty acids, lauric and palmitoleic acids exhibited the greatest inhibitory effect with minimum inhibitory concentrations of
250 to 500 mg / ml, while myristic, palmitic, stearic and oleic acids were completely ineffective. For essential oils, clove,
cinnamon, pimento, and rosemary were found to be the most active. The 1 / 100 dilution of those oils inhibited at least five of
the six tested organisms. A relationship was found between the inhibitory effect of essential oils and the presence of eugenol
and cinnamaldehyde. 1997 Elsevier Science B.V.

Keywords: Fatty acids; Essential oils; Meat; Spoilage; Bacteria

1. Introduction products require longer shelf-life and greater assur-

The problem of safe preservation in the meat
industry has grown to be more complex as todays
*Corresponding author. Tel: 514 773 1105; Fax: 514 773 8461.
ance of protection from microbial spoilage. Many
attempts have been made to control microbial growth
at the surface of meat and meat products with
antimicrobial chemicals. For example, significant
reductions of microbial growth were obtained by

0168-1605 / 97 / $17.00 1997 Elsevier Science B.V. All rights reserved.

PII S0168-1605( 97 )00070-6
156 B. Ouattara et al. / International Journal of Food Microbiology 37 (1997) 155 162
dipping or spraying meat with organic acid solutions
(Abugroun et al., 1993; Anderson and Marshall,
1989). However, preservatives could not be stabi-
lized at the surface of food due to evaporation,
neutralization (Siragusa and Dickson, 1992), and
diffusion into the matrix (Torres et al., 1985).
Fatty acids and essential oils have also been
shown to possess antibacterial and antifungal ac-
tivities against many plant and food microorganisms
(Kabara, 1981; Shelef et al., 1980; Russel, 1991).
Gram-negative bacteria were shown to be generally
more resistant than gram-positive ones to the antago-
nistic effects of fatty acids and essential oils because
of their cell wall lipopolysaccharide (Kabara, 1979;
Branen et al., 1980; Russel, 1991) but this was not
always true (Karapinar and Aktug, 1987). In addi-
tion, most studies to date have been done with
pathogens such as Salmonella typhimurium and
Staphylococcus aureus (Karapinar and Aktug, 1987;
Paster et al., 1990; Juven et al., 1994), Listeria
monocytogenes (Aureli et al., 1992; Wang and John-
son, 1992), Vibrio parahaemoliticus (Karapinar and
Aktug, 1987; Shelef et al., 1980), and Clostridium
botulinum (Ababouch et al., 1992), and little is
known about the effect of these compounds on meat
spoilage bacteria such as Carnobacterium piscicola,
Lactobacillus curvatus and Lactobacillus sake.
An investigation is currently under way in our
laboratory to develop active packaging materials for
the preservation of meat products. As a first step, it
was necessary to know how the regular meat flora
was affected by antibacterial agents currently ap-
proved for food use, in particular fatty acids and
essential oils. The purpose of the present study was
therefore to evaluate the efficacy of various fatty
acids and essential oils to control the growth of meat
spoilage organisms.
2. Materials and methods
2.1. Organisms and cultures
The following organisms were obtained from the
American Type Culture Collection (Rockville, MD,
USA); Carnobacterium piscicola (ATCC 43224),
Lactobacillus curvatus (ATCC 25601), and Lac-
tobacillus sake (ATCC 15521). Pseudomonas
fluorescens and Brochothrix thermosphacta were
isolated from beef stored at 48C (Farber and Idziak,
1984). Serratia liquefaciens was isolated from vac-
uum packaged bologna (Food Research and De-
velopment Centre, St. Hyacinthe, Quebec).
P. fluorescens, B. thermosphacta, and S. liquefa-
ciens were first inoculated and grown aerobically on
brain heart infusion agar (BHI, Difco Laboratories,
Detroit, MI, USA). C. piscicola, L. curvatus, and L.
sake were similarly inoculated and grown on lac-
tobacilli MRS agar (Difco), in an atmosphere en-
riched in hydrogen and carbon dioxide (Gaspak
Anaerobic System; Becton Dickinson, Cockeysville,
MD, USA). All incubations were done at 208C.
Bacterial cells were subsequently harvested and
resuspended in reconstituted skim milk (skim milk
powder in deionized water, 20% w / v final con-
centration), containing 5% sucrose (w / v), and
lyophilized to obtain stock cultures.
To prepare working cultures, stock cultures were
standardized through two successive 24 h growth
cycles in the appropriate broth (BHI or MRS)
without agitation. Cells from the standardized cul-
tures were then inoculated in fresh medium and
incubated (208C without agitation) for 6 h (C.
piscicola for 9 h) to obtain working cultures con-
taining approximately 10 7 colony forming units
(CFU) / ml.
2.2. Preparation of the antibacterial media
Analytical grade free fatty acids [lauric (C 12:0 ),
myristic (C 14:0 ), palmitic (C 16:0 ), palmitoleic (C 16:1 ),
stearic (C 18:0 ), oleic (C 18:1 ), linoleic (C 18:2 ), and
linolenic (C 18:3 )], with a purity $ 98% were ob-
tained from Sigma (St. Louis, MO, USA). The acids
were first dissolved in 95% ethanol (Ababouch et al.,
1992), and the solutions were added to 250 ml
bottles of sterile BHI or MRS molten agar in
concentrations ranging from 100 to 2500 mg / ml, in
increments of 50 mg / ml from 100 to 500 mg / ml, and
of 100 mg / ml from 500 to 1000 mg / ml. The
contents of each bottle were then dispensed into
sterile Petri plates and left to solidify. The maximum
concentration of ethanol in the agar was 2.5% (v / v),
which was shown in preliminary trials to have no
inhibitory effect upon the microorganisms used in
this study.
Eight essential oils (cinnamon, ELB 40404; clove,
ELB 41312; cumin, ELB 41402; garlic, EB 40892;
 B. Ouattara et al. / International Journal of Food Microbiology 37 (1997) 155 162 157

oregano, ELB 41401; black pepper, EB 33423; limonene, and g-terpinene were obtained from Fluka
pimento, ELB 41441; thyme, ELB 41403) were ChemikaBiochemika (Buchs, Switzerland). Cam-
provided by Food Ingredients (Mississauga, Ontario, phene, linalool, a-terpinene, and g-terpinene were of
Canada). Rosemary oil, 8136-L was obtained from technical quality (9095% purity) while all the
Kalsec (Kalamazoo, MI, USA). Oils were manually others compounds were at least 97% pure.
mixed with sterile molten BHI or MRS agar main- A Hewlett-Packard model 5890 gas chromato-
tained at 458C, to dilutions of 1 / 10, 1 / 100, and graph equipped with a 1-mm DB-1 fused-silica
1 / 1000. The molten agars containing essential oils column 30 m 3 0.316 mm (J and W Scientific,
were poured into sterile Petri plates and left to Folsom, CA, USA) was used to determine the
solidify. concentration of the seventeen substances in the
selected essential oils. The split injector was set at
2.3. Growth inhibition experiments ratio of 18:1, and the carrier gas (He) flow at 1.0
ml / min. The oven temperature was programmed to
Petri plates of BHI or MRS agar containing rise 2 C8 / min from 908C to 1158C, 5 C8 / min from
various concentrations of fatty acids or essential oils 1158C to 2008C, and remained isothermal at the final
were inoculated with the selected organisms. The temperature (2008C) for 4 min. Samples of essential
working cultures were diluted (1 / 100) in peptone oils injected in the gas chromatograph consisted of 1
water, and 0.1 ml of the diluted cultures was spread ml of 250 mg / ml (rosemary), and 50 mg / ml (other
on the surface of the solidified agar plates. The essential oils) solutions in ethyl acetate.
positive controls for growth consisted of BHI and
MRS agar without preservative, inoculated with the
diluted working cultures. Uninoculated plates con- 3. Results
taining either fatty acids or essential oils, served as
negative controls. Test and control plates were then 3.1. Fatty acids
incubated at 208C under aerobic conditions for B.
thermosphacta, P. fluorescens, and S. liquefaciens, or All the fatty acids failed to inhibit B. thermos-
in H 2 - and CO 2 -enriched atmosphere for C. pis- phacta, P. fluorescens, and S. liquefaciens at con-
cicola, L. curvatus, and L. sake. centrations up to 2500 mg / ml (results not shown).
Three Petri plates were used to test the inhibitory The inhibitory effects against the three other bacteria
effect for each organism and each level of each (C. piscicola, L. curvatus, and L. sake) are presented
preservative, and the experiment was performed in Table 1. All were unaffected by myristic, palmitic,
twice. Plates were checked for presence or absence stearic, and oleic acids at the concentrations tested.
of colonies after incubation for 24 and 48 h. The Lauric, palmitoleic, linoleic, and linolenic acids
absence of colonies on all the three plates of a exhibited various inhibitory activity with lauric and
treatment was considered as an inhibitory effect. The palmitoleic acids having the greatest effect. Among
lowest concentration of fatty acids or essential oils
required to inhibit the growth of the test micro-
organisms was designated as the minimum inhibitory Table 1
Minimum inhibitory concentration (mg / ml) of fatty acids against
concentration. meat spoilage bacteria
Fatty acids C. piscicola L. curvatus L. sake
2.4. Analysis of essential oils
Lauric C 12:0 250 500 500
Seventeen substances commonly found in spices Myristic C 14:0 NI a NI NI
Palmitic C 16:0 NI NI NI
were used for this experiment: allylsulfide, carvacrol, Palmitoleic C 16:1 350 450 450
camphor, cineole, eugenol, 4-iso-propylben- Stearic C 18:0 NI NI NI
zaldehyde, trans-cinnamaldehyde, myrcene, a-ter- Oleic C 18:1 NI NI NI
pineol, and thymol were purchased from Aldrich Linoleic C 18:2 600 650 650
Chemical (Milwaukee, WI, USA); geraniol, linalool, Linolenic C 18:3 500 500 650
a
and a-terpinene from Sigma; camphene, carvon, NI: No inhibition at concentrations up to 2500 mg / ml.
158 B. Ouattara et al. / International Journal of Food Microbiology 37 (1997) 155 162

the organisms which were affected by fatty acids, C.
piscicola was the most susceptible.
3.2. Essential oils
All the essential oils tested for antibacterial activi-
ty were ineffective at the 1 / 1000 dilution (results not
shown). The inhibitory properties observed with the
1 / 100 and 1 / 10 dilutions are shown in Table 2. The
strongest effects were obtained with clove, cin-
namon, pimento, and rosemary oils, for which the
1 / 100 dilution inhibited at least five of the six tested
organisms. However, pimento oil was not able to
maintain the inhibitory effect over 24 h. All the other
oils were weakly active.
Gram-positive and gram-negative bacteria were
generally affected in the same manner within 24 h of
exposure, but extension of the inhibitory effects up
to 48 h was less often observed with the gram-
negative bacteria (Table 2). For example, P. fluores-
cens and S. liquefaciens, which were affected by the
1 / 100 dilution of cinnamon, clove, and rosemary
were no longer inhibited after 48 h, except for S.
liquefaciens in the presence of clove oil. In contrast,
three of the four gram-positive organisms (C. pis-
cicola, L. curvatus, and L. sake) continued to be
inhibited by the same oils at that dilution. Of the
gram-positive bacteria, B. thermosphacta exhibited
resistance similar to those of the two gram-negative
bacteria tested.
The contents of the essential oils in the seventeen
selected substances is shown in Table 3. In general,
the sum of selected substances which were identified
and quantified constituted a small proportion of the
total mass of each essential oil, with values ranging
from 0.15% for rosemary oil to 19.94% for clove oil.
In addition, three of the four most active essential
oils (which inhibited more than five organisms at the
1 / 1000 dilution) contained a significant amount of
eugenol: clove (19.81%); pimento (9.33%); and
cinnamon (5.38%). Cinnamon oil also contained
large amounts of cinnamaldehyde (5.37%). The least
most active oil (rosemary oil) contained camphor as
its major component, but in a low amount (0.10%).
Among the less active essential oils (those which
inhibited only two or less than two organisms at the
1 / 1000 dilution), only thyme oil contained eugenol
and cinnamaldehyde, but these were present in small
amounts (0.01% for each of the two components).
On the other hand, greater concentrations of other
components were found in those oils: carvacrol in
oregano oil (5.19%); a-terpinene in cumin and
thyme oils (1.15% and 1.64%, respectively); and
thymol in thyme oil (2.40%).
4. Discussion
The two gram-negative bacteria (P. fluorescens,
and S. liquefaciens) were unaffected by fatty acids at
concentrations up to 2500 mg / ml. This was to be
expected since several other studies (Kabara, 1979,
1981; McKellar et al., 1992) reported that gram-
negative bacteria were resistant to the inhibitory
effects of medium and long chain fatty acids and
their derivatives. This resistance has been attributed
to the presence of cell wall lipopolysaccharides,
which can screen out the fatty acids; the lipids are

Table 2
Inhibitory properties of diluted essential oils toward meat spoilage bacteria a
Essential oils B. thermosphacta P. fluorescens S. liquefaciens C. piscicola L. curvatus L. sake
b
Cinnamon 11 11 11 (1 1) (1 1) (1 1)
Clove (1 1) 11 (1 1) (1 1) (1 1) (1 1)
Cumin 1c 1 1 1 1 1
Garlic 11 1 11 1 1 1
Oregano 1 1 1 1 1 1
Black pepper 1 1 1 1 11 11
Pimento 1 11 11 11 11 11
Rosemary 11 11 11 (1 1) (1 1) (1 1)
a
Bacteria were tested at 10 5 CFU / ml, at both 24 and 48 h.
b
1 1 : Inhibition by 1 / 100 dilution of essential oils.
c
1 : Inhibition by 1 / 10 dilution of essential oils.
() Inhibition extended to 48 h by 1 / 100 dilution of essential oils.
 B. Ouattara et al. / International Journal of Food Microbiology 37 (1997) 155 162 159

Table 3
Quantitative determination of selected authentic antibacterial components in essential oils a
Most efficient oils Least efficient oils
Compound Clove Cinnamon Pimento Rosemary Garlic Black pepper Oregano Cumin Thyme
Total identified 19.94 11.20 9.83 0.15 0.26 2.94 6.49 5.50 6.78
Allylsulfide b 0.22
Camphene 0.02 - 0.02 0.05 0.09
Camphor 0.10 0.09 0.10
Carvacrol 0.03 0.01 5.19 0.04 0.31
Carvon 0.07 0.08 0.01
Cinnamaldehyde 5.37 0.01
Eugenol 19.81 5.38 9.33 0.01
Fenchon 0.02 0.04 0.02 0.03 0.02
Geraniol
4-Isopropylbenzaldehyde 0.05 0.01 0.02 1.95
Limonenecineole 0.05 0.11 1.12 0.08 1.24 1.31
Linalool 0.02 0.15 0.11 0.12 0.14 0.07 0.26
Myrcene 0.03 0.03 0.01 0.93 0.05 0.55 0.49
a-Terpinene 0.06 0.06 0.04 0.29 1.15 1.64
g-Terpinene 0.06 0.05 0.49 0.11 0.04 0.08
a-Terpineol 0.03 0.04 0.04 0.02 0.02 0.08 0.07 0.43 0.05
Thymol 0.09 0.03 0.01 0.37 2.40
a
The results are given in % of the total mass of oils.
b
Component not present in the oil.

thus prevented from accumulating on the transport-
ing cell membrane, and from entering into the cells
(Kabara, 1979; Branen et al., 1980; Russel, 1991).
B. thermosphacta, a gram-positive bacterium, also
exhibited resistance to fatty acids, but little infor-
mation is available about its sensitivity to challenge
by fatty acids. Macaskie (1982) reported that growth
rates and numbers of B. thermosphacta were both
reduced in the presence of 0.5 mmol / l of palmitic
acid. However, both the determination of palmitic
acid uptake and the determination of the inhibition of
substrate uptake by palmitic acid failed to explain the
mechanism by which B. thermosphacta was in-
hibited. Similar resistance of gram-positive organ-
isms was reported by Tsuchido et al. (1993) working
with Bacillus subtilis. They found mutants which
were tolerant to the lytic action of sucrose esters of
long-chain fatty acids.
Among the saturated fatty acids under study,
lauric acid exhibited the greatest inhibitory effect
against C. piscicola, L. curvatus, and L. sake while
all the other saturated fatty acids with chain length
between C 14 and C 18 were completely ineffective.
These results are consistent with previous reports
about the antibacterial activity of saturated fatty
acids with lauric acid being the most effective
(Kabara, 1979; Branen et al., 1980; Babic et al.,
1994). For saturated fatty acids, hydrophobic groups
have been shown to have the greatest influence on
antibacterial activity (Branen et al., 1980), but
increasing hydrophobicity with longer chain length
may reduce their solubility in aqueous systems. Thus
hydrophobic groups may be prevented from reaching
sufficient concentration to interact with hydrophobic
proteins or lipids on the bacterial cell surface (Wang
and Johnson, 1992). Lauric acid has been reported to
have the best balance between hydrophobic and
hydrophilic groups (Branen et al., 1980; Kabara et
al., 1977).
It is known that unsaturated fatty acids with chain
lengths of C 14 or longer are more active against
microorganisms than the corresponding saturated
fatty acids (Kabara, 1981). Also, the inhibitory
effects of unsaturated fatty acids are increased as the
number of double bonds in the molecule increases
(Kabara, 1979). In agreement with that observation,
palmitoleic acid was found to be more active than
myristic and palmitic acids (this study), and the
antibacterial efficacies of C 18 unsaturated fatty acids
were in the following order: linolenic (C 18:3 ) .
linoleic (C 18:2 ) . oleic (C 18:1 ). Similar results have
been reported by Wang and Johnson (1992) who
160 B. Ouattara et al. / International Journal of Food Microbiology 37 (1997) 155 162
found that linolenic acid was more effective against
Listeria monocytogenes than linoleic and oleic acids.
The fact that palmitoleic acid and C 18 unsaturated
fatty acids are active in spite of their long carbon
chain suggests that the hydrophobic / hydrophilic
balance alone cannot explain the observed inhibitory
effects. This activity may be related to other factors
such as a peroxidative process involving hydrogen
peroxide and bacterial iron as reported by Wang and
Johnson (1992).
In the study on the antibacterial activity of essen-
tial oils, no obvious difference in susceptibility was
found between gram-negative and gram-positive
bacteria after 24 h of exposure to essential oils. Data,
however, showed that the extent of the inhibitory
effect up to 48 h was mostly observed with gram-
positive organisms. This is supported by many other
reports on the greater susceptibility of gram-positive
bacteria to the inhibitory effect of essential oils and
their components (Shelef et al., 1980; Farag et al.,
1989; Chanegriha et al., 1994). As reported for fatty
acids, the cell wall lipopolysaccharides of gram-
negative bacteria may prevent active components
from reaching the cytoplasmic membrane.
However, the greater resistance of gram-negative
bacteria may not be an overall trend since B.
thermosphacta (gram-positive) was as resistant as S.
liquefaciens (gram-negative). Similar results have
been reported by Kim et al. (1995b) who found that
L. monocytogenes (gram-positive) was more resistant
to the inhibitory effects of eleven essential oil
constituents than the gram-negative bacteria tested
under the same conditions, including Escherichia
coli, E. coli O157:H7, Salmonella typhimurium and
Vibrio vulnificus. It seems that the variability of the
resistance of gram-positive bacteria to the inhibitory
effect of essential oils may be due to differences
between strains of the same bacterial species. This
hypothesis was recently confirmed by Sivropoulou et
al. (1996) with two strains of Staphylococcus aureus
in the presence of carvacrol and thymol.
(1988) reviewed the literature reporting the
Zaka
antimicrobial activity of many spices and classified
their activities as strong, medium, or weak. Accord-
ing to this ranking, several studies (Conner, 1993;
Aureli et al., 1992; Shelef et al., 1980) showed that
cinnamon, clove, pimento, thyme, oregano, and
rosemary had strong and consistent inhibitory effects
against various pathogens and spoilage bacteria. In
agreement with this finding, four of the essential oils
tested in this study (cinnamon, clove, pimento, and
rosemary) exhibited a strong inhibitory effect toward
selected meat spoilage bacteria. The antibacterial
activities have been attributed to the presence of
some volatile constituents in the oils. Bullerman et
al. (1977) found that cinnamon and clove contained
cinnamaldehyde and eugenol as major constituents
which represented 6575% and 9395% of the total
volatile oils, respectively, and which were respon-
sible for the antibacterial effect. In oregano and
thyme, the major antibacterial constituents have been
identified as carvacrol (6279%), and thymol (42%)
respectively (Farag et al., 1989; Sivropoulou et al.,
1996).
The means by which microorganisms are inhibited
by essential oils seems to involve different modes of
action. The most frequent inhibitions involve phen-
olic components of oils which sensitize the phos-
pholipid bilayer of the cell membrane, causing an
increase of permeability and leakage of vital intracel-
lular constituents (Kim et al., 1995b; Juven et al.,
1994), or impairment of bacterial enzyme systems
(Wendakoon and Sakaguchi, 1995; Farag et al.,
1989). A number of reports indicated that essential
oils containing carvacrol, eugenol, or thymol had the
highest antibacterial performances (Kim et al.,
1995b; Lattaoui and Tantaoui-Elaraki, 1994; Suresh
et al., 1992). For example, Suresh et al. (1992) found
that eugenol was more bactericidal against Es-
cherichia coli, Enterobacter sakazakii, and Klebsiel-
la pneumoniae than several antibiotics including
ampicillin, erythromycin, and sulphamethizole.
Among non-phenolic compounds of essential oils,
cinnamaldehyde has been shown to possess anti-
bacterial properties by inhibiting amino acid de-
carboxylase activity (Didry et al., 1993; Wendakoon
and Sakaguchi, 1995). Baranowski and Nagel (1982)
reported that allylhydroxycinnamates, which are
quite similar to cinnamaldehyde inhibited P. fluores-
cens by a specific mode of action related to cellular
energy depletion.
The antibacterial activity of eugenol and cinnamal-
dehyde was supported by the results obtained by the
gasliquid chromatographic analysis of the essential
oils, although components quantified constituted only
a small proportion of the oils. Cinnamon and clove
oils which were among the most active oils con-
tained the largest amounts of eugenol and cinnamal-
 B. Ouattara et al. / International Journal of Food Microbiology 37 (1997) 155 162
dehyde. Also eugenol and cinnamaldehyde were
slightly or not present in the oils which produced
small inhibitory effects (inhibition of two or less
than two organism at 1 / 1000 dilution). Therefore,
the presence of eugenol or cinnamaldehyde was
directly related to the antibacterial properties of
tested essential oils.
Our results, however, failed to confirm the inhib-
itory effect of oregano and thyme although those oils
contained high concentrations of phenolic com-
pounds (carvacrol and thymol respectively). Juven et
al. (1994) reported that in presence of a high oxygen
tension, thyme and oregano oils may be inactivated
by oxidation of their phenolic components. In the
present study, their effectiveness was not enhanced
when the test was done under anaerobic conditions
(inhibition test against C. piscicola, L. curvatus, and
L. sake). Therefore, the low antibacterial activity
reported here for thyme and oregano oils could not
be explained in terms of the oxygen tension hypoth-
esis. It is most likely that the weak efficacy of
carvacrol and thymol-containing oils (oregano and
thyme) found in the present study may be due to
some other factors such as insolubility in aqueous
media (Juven et al., 1994), pH of the medium
(Thompson, 1990), or seasonal and intraspecific
variation of essential oil composition (McGimpsey et
al., 1994; Kokkini and Vokou, 1989; Sivropoulou et
al., 1996). For example, an essential oil from
Origanum vulgare has been reported by Sivropoulou
et al. (1996) to eliminate S. aureus at dilutions up to
1 / 10000, but the oil sample used contained carvacrol
at a concentration of 79.58% compared to 5.19% in
the commercial oregano oil tested in the present
study.
In the present study, rosemary oil was as inhib-
itory as cinnamon and clove oils. Yet, rosemary did
not contain cinnamaldehyde nor eugenol, and all the
other background components screened were present
in small amounts. Camphor (0.10%) was the only
component which was present in rosemary oil in
concentrations higher than in the other oils under
study. Therefore, the antibacterial efficacy of rosem-
ary oil could be at least partly related to the presence
of camphor. This is supported by the report of
Lattaoui and Tantaoui-Elaraki (1994) who found that
in some essences, minor compounds could have a
huge antibacterial impact. Also, the small amount of
the total identified components (0.15%) in rosemary
161
oils suggests that some other components may have
contributed to its high antibacterial action.
The present study on the inhibitory effects of fatty
acids and essential oils on meat spoilage bacteria was
done under specific, controlled conditions (BHI and
MRS agars). Even though some of these compounds
showed consistent antibacterial activities against
meat spoilage bacteria, the extrapolation of these
results to meat systems must be done with caution.
Bacteria present on meat surfaces may attach firmly
resulting in reduced exposure to essential oils or fatty
acids. Proteins and lipid components of meat can
also interact with the active components of anti-
bacterial compounds as reported by Kim et al.
(1995a). Also, for subsequent use as components of
active packages, additional experiments must be
done to determine the ease with which fatty acids
and essential oils can be incorporated into packaging
films and their diffusion rates from the surface of the
product to the interior must be characterized.
Acknowledgments
This research was made possible by the financial
support of Programme Canadien des Bourses de la
Francophonie, Agence Canadienne du Developpe-
ment International, Ottawa, Ontario. The expert
technical assistance of Yves Raymond is greatly
appreciated
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