Food Control 34 (2013) 619e623

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Food Control
journal homepage: www.elsevier.com/locate/foodcont

Synergistic interactions of cinnamaldehyde in combination with

carvacrol against food-borne bacteria
Haiqing Ye a, Suxia Shen a, Jingyue Xu a, Songyi Lin b, *, Yuan Yuan a, Gregory S. Jones c
a
Department of Food Quality and Safety, Jilin University, Changchun 130062, PR China
b
Laboratory of Nutrition and Functional Food, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China
c
Department of Food, Nutrition and Packaging Sciences, Clemson University, Clemson, SC 29634, USA

a r t i c l e i n f o a b s t r a c t

Article history: In order to select effective and safe natural antibacterial ingredients, more than 30 kinds of plant extracts
Received 17 March 2013 were selected for their suitability as antibacterial agents. A standard broth microdilution method was
Received in revised form used to evaluate their antimicrobial activity, a cytotoxicity test was used to detect their safety, and a
21 May 2013
synergy assay was used to determine which combinations have a synergistic effect. Then time-kill curves
Accepted 28 May 2013
were used to further verify their bactericidal capacity. As a result of these tests, cinnamaldehyde and
carvacrol were identified first, then subsequently verified to be the most effective and safe natural active
Keywords:
substances. It was found that all of the tested bacteria strains were sensitive to cinnamaldehyde and
Cinnamaldehyde
Carvacrol
carvacrol. The combination of cinnamaldehyde with carvacrol also showed good synergistic antibacterial
Antibacterial activity effect against 7 of the 11 tested bacterial strains. The time-kill assay verified synergism for the cinna-
Synergy maldehyde/carvacrol combination toward Escherichia coli and Staphylococcus aureus. These results
Time-kill assay indicated that the combination of cinnamaldehyde and carvacrol may serve as a promising naturally
sourced food preservative with excellent bactericidal activity against common food spoilage
microorganisms.
Ó 2013 Elsevier Ltd. All rights reserved.

1. Introduction
Food spoilage is a major problem that restricts the storage and
transportation of food and also seriously impacts food safety. It
drives us to find better ways to extend the shelf-life of foods. There
are a lot of strategies to prevent pathogenic and spoilage microor-
ganisms in food, and using chemical preservative is one of the
important ways (Sanla-Ead, Jangchud, Chonhenchob, & Suppakul,
2012). However, there is currently a strong debate about the
safety aspects of chemical preservatives, because they are consid-
ered to have many carcinogenic and teratogenic attributes as well
as residual toxicity, consumers tend to be suspicious of chemical
additives. For these reasons, alternative means are required (Hou,
Shi, Zhai, & Le, 2007). At the same time, today’s society appears
to be experiencing a trend of ‘green’ consumerism. It means that
food manufacturers should seek new ‘green’ or natural methods to
make food safe. There is recent interest in the development of novel
combinations of natural antimicrobial plant extracts to improve the
quality and safety of agroindustrial products (Periago & Moezelaar,
2001; Periago, Palop, & Fernández, 2001).
Spices and their extracts are generally recognized as safe
(GRAS), because of their traditional use without any documented
detrimental impact (Newton, Lau, & Wright, 2000; Shan, Cai,
Brooks, & Corke, 2007). Several of these spices and their essential
oil extracts have been reported to posses antimicrobial activities.
Cinnamon is traditionally harvested in Asian countries. And it has
biological activities, such as antibacterial, antifungal, insecticidal
and antioxidant properties. The principal constituents of cinnamon
oil, namely cinnamaldehyde, exhibits an antimicrobial effect
against a wide range of microorganisms (Sanla-Ead et al., 2012). In
order to select effective and safe nature antibacterial ingredients, in
this study, more than 30 kinds of plant extracts were selected as
study objects. A standard broth microdilution method was used to
detect their antimicrobial activity, cytotoxicity test was used to
detect their safety and synergy assay were used to detect out which
have synergistic effect. Then, time-kill curves were used to verify
their bactericidal capacity deeply. This study will lay the foundation
for the development of new type of the composite natural food
preservatives.
2. Materials and methods
2.1. Reagents and strains

* Corresponding author. Tel.: þ86 13304325228; fax: þ86 431 87835760. Thirty one kinds of plant extracts were selected as study objects,
E-mail address: linsongyi730@163.com (S. Lin). as presented in Table 1. All plant extracts were dissolved in

0956-7135/$ e see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodcont.2013.05.032
620 H. Ye et al. / Food Control 34 (2013) 619e623

Table 1
The Place of production, Purity and MIC values of 31 kinds of plant extracts.

Name Place of production Purity (%) MIC (mg/mL)

E. coli/S. aureus

Cinnamaldehyde Aladdin Chemistry Co., Ltd. (Shanghai, China) 98.0 0.31/0.31

Curcumin Chinese Institute of Food and Drug Test. (Beijing, China) 98.0 >2.50/>2.50
Citral Baoji City, Best Plant Materials Co., Ltd. (Baoji, China) 97.0 2.50/2.50
P-Anisaldehyde Chinese Institute of Food and Drug Test. (Beijing, China) 98.0 2.50/2.50
Allicin Chinese Institute of Food and Drug Test. (Beijing, China) 88.4 >2.50/>2.50
Andrographolide Chinese Institute of Food and Drug Test. (Beijing, China) 99.0 >2.50/>2.50
Eugenol Chinese Institute of Food and Drug Test. (Beijing, China) 98.0 1.25/1.25
Carvacrol SigmaeAldrich. Inc. (St. Louis, USA) 98.0 0.31/0.31
Cinnamic acid Chinese Institute of Food and Drug Test. (Beijing, China) 98.0 2.50/> 2.50
Citric acid Chinese Institute of Food and Drug Test. (Beijing, China) 99.0 2.50/>2.50
Oleanolic acid JiangXi Hengcheng Natural Spices Oil Co., Ltd. (Ji’an, China) 92.0 0.31/0.63
Perilla oil Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 98.0 2.50/2.50
Thymol Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 96.8 2.50/1.25
Clove oil Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 85.0 2.50/1.25
Peppermint oils Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 70.0 >2.50/>2.50
Naringin Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 95.0 0.31/0.63
Quercetin Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 98.0 0.31/0.63
Linalool Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 82.0 >2.50/>2.50
Geraniol Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 97.0 >2.50/>2.50
Camphor Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 97.0 0.63/0.31
Oregano oil Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 90.0 0.31/0.63
Dehydrocostus lactone Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 98.0 >2.50/>2.50
Usnic acid Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 98.0 2.50/>2.50
Chelerythrine Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 98.0 >2.50/>2.50
Resveratrol Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 98.0 2.50/>1.25
Zingerone Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 95.0 >2.50/>2.50
a-Terpineol Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 65.0e75.0 >2.50/>2.50
Sage oil Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 99.0 >2.50/>2.50
Artemisinin Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 99.0 >2.50/>2.50
Eugenyl acetate Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 98.0 1.25/0.63
Apigenin Jiangxi Kangsheng Tang Pharmaceutical Co., Ltd. (Ji’an, China) 98.0 >2.50/2.50

dimethyl sulfoxide (DMSO) at a concentration of 5 g/L under sterile Beef-extract peptone medium blank was used as a control. The MIC
conditions and stored at 4  C when use. The microbial strains were was defined as the lowest antibiotic or antimicrobial concentration
obtained from the Institute of Zoonosis in Jilin University (Chang- which prevented visible growth (Fazeli et al., 2007). After 24 h in-
chun, P.R. China). All the selected strains include the Gram-positive cubation at 37  C, 10 ml of 3-(4, 5-dimethylthiazol-2-yl)-2, 5-
bacteria and the Gram-negative bacteria, as shown in Table 2. diphenyltetrazolium bromide (MTT) which dissolved in phos-
phate buffer solution (PBS) at a concentration of 5 mg/mL was
2.2. MIC determination added to each well, and then incubated for 4 h at 37  C. MTT, which
is a yellow tetrazole, is taken up into cells by endocytosis or
The resistance of bacteria to the antibiotics and the minimum protein-facilitated mechanism and reduced, mainly by mitochon-
inhibitory concentration (MIC) of natural antimicrobials were drial enzymes, to yield a purple formazan product which is largely
determined by standard broth microdilution (Wiegand, Hilpert, & impermeable to cell membranes, thus resulting in its accumulation
Hancock, 2008). Beef-extract peptone medium was used for the within living cells. While the dead cells don’t have this feature. The
food-borne bacteria. Active cultures were generated by inoculating ability of cells to reduce MTT provides an indication of the mito-
the thawed microbial stock suspensions into nutrient broth fol- chondrial integrity and activity which, in turn, may be interpreted
lowed by overnight incubation at 37  C with shaking. The in- as a measure of viability. Which can be detected by the color using
oculums were adjusted to final concentrations of 1  105 cfu/mL by the naked eye. The MIC was the lowest agent concentration at
comparison with a McFarland No. 1 turbidity standard. The natural which no purple color (loss of metabolic activity) appeared. All tests
antimicrobials were dissolved in beef-extract peptone medium were performed in triplicate.
with DMSO in two-fold serial dilutions from 5 mg/mL to 0.078 mg/
mL. Then, 100 ml of the individual antimicrobials and 100 ml of the 2.3. Synergy testing
1  105 cfu/mL bacteria were dispensed into the 96 well plates.
The checkerboard method used for measurement of interactive
inhibition (Pillai, Moellering, & Eliopoulos, 2005) was determined
Table 2 for synergy between the cinnamaldehyde and carvacrol. In this
MIC values of Cin and Car against 11 food-borne bacteria.
study, the antimicrobial agents were dissolved in beef-extract
Bacteria MIC (mg/mL) Bacteria MIC (mg/mL) peptone medium with DMSO to obtain final concentrations that
Cin Car Cin Car ranged from five dilutions below the 1/2 MIC value obtained from
the MIC determination using two-fold dilutions. The effects of
E. coli 0.31 0.31 S. sanguinis 0.63 0.16
S. aureus 0.31 0.31 E. cloacae 0.63 0.16 combinations were evaluated by calculating the fractional inhibi-
Y. regensburgei 0.31 0.31 S. haemolyticus 0.16 0.31 tory concentration (FIC) index which was combined with the
S. intermedius 0.16 0.31 A. hydrophila 0.31 0.16 checkerboard method (Pei, Zhou, Ji, & Xu, 2009) for each combi-
K. kristinae 0.31 0.31 S. enteritidis 0.31 0.31 nation using the following formula. The FIC was used to interpret
L. garvieae 0.63 0.16
the test results as follows: FIC  0.5, synergy; FIC ¼ 0.5e4, no
 H. Ye et al. / Food Control 34 (2013) 619e623
interaction; FIC > 4.0, antagonism (De Logu et al., 2002). All tests
were performed in triplicate.
FIC ¼ ðMICa combination=MICa aloneÞ
þ ðMICb combination=MICb aloneÞ
2.4. Time-kill assay
The time-kill assay was conducted according to modification of
a previous method (Kent, Bakhtiar, & Shanson, 1992). The number
of viable bacteria was monitored by measuring the optical density
(OD) values at 600 nm on a microplate reader. The time-kill curves
were obtained via plotting the OD values as a function of the in-
cubation time (Bai et al., 2011). Escherichia coli, as indicators of food
microbial contamination, belongs to the gram-negative bacteria;
Staphylococcus aureus belongs to the gram positive coccus, it is a
very common food-born bacteria. So, E. coli and S. aureus were
selected for time-kill curves assay. The MICs for the strain was
determined in beef-extract peptone medium with an inoculum of
approximately 1  105 cfu/mL. Three test series were designed: the
serie of cinnamaldehyde alone, the serie of carvacrol alone, and the
combination serie of the two agents. For the cinnamaldehyde or
carvacrol agent alone, wells of medium were prepared containing
each antimicrobial alone at a concentration equivalent to the MIC;
And for the combination group, wells of medium were prepared at
concentrations equivalent to the 1/2 MIC of each one. MTT was
added in different stated wells at 0, 1, 2, 4, 8 and 12 h of incubation
at 37  C (Erdogan-Yildirim, Burian, Manafi, & Zeitlinger, 2011). OD
values were read at 3 h intervals after addition of MTT. The test
series were all performed in triplicate.
2.5. Cytotoxic assay
The cytotoxicity of cinnamaldehyde and carvacrol were evalu-
ated against Vero cell lines obtained from the American Type Cul-
ture Collection. Cells were cultured at 37  C in a 5% CO2
environment in Dulbecco’s modified Eagle’s medium (Sigma Di-
agnostics), supplemented with 10% fetal calf serum, 2 mM gluta-
mine and 1% penicillin/streptomycin solution. Cells were
resuspended in growth medium at a final concentration of
6  104 cells per well, placed in a 96-well culture microplate
(200 mL per well) and incubated with various concentrations of
cinnamaldehyde and carvacrol for 48 h. Cytotoxicity was deter-
mined using a MTT assay. The IC10 value was calculated, repre-
senting the concentration of the compound that inhibited 10% of
cell growth.
2.6. Statistical analysis
All test series were performed in triplicate. For MIC determi-
nation, synergy testing and cytotoxic assay, the results were
expressed as mean value; For time-kill assay, all OD values were
expressed as mean  SD. Statistical analysis were performed with
the software SPSS 11.5.
3. Results and discussion
3.1. MIC determination
The MIC values of 31 kinds of plant extracts against E. coli and
S. aureus are presented in Table 1. It can be seen that among all
the active components, cinnamaldehyde and carvacrol expressed
the highest antibacterial activities both to E. coli (0.31 mg/mL)
and S. aureus (0.31 mg/mL). So cinnamaldehyde and carvacrol
621
were selected as suitable antibacterial agents for further
investigation.
The MICs of cinnamaldehyde and carvacrol against the 11 bac-
terial strains are presented in Table 2. It was found that for all the
tested bacterial strains, both cinnamaldehyde and carvacrol showed
strong antimicrobial activity. Cinnamaldehyde showed the best
antimicrobial activity against Staphylococcus intermedius and
Staphylococcus haemolyticus, whose MIC values are 0.16 mg/mL.
While carvacrol showed highest antimicrobial activity against
Streptococcus sanguinis, Lactococcus garvieae, Enterobacter cloacae
and Aeromonas hydrophila at 0.16 mg/mL. Cinnamaldehyde and
carvacrol have the same antimicrobial activity toward E. coli,
S. aureus, Yokenella regensburgei, Kocuria kristinae, and Salmonella
enteritidis which all had MICs of 0.31 mg/mL. Cinnamaldehyde and
carvacrol showed high inhibitory activities against most of the
bacteria tested, especially to S. haemolyticus (Cin/Car: 0.16 mg/mL/
0.31 mg/mL), S. intermedius (Cin/Car: 0.16 mg/mL/0.31 mg/mL) and
A. hydrophila (Cin/Car: 0.31 mg/mL/0.16 mg/mL). To some extent,
these results were similar to those of previous works: Palaniappan
and colleague’s study showed that cinnamaldehyde and carvacrol
effectively inhibited Salmonella typhimurium, E. coli, S. aureus and
Streptococcus pyogenes (Palaniappan & Holley, 2010); Sanla-Ead and
colleagues reported that cinnamaldehyde was able to inhibit a wide
spectrum of food pathogens and spoilage microorganisms (Sanla-
Ead et al., 2012); It was also identified that cinnamaldehyde and
carvacrol have the antibacterial activity against Bacillus cereus
(Palaniappan & Holley, 2010; Valero & Francés, 2006). Previous re-
ports indicated that Gram negative bacteria are more resistant to
essential oils (cinnamaldehyde, clove, allicin, onion oil, and oregano
oil) than Gram positive bacteria (Ceylan, & Fung, 2004; Sanla-Ead
et al., 2012; Shan et al., 2007). Results presented here are partially
in agreement with that observation: When E. coli (G),
Y. regensburgei (G), E. cloacae (G), A. hydrophila (G) and
S. enteritidis (G) were exposed to cinnamaldehyde, their MIC values
are all higher than 0.31 mg/mL; while, when they were exposed to
carvacrol, they expressed different resistance (E. coli (G): 0.31 mg/
mL; Y. regensburgei (G): 0.31 mg/mL; E. cloacae (G): 0.16 mg/mL;
A. hydrophila (G): 0.16 mg/mL; S. enteritidis (G): 0.31 mg/mL). In
addition, L. garvieae, S. sanguinis and E. cloacae are more sensitive to
carvacrol (MIC: 0.16 mg/mL) than to cinnamaldehyde (MIC:
0.63 mg/mL), which is different from other reports that state these
bacteria were more sensitive to cinnamaldehyde than to carvacrol
(Sanla-Ead et al., 2012). These differences may be due to the strains
used here were resistant to cinnamaldehyde. Moreover, many fac-
tors could affect the MIC value, such as temperature, inoculum size
and test methods. Besides, factors such as rate of solubility of natural
antimicrobials are difficult to monitor and may result in erroneous
findings (Sanla-Ead et al., 2012).
3.2. Synergy testing
The presence of synergistic interaction was examined by
calculation of FIC values. It can be seen from Table 3 that cinna-
maldehyde not only decreased the MIC of carvacrol (from 0.16e
0.31 mg/mL to 0.078e0.010 mg/mL), but also the MIC of cinna-
maldehyde itself was reduced (from 0.16e0.63 mg/mL to 0.078e
0.009 mg/mL) by carvacrol for the most tested bacteria. The results
showed that cinnamaldehyde and carvacrol had a synergistic
inhibitory effect on 7 kinds of tested bacteria: E. coli (FIC: 0.281),
S. aureus (FIC: 0.281), Y. regensburgei (FIC: 0.281), S. intermedius
(FIC: 0.313), K. kristinae (FIC: 0.156), S. haemolyticus (FIC: 0.313) and
S. enteritidis (FIC: 0.281). Additionally, it is worth noting that all the
FCI values were smaller than 4.0, which indicate that there is no
antagonism between cinnamaldehyde and carvacrol. Based on
these results, it can be accounted that cinnamaldehyde and
622 H. Ye et al. / Food Control 34 (2013) 619e623

Table 3
Combination of cinnamaldehyde and carvacrol.

Bacteria Combination MIC Cin/ Combination FICs Conclusion

Car (mg/mL) (mg/mL)

E. coil 0.010/0.078 0.281 Synergy

S. aureus 0.078/0.010 0.281 Synergy
Y. regensburge 0.010/0.078 0.281 Synergy
S. intermedius 0.010/0.078 0.313 Synergy
K. kristinae 0.010/0.039 0.156 Synergy
L. garvieae 0.156/0.078 0.516 No Synergy
S. sanguinis 0.156/0.078 0.750 No Synergy
E. cloacae 0.313/0.009 0.563 No Synergy
S. haemolyticus 0.010/0.078 0.313 Synergy
A. hydrophila 0.009/0.156 1 < 1.031 < 4 No Synergy
S. enteritidis 0.010/0.078 0.281 Synergy

carvacrol have synergistic antimicrobial activities at least for most

of the test bacteria.

3.3. Time-kill assay

To further prove the synergistic antimicrobial effect of the two

reagents, a time-kill curve study was conducted using E. coli and
S. aureus. The smaller the OD value, the higher the death rate. After
12 h of incubation, for both strains, the combination sample, which
were exposed to cinnamaldehyde and carvacrol simultaneously,
not only showed smaller OD values compared with the cinna-
maldehyde and carvacrol alone, but also displayed a rapid declining
trend. That is to say, the combination sample showed better
bactericidal effect compared with the cinnamaldehyde and carva-
crol alone, as shown in Fig. 1. The exact synergistic mechanism of
the natural antimicrobials is presently unknown; however, it is
likely due to some structural changes in the bacteria. For example,
the natural antimicrobials may have facilitated penetration of the
drug through the outer layers of the bacterial cell wall or acted by
blocking the inhibitory effects of protective enzymes, or interfered
with single or multiple metabolic targets of the antibiotic
(Palaniappan & Holley, 2010). Wendakoon and Sakaguchi thought Fig. 1. Time-kill curves for E. coil and S. aureus in the presence of individual and
that the cinnamaldehyde may inhibit amino acid decarboxylase combination of antimicrobials: (a) E. coil; (b) S. aureus. - 0.31 mg/mL carvacrol; C
0.31 mg/mL cinnamaldehyde; : 0.16 mg/mL carvacrol and 0.16 mg/mL cinnamalde-
activity to bind proteins (Wendakoon, & Sakaguchi, 1995); Similar
hyde. Data are mean  S.D. values.
results were reported by Pol, Mastwijk, Slump, Popa, and Smid
(2001) where they also believed that the antibacterial mechanism
of cinnamaldehyde wasn’t disintegrating the outer membrane or 2009), and the outer layers of Gram-negative and Gram-positive
depleting the intracellular ATP pool. While, the antibacterial bacteria are significantly different (Kalemba, & Kunicka, 2003).
mechanism of carvacrol is generally considered to be reducing the That is a possible explanation for these observations about the
membrane concentrations of lipophilic compounds: Several studies difference sensitivity between Gram negative and Gram positive
considered that the carvacrol (lipophilic hydrocarbons) accumulate bacteria (Shan et al., 2007).
in the membrane lipid bilayer, affecting the structural and func-
tional properties of these membranes. As a result of accumulated 3.4. Cytotoxic assay
hydrocarbon molecules, the membrane loses its integrity, and an
increase in permeability to protons and ions may happen. Conse- In terms of cytotoxicity, cinnamaldehyde showed an IC10 value
quently, dissipation of the proton motive force (PMF) and impair- of 6.3 mg/mL and carvacrol showed an IC10 value of 12.6 mg/mL in
ment of intracellular pH homeostasis may occur. At the same time, Vero cells. The results of the cytotoxicity study of cinnamaldehyde
the effects on the membrane-embedded proteins probably result to and carvacrol suggest that they appear to not have significant or
a large extent from changes in the lipid environment (Cosentino marginal effects in vitro while they exhibited mild to moderate
et al., 1999; Davidson, 1997; Panizzi, Flamini, Cioni, & Morelli, toxic effects at the cellular level. Furthermore, the toxicity data
1993; Sikkema, De Bont, & Poolman, 1995; Sivropoulou, Kokkini, appears not to raise concern in view of the present levels of use,
Lanaras, & Arsenakis, 1995). Overall, the antibacterial mechanism which was consistent with the common sense spices that have
of carvacrol is probably to be the disturbance of the cytoplasmic been used safely since ancient times as food flavoring agents and
membrane, disrupting the PMF, electron flow, active transport and are now mainly considered “generally regarded as safe” (GRAS)
coagulation of cell contents (Di Pasqua, Hoskins, Betts, & Mauriello, (Fazeli et al., 2007).
2006). Maybe the difference in the antibacterial mechanism of
cinnamaldehyde and carvacrol is just the reason for the cinna- 4. Conclusions
maldehyde/carvacrol combination’s synergism. Additionally, the
mechanism of antibacterial action might be different and the In this study, it can be concluded that: Cinnamaldehyde and
combined action might vary with different bacteria (Pei et al., carvacrol not only exhibit high antibacterial activities and have
 H. Ye et al. / Food Control 34 (2013) 619e623
synergistic antimicrobial activities at least for most of the test
bacteria, but also are safe as food preservatives. The results indi-
cated that cinnamaldehyde and carvacrol may serve as promising
naturally sourced composite food preservatives. Additionally,
further studies are needed to investigate the mechanism and
relationship between the antibacterial activity and chemical
structure of cinnamaldehyde and carvacrol.
Acknowledgments
The authors acknowledge the financial support provided by the
Projects in the National Natural Science Foundation (31101345). We
gratefully thank the Institute of Zoonosis in Jilin University for
supply of the food-borne microbial strains.
Appendix A
Generally recognized as safe (GRAS)
Minimum inhibitory concentration (MIC)
Dimethyl sulfoxide (DMSO)
3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide
(MTT)
Phosphate buffer solution (PBS)
Fractional inhibitory concentration (FIC)
Optical density (OD)
Standard deviation (SD)
Proton motive force (PMF)
Cinnamaldehyde (Cin)
Carvacrol (Car)
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