International Journal of Food Microbiology 111 (2006) 6 – 11

www.elsevier.com/locate/ijfoodmicro

Antimicrobial effect of spices and herbs on Vibrio parahaemolyticus

Yutaka Yano ⁎, Masataka Satomi, Hiroshi Oikawa
Seafood Safety Section, National Research Institute of Fisheries Science, 2-12-4 Fukuura, Yokohama 2368648, Japan
Received 2 May 2005; received in revised form 1 March 2006; accepted 25 April 2006

Abstract

The antimicrobial effects of spices and herbs from 18 plant species were examined on a foodborne pathogen, Vibrio parahaemolyticus, with
the use of combinations of temperatures and nutrient levels. Basil, clove, garlic, horseradish, marjoram, oregano, rosemary, and thyme exhibited
antibacterial activities at incubation of 30 °C, while with the exception of horseradish, the same spices and additional 7 species exhibited the
activities at 5 °C. The lowest MIC (minimum inhibitory concentration) was 0.125% observed in clove and marjoram at 30 °C in a nutrient rich
medium. Lowering of incubation temperature produced little effect on the MICs except for turmeric. The decreasing of the MIC in turmeric
appeared to be basically attributed to the sensitivity of the bacterium to coldness. In nutrient poor medium, the lowest was 0.001 and 0.00025% in
marjoram at 30 °C and at 5 °C, respectively. The sensitivity to several spices and herbs was similar among different clinical serotypes including the
emerging strain O3:K6. These results suggest that the spices and herbs can be practical for protecting seafood from the risk of contamination by V.
parahaemolyticus and used in hurdle technology with low temperature.
© 2006 Elsevier B.V. All rights reserved.

Keywords: Vibrio parahaemolyticus; Spices; Herbs; Antibacterial activity

1. Introduction pathogenic bacteria (Deans and Ritchie, 1987; Shelef et al.,

Vibrio parahaemolyticus is a bacterium which causes mild
gastroenteritis in humans on consumption of infected seafood
(Oliver and Kaper, 1997). The bacterium generally inhabits
coastal environments in tropical and temperate zones, and con-
taminates fishery products caught in these areas, especially
during warm periods. In Japan, a variety of seafood has been
traditionally consumed, and raw or lightly cooked seafood is
favored. This eating habit seems to provide an explanation for
many cases of foodborne disease by V. parahaemolyticus in the
country. The foodborne diseases by the bacterium have recently
also occurred in other regions of the world with the emerging of
new pandemic clone O3:K6 and often has occurred on eating
seafood (Matsumoto et al., 2000; Wong et al., 2000; Depaola
et al., 2000). Some efforts are needed to enhance the safety of
seafood.
Antimicrobial activities of spices and herbs and essential oils
have been well known for long time. Many studies reported the
activities of spices and herbs or essential oils to foodborne
⁎ Corresponding author. Tel.: +81 45 788 7669; fax: +81 45 788 5001.
E-mail address: yanoya@affrc.go.jp (Y. Yano).
0168-1605/$ - see front matter © 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.ijfoodmicro.2006.04.031
1980; Hirasa and Takemasa, 1998). A Japanese spice, wasabi
(Wasabi japonica), is traditionally used on eating raw fish such
as sushi in Japan. The spice has been known to have antimic-
robial effect against several bacteria including V. parahaemo-
lyticus and is believed to contribute to the safety of eating raw
seafood (Hasegawa et al., 1999). However, a few studies have
investigated the effects of other spices and herbs against marine
pathogenic bacteria such as V. parahaemolyticus (Beuchat, 1976;
Koga et al., 1999).
The Japanese style of cuisine of eating raw and lightly
cooked seafood is increasingly popular in Europe and the
United States, and in Asian countries, and seems to be also
adopted in the local cuisines with globalization of food.
Many kinds of spices and herbs have been used for taste and
preservation of various food and cuisine in the world and
could be introduced to raw and lightly cooked seafood. For
example, in Japan, raw fish slices with raw vegetables are
covered with dressing (seasoning) containing spices and
herbs like carpaccio in Italian cuisines and such dressings are
going to be commercial. Thus, in this study, we screened the
potential of whole spices and herbs to inhibit V. parahaemo-
lyticus, investigated the minimum inhibitory concentrations
 Y. Yano et al. / International Journal of Food Microbiology 111 (2006) 6–11
(MICs) of these spices and herbs under different nutrient and
thermal conditions, and compared the activities among dif-
ferent serotypes of the bacterium.
2. Materials and methods
2.1. Spices and herbs
Dry spices and herbs for food were purchased from gro-
ceries and processing companies: anise (Pimpinella anisum),
basil (Ocimum basilicum), carom (Trachyspermum ammi),
clove (Syzygium aromaticum), coriander (Coriandrum sati-
vum), cumin (Cuminum cyminum), garlic (Allium sativum),
ginger (Zingiber officinale), horseradish (Armoracia rustica-
na), Japanese pepper (Zanthoxylum piperitum), marjoram
(Origanum marjorana), oregano (Origanum vulgare), pep-
permint (Mentha piperita), rosemary (Rosmarinus officina-
lis), sage (Salvia officinalis), spearmint (Mentha spicata),
thyme (Thymus vulgaris), and turmeric (Curcuma longa).
Several spices and herbs were obtained from different pro-
cessing companies. Each spice and herb was finely ground in
a mortar and sterile distilled water was added to make a
concentration of 5% (weight of dry matter/volume of water).
The suspensions were kept for 24 h at room temperature for
hydration and extraction, and then stored until experiments at
− 20 °C. Prior to the tests, viable bacterial counts in the
suspensions were examined with Marine agar (MA; Difco,
USA) and thiosulfate–citrate–bile–sucrose agar (TCBS; Nis-
sui Pharmacy, Japan) plates at 37 °C incubation.
2.2. Bacterial strains
V. parahaemolyticus clinical strain VPY01 (serotype, O3:K6)
which was positive in thermo stable direct hemolysin (TDH)
production was used in all experiments. Clinical strains VPY12
(O3:K6), VPY21 (O4:K8), and VPY22 (O5:K68) which were
positive in TDH production, and environmental strains A6 and
B1 which were negative in PCR for tdh and trh genes were also
used in this study. For the experiments, a loopful of the working
stocks was transferred to 3 ml of heart infusion broth (HI broth;
Difco) with NaCl added at a final concentration of 3% (Na–HI
broth) and incubated at 37 °C for 18 h. The overnight cultures
were used for the experiments. Escherichia coli non-pathogenic
strain JCM109 was used in this study. E. coli was cultured using
the same protocol for V. parahaemolyticus except for using HI
broth only as the culture medium.
2.3. Screening of spices and herbs
The antibacterial activity of spices and herbs was screened
in nutrient rich medium (Na–HI broth). A 100 μl aliquot of
5% suspension of each spice or herb in Na–HI broth was put
in each well of a 96 well micro-titer plate and kept for 30 min
at 5 or 30 °C. The overnight culture of V. parahaemolyticus
strain VPY01 was diluted with fresh Na–HI broth to a level
of 104 CFU/ml. Immediately, a 100 μl aliquot of the bacterial
dilution was inoculated to each well of the plates, and then
7
incubated at 5 or 30 °C for 24 h. For a non-treated control,
the bacterial dilution was inoculated to the same amount of
Na–HI broth. In addition, fresh Na–HI broth was added to an
equal volume of the 5% suspensions and incubated to check
the background bacterial counts in the spices and herbs. After
incubation, a 100 μl aliquot of the mixed solutions and of the
10-fold dilutions was spread onto TCBS agar plate and in-
cubated at 37 °C for 18 h. The antibacterial activities of the
spices and herbs against E. coli were also examined for com-
parison using HI broth, and the survival after the treatment
was examined on DHL plates. Viable counts in non-treated
control of V. parahaemolyticus were reduced by 1-log with
the incubation at 5 °C for 24 h in this study. Viable counts in
the control of E. coli were constant or increased only 0.1-log
with the same incubation. Thus, the antibacterial activities to
these bacteria at 5 °C were difficult to be evaluated. In this
study, the absence of culturable bacteria after a 24 h incu-
bation was defined as positive for the antibacterial activity.
2.4. Determination of MICs
Each well of a sterile 96 well micro-titer plate was filled with
a 100 μl aliquot of Na–HI broth. The first column of wells in
each micro-titer plate received a 100 μl of the broth containing
the spices or herbs to be tested. After mixing by pipetting, the
mixtures (100 μl) were transferred to the next column of wells
in a process of 1:1 serial dilution until column #12. Then, the
plates were incubated at 5 or 30 °C for 30 min before bacterial
inoculation. An inoculum was prepared by diluting the over-
night culture of V. parahaemolyticus strain VPY01 to a level of
2 × 104 CFU/ml. Each well in the micro-titer plate was inocu-
lated with a 100 μl of the inoculum and incubated at 5 or 30 °C
for 24 h. Growth or survival of the bacterium after incubation
was examined with following method because the bacterium
does not grow at 5 °C. A 100 μl of the mixture from each well
was spread onto TCBS and incubated at 37 °C for 24 h. The
concentration in the lowest serial dilution of the spices and
herbs at which growth did not occur on TCBS was recorded as
the minimal inhibitory concentration (MIC). The MICs were
also determined in sterile natural seawater (NSW) instead of
Na–HI broth.
2.5. Changes of viable counts in the media added marjoram
and turmeric
The change of viable counts of V. parahaemolyticus strain
VPY01 with the addition of marjoram was investigated. Over-
night culture of the bacterium in Na–HI broth was diluted with
Na–HI at a level of 105 CFU/ml. The bacterial solutions were
kept at 5 or 30 °C for 30 min to adjust the temperatures,
marjoram was added at concentrations of 0.063 to 1%, and
incubated at 5 and 30 °C. The bacterial samples were collected
at 3, 6, 9, and 24 h after the initiation of incubation, and diluted
10-fold in PBS. The dilutions were spread on TCBS and MA,
incubated at 37 °C overnight, and then the colonies were
counted. The survival curve was also examined in the medium
with turmeric added using the same manner.
8 Y. Yano et al. / International Journal of Food Microbiology 111 (2006) 6–11

2.6. Comparison of sensitivity among V. parahaemolyticus
strains
The sensitivity to several spices and herbs was compared
among the clinical and environmental strains of V. parahaemo-
lyticus. The MICs were determined at 30 °C incubation for
clove, marjoram, oregano and rosemary, and at 5 °C incubation
for turmeric in Na–HI broth using the same procedure as
described above. The sensitivity to each spice and herb was
defined as the reciprocal of the MIC.
3. Results
3.1. Screening of antibacterial activity of spices and herbs
Twenty three spices and herbs of 18 species were examined
for their antibacterial activities against V. parahaemolyticus at a
final concentration of 2.5% (Table 1). In the incubation at 30 °C,
basil, clove, garlic, horseradish, marjoram, oregano, rosemary,
and thyme showed antibacterial activities. By decreasing the
incubation temperature to 5 °C, carom, ginger, Japanese pepper,
peppermint, sage, spearmint, and turmeric were additionally
found to have antibacterial activities, but horseradish lost the
activity. In these tests, the viable counts of the bacterium were
constant or reduced by 1-log after the incubation of low
temperature (data not shown). Antibacterial activities against E.
coli were detected for basil, clove, garlic, marjoram, oregano,
and rosemary at 30 °C and for all of the same but basil and garlic
at 5 °C.
3.2. Minimum inhibitory concentrations of spices and herbs
MICs of basil, clove, garlic, horseradish, marjoram, oregano,
rosemary, thyme, and turmeric were determined under several
conditions (Table 2). In the nutrient rich medium (Na–HI
broth), the lowest MIC at 30 °C was 0.125% in clove and
marjoram, and at 5 °C was 0.063% in marjoram and turmeric. In
other spices and herbs, the MICs ranged from 0.5% to >2.0% at
30 °C and 0.25% to > 2.0% at 5 °C. Reducing the incubation
temperature produced little effect on the MICs of the spice and
herbs except for turmeric, but the MIC of turmeric decreased
from > 2% at 30 °C to 0.063% at 5 °C. In the nutrient poor
medium (NSW), the MICs of spices and herbs were generally
lower than those in the nutrient rich medium when compared
between the same temperatures. Especially, the MICs of basil
and marjoram were lowered to 0.016 and 0.001% in the poor
medium at 30 °C incubation, respectively. At 5 °C incubation,
the MIC of marjoram was much lowered to 0.00024% in the

Table 1
The results of screening for antibacterial activities of spices and herbs against Vibrio parahaemolyticus and Escherichia coli
Spices and Scientific name pHa Growth or survival after 24 h incubation
herbs
V. parahaemolyticus E. coli
30 °Cb 5 °C 30 °C 5 °C
c
Anise seed Pimpinella anisum 5.7 + ± + ±
Basil-I Ocimum basilicum 5.9 − − − ±
Basil-II Ocimum basilicum 6.0 − − − ±
Carom seed Trachyspermum ammi 5.7 + − + ±
Clove Syzygium aromaticum 3.6 − − − −
Coriander seed Coriandrum sativum 4.7 + ± + ±
Cumin seed-I Cuminum cyminum 5.8 + ± + ±
Cumin seed-II Cuminum cyminum 5.7 + ± + ±
Garlic Allium sativum 5.8 − − − ±
Ginger Zingiber officinale 5.7 + − + ±
Horseradish Armoracia rusticana 4.3 − ± + ±
Japanese pepper Zanthoxylum piperitum 4.8 + − + ±
Marjoram Origanum marjorana 5.9 − − − −
Oregano-I Origanum vulgare 5.6 − − − −
Oregano-II Origanum vulgare 5.4 − − − −
Peppermint Mentha piperita 6.2 + − + ±
Rosemary Rosmarinus officinalis 5.8 − − − −
Sage Salvia officinalis 6.0 + − + ±
Spearmint Mentha spicata 5.7 + − + ±
Thyme-I Thymus vulgaris 5.8 − − + ±
Thyme-II Thymus vulgaris 5.8 − − + ±
Turmeric-I Curcuma longa 6.1 + − + ±
Turmeric-II Curcuma longa 5.8 + − + ±
Control 7.2 + ±d + ±e
a
The pHs were determined in 10% solutions of the spices and herbs.
b
Incubation temperature in the screening tests.
c
+; Growth, viable counts increased more than 106 CFU/ml after 24 h incubation. ±; Survival, viable counts were from 102 to 105 CFU/ml after 24 h incubation.
−; Death, viable counts were reduced less than 101 CFU/ml after 24 h incubation.
d
Viable counts were reduced approximately by 1-log after 24 h incubation.
e
Viable counts were constant after 24 h incubation.
 Y. Yano et al. / International Journal of Food Microbiology 111 (2006) 6–11 9

Table 2
Minimum inhibitory concentrations (%) of spices and herbs against Vibrio
parahaemolyticus
Spices and Na-HI brotha NSWb
herbs
30 °Cc 5 °Cd 30 °C 5 °C
Basil 2 1 0.016 1
Clove 0.125 0.25 0.004 0.016
Garlic 1 1 0.25 2
Horseradish 2 >2 1 1
Marjoram 0.125 0.063 0.001 0.00025
Oregano 0.5 0.5 0.032 0.25
Rosemary 0.5 0.25 0.008 0.063

▪
Thyme 2 1 0.032 0.5 Fig. 2. Changes of viable counts of V. parahaemolyticus in Na–HI broth with
Turmeric >2 0.063 2 0.004 turmeric added at concentrations of ○: 1%, ▵: 0.25%, □: 0.063%, and : 0%
a
Heart Infusion broth added NaCl. (control).
b
Sterile natural sea water.
c
Incubation temperature.
d concentration of 1%, the bacterial level decreased to below
At 5 °C incubation, the viable counts in the broth with no spices and herbs
(control) were constant or reduced by 1-log after 24 hr.
detection limit within 10 min. Although the decrease of the
population at 5 °C was slower than at 30 °C, the population was
reduced to below the detection limit in cultures added with
nutrient poor medium. When the MICs in the nutrient poor marjoram at all concentrations including 0.063% within 24 h. In
medium were compared between both temperatures, most of the the broth added turmeric (Fig. 2), the growth was delayed at
spices and herbs exhibited higher MICs at 5 °C than at 30 °C. concentrations of 1% and 0.25% when incubated at 30 °C but
For example, the MIC of basil at 5 °C was 64 times higher than not stopped. On the other hand, the bacterial counts decreased
at 30 °C. On the other hand, the MIC of turmeric at 5 °C was one just after addition at concentrations of 1 and 0.25%, and 3 h at a
five-hundreds-twelfth of that at 30 °C. concentration of 0.063% when incubated at 5 °C. The bacterial
counts were reduced less than detection limit at all concentra-
3.3. Changes of viable counts in media with marjoram added tions for 24 h incubation.

The inhibitory effect of marjoram and turmeric against V.
parahaemolyticus was confirmed in Na–HI broth (Fig. 1). In
the Na–HI broth without marjoram (control), the bacterium
which was inoculated at a level of 105 CFU/ml started to grow
exponentially at 3 h and reached a level of 108 CFUs/ml at 9 h at
30 °C. In the broth with marjoram added and incubated at 30 °C,
the bacterial level decreased below the detection limit within
10 min at concentrations of 1% and 0.25% and within 3 h at that
of 0.125%. The addition at a concentration of 0.063% caused
the delay of growth but did not stop the growth finally. At 5 °C
incubation, the viable counts gradually decreased even in the
control broth, and 1-log and 1.5-log decreases were observed at
9 and 24 h, respectively. With the addition of marjoram at a
3.4. Comparison of sensitivity among V. parahaemolyticus
strains
The sensitivity to spices and herbs was compared among
clinical strains with different serotypes and environmental strains
(Fig. 3). The sensitivity to each spice and herb was defined as the
reciprocal of the MIC in the present study. It was generally
similar among the strains, except for turmeric, and the difference
was less than 2 times among the strains. In these tests, the
inoculum sizes did not exceed 1-log among the strains used (data
not shown). In addition, increasing by one order of magnitude in
inoculum size did not affect the MICs or increased by only 2

Fig. 3. Variation in sensitivity of different strains of Vibrio parahaemolyticus to

spices and herbs. The sensitivity is expressed as the reciprocal of the minimum
Fig. 1. Changes of viable counts of V. parahaemolyticus in Na–HI broth with inhibitory concentration (%). Strain VPY01 (O3:K6, tdh+), strain VPY12 (O3:

▪
marjoram added at concentrations of ○: 1%, ▵: 0.25%, ▴: 0.125%, □: 0.063%, K6, tdh+), strain VPY21 (O4:K8, tdh+), strain VPY22 (O5:K68, tdh+), strain
and : 0% (control). A6 (environmental, tdh−), and strain B1 (environmental, tdh−).
10 Y. Yano et al. / International Journal of Food Microbiology 111 (2006) 6–11
Table 3
Effect of inoculum size on minimum inhibitory concentrations (%) against
Vibrio parahaemolyticus
Inoculuma Na-HI brothb NSWc
Clove Marjoram Clove Marjoram
1.0 × 105 0.125 0.061 0.008 0.0005
1.0 × 104 0.061 0.061 0.008 0.0005
1.0 × 103 0.061 0.031 0.004 0.00025
a
The bacterial number treated by the spices.
b
Heart Infusion broth added NaCl.
c
Sterile natural seawater.
times the MICs (Table 3). The sensitivity to turmeric was rela-
tively variable in the strains and the largest difference was more
than 10 times.
4. Discussion
Basil, clove, garlic, horseradish, marjoram, oregano, rose-
mary, and thyme were found to exhibit inhibitory activities
against V. parahaemolyticus in the incubation with nutrient rich
medium and 30 °C which was good for the growth of the
bacterium. Previous studies have reported the presence of anti-
bacterial activities of oregano, rosemary, and thyme (Beuchat,
1976; Shelef et al., 1980). While antibacterial activity against V.
parahaemolyticus has been unknown in basil, clove, and mar-
joram, these spices and herbs, and essential oils have been well
known to have inhibitory effects against a variety of bacteria
including Gram-negative bacteria (Deans and Ritchie, 1987;
Suppakul et al., 2003). The results in the present study also
indicate that these spices and herbs inhibited the growth of E.
coli. The screening test was performed also at low temperature,
and carom seed, ginger, Japanese pepper, sage, spearmint, and
turmeric were additionally found to exhibit antibacterial activi-
ties against V. parahaemolyticus. This bacterium is well known
to be sensitive to coldness (Oliver and Kaper, 1997) and, in fact,
the viable counts were reduced by 1-log in the control broth
with 5 °C incubation in the tests. Thus, these results don't
indicate that these spices and herbs have antibacterial activities
which are activated by low temperature. These spices and herbs
seem to decrease the survival of the bacterium at low
temperature. Although the following tests found that these
spices and herbs except for turmeric had relatively high MICs at
low temperature incubation, indicating weak antimicrobial
activity, low temperature screening was effective for detecting
the activity against the bacterium, compared with E. coli.
The lowest MIC was 0.125% observed in clove and marjoram
when tested in the nutrient rich medium and at 30 °C. These spices
and herbs had 4 times stronger activities than oregano and rose-
mary which had the second lowest MICs in the tests. Beuchat
(1976) reported that V. parahaemolyticus was highly sensitive to
oregano at 35 °C in a nutrient rich medium. Shelef et al. (1980)
reported that the growth of the bacterium was inhibited by
rosemary at concentrations of 0.3 to 0.5% at 32 °C incubation.
Essential oils of clove as well as those of oregano and rosemary
are well known to have strong antimicrobial effects to a variety of
bacteria, although marjoram relatively had weak activity against
Gram-negative bacteria (Smith-Palmer et al., 1998; Ueda et al.,
1982). The oil of marjoram was also shown to be inhibitory to
bacteria of the family Vibrio, while the activity was lower than
oregano and clove (Deans and Ritchie, 1987). The present study
suggests that marjoram itself has a strong activity against the
bacterium at the same level as clove.
Seafood, which is the main source in foodborne disease by V.
parahaemolyticus, is generally stored in a refrigerator even for a
short time, because seafood easily deteriorates in quality like its
color and flavor. The major antibacterial components of spices
and herbs have been known to be terpenes such as eugenol and
carvacrol (Davidson, 1997). These are generally less dissolved at
low temperature and the antibacterial activity of spices and herbs
may be reduced when used in refrigerators. Previous studies have
reported that temperature effects varied with the kind of spices
and herbs, and bacteria tested (Ting and Deibel, 1991; Burt and
Reinders, 2003). In the present study, decreasing the incubation
temperature generally produced little effect to the MICs, or re-
duced MICs in the nutrient rich medium. The results seem to be
basically attributed to the sensitivity of the bacterium to low
temperature described above, and it is difficult to evaluate the true
activity of the spices and herbs at low temperature. It is, however,
obvious that these spices and herbs decreased the survival of the
bacterium at low temperature. Figs. 1 and 2 also indicated that the
viable counts were much more reduced in the broth added spices
and herbs than those in the control broth at 5 °C. In other words,
the spices and herbs can enhance the destructive effects of low
temperature to the bacterium. The present study suggests that
some of these spices and herbs can be practical for protecting
seafood from the risk of the bacterium and can be used in hurdle
technology with low temperature. The MIC of turmeric was as
low as that of marjoram at low temperature in the nutrient rich
medium. Little has been examined on the antibacterial activity of
turmeric, although the spice was reported to exhibit weak anti-
microbial activity against several bacteria (Lutomski et al., 1974).
In the present study also, the MIC of turmeric was more than 2%
at 30 °C, and that of marjoram was 0.125% at high temperature.
The results in Figs. 2 and 3 also indicated the difference of both
spices in temperature effect of the antibacterial activity. These
results suggest the presence of anti-Vibrio substances in turmeric
which may be different from the ones in marjoram. The anti-
bacterial activity of turmeric may be able to act synergistically
with those of other spices and herbs. Further study is needed to
understand the nature of the antibacterial activity of spices and
herbs.
Spices and herbs are generally applied to food which is a
nutrient rich environment for the bacterium. The antibacterial
activity, however, could be also used in nutrient poor environ-
ments, for example, cleaning of food processing devices and
depuration of shellfish (Birkenhauer and Oliver, 2003). In this
study, the MICs of spices and herbs were generally decreased in
NSW than those in Na–HI broth. The growth of V. parahaemo-
lyticus delays in NSW at 30 °C, compared with Na–HI broth. In
addition, some substances like fatty acids in broth are known to
protect the bacteria from attack of the antimicrobial activity of
spices and herbs (Keweloh et al., 1991). Thus, the MICs were
expected to be decreased in the nutrient poor medium, but the very
 Y. Yano et al. / International Journal of Food Microbiology 111 (2006) 6–11
low MIC of marjoram was interesting. The spice can decrease the
bacterial survival even at a concentration of 2.5 ppm at low
temperature. The MICs of other spices and herbs were increased
with the decreasing of temperature except for marjoram and
turmeric, and the increase may be due to the lower solubility of
antibacterial substances such as terpenes at low temperature.
Marjoram and turmeric may contain substances differing from
those of others, or could contain any nutrients for the bacterium at
high temperature. These spices and herbs could be used for the
control of V. parahaemolyticus in nutrient poor environments.
The presence of various serotypes is known in the bacterium
V. parahaemolyticus. In Japan, serotype O4:K8 was the main
causative agent of the infections in 1994, and since 1996 the
emerging pathogenic strain O3:K6 has been responsible for
many of the recent outbreaks (Matsumoto et al., 2000). The
strain also caused outbreaks in Asia, India, and North America
(Wong et al., 2000; Depaola et al., 2000). In the present study,
the pathogenic serotype strains including O3:K6 and O4:K8
were sensitive to a similar extent in each spice or herb except for
turmeric. Shelef et al. (1980) reported that the pathogenic strains
O4:K8 and O4:K11 were more sensitive to rosemary than a non-
pathogenic one. Hasegawa et al. (1999) showed that pathogenic
strains including O4:K8 were of similar sensitivity, but did not
examine the emerging strain O3:K6. The present study indicates
that the strain O3:K6 is possibly as sensitive as the strains
reported before. Some spices and herbs could contribute to
reducing the risk of the emerging pathogenic strain O3:K6.
Acknowledgements
This study was carried out with financial support (MAFF)
from the Ministry of Agriculture, Forestry, and Fisheries. We
thank Mrs. Y. Shimono and N. Hatano for technical assistance.
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