Toxicity of Plant Essential Oils to Acaricide-Susceptible and

-Resistant Tetranychus urticae (Acari: Tetranychidae) and

Neoseiulus californicus (Acari: Phytoseiidae)
Author(s): Jun Han , Byeoung-Ryeol Choi , Sang-Gyeu Lee , Soon Il Kim , and
Young-Joon Ahn
Source: Journal of Economic Entomology, 103(4):1293-1298. 2010.
Published By: Entomological Society of America
DOI: http://dx.doi.org/10.1603/EC09222
URL: http://www.bioone.org/doi/full/10.1603/EC09222

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 HORTICULTURAL ENTOMOLOGY

Toxicity of Plant Essential Oils to Acaricide-Susceptible and -Resistant

Tetranychus urticae (Acari: Tetranychidae) and Neoseiulus californicus
(Acari: Phytoseiidae)
JUN HAN, BYEOUNG-RYEOL CHOI,1 SANG-GYEU LEE,1 SOON IL KIM,2 AND YOUNG-JOON AHN3
WCU Biomodulation Major, Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921,
Republic of Korea

J. Econ. Entomol. 103(4): 1293Ð1298 (2010); DOI: 10.1603/EC09222

ABSTRACT The toxicity of 10 plant essential oils to adults of acaricide-susceptible, chlorfenapyr-
resistant (CRT-53), fenpropathrin-resistant (FRT-53), pyridaben-resistant (PRT-53), and abamectin-
resistant (ART-53) strains of Tetranychus urticae Koch (Acari: Tetranychidae) and to female Neo-
seiulus californicus McGregor (Acari: Phytoseiidae) was examined using spray or vapor-phase
mortality bioassays. In bioassay with the susceptible adults, lemon eucalyptus (19.3 ␮g/cm3) was the
most toxic oil, followed by peppermint, citronella Java, thyme red, caraway seed, clove leaf, and
pennyroyal oils (LC50, 20.6 Ð23.7 ␮g/cm3). The toxicity of these oils was almost identical against adults
from either of the susceptible and resistant strains, even though CRT-53, FRT-53, PRT-53, and ART-53
adults exhibited high levels of resistance to chlorfenapyr (resistance ratio [RR], ⬎9,140), fenpropath-
rin (RR, 94), pyridaben (RR, ⬎390), and abamectin (RR, 85), respectively. Against female N.
californicus, lemon eucalyptus (LC50, 21.4 ␮g/cm3) was the most toxic oil, whereas the LC50 values
of the other nine oils ranged from 23.2 to 72.6 ␮g/cm3. N. californicus was 1Ð2 times more tolerant than
T. urticae to the test essential oils. Thus, these essential oils merit further study as potential acaricides
for the control of acaricide-resistant T. urticae populations as fumigants.

KEY WORDS botanical acaricide, natural fumigant, essential oil, Tetranychus urticae, Neoseiulus
californicus

The twospotted spider mite, Tetranychus urticae Koch
(Acari: Tetranychidae), is the most economically im-
portant tetranychid mite in many cropping systems
such as fruits, vegetables, ornamentals, and medicinal
plants in Korea (Choi et al. 1989). Control of T. urticae
populations has been provided principally by the use
of chlorinated hydrocarbon (dicofol), organotin (azo-
cyclotin and fenbutatin oxide), mitochondrial elec-
tron transport inhibitor (METI) (fenpyroximate and
pyridaben), pyrethroid (acrinathrin and fenpropath-
rin), and avermectin acaricides. Although still effec-
tive, their continued or repeated use has disrupted
natural biological control systems and led to resur-
gence of this spider mite (Penman and Chapman 1988,
Lee 1990) and often resulted in the development of
resistance (Cho et al. 1995, Devine et al. 2001, Nauen
et al. 2001). Particularly, increasing levels of resistance
to the commonly used acaricides have resulted in
multiple treatments and excessive doses, fostering se-
rious environmental and human health concerns.
Therefore, T. urticae has been the target of an inte-
1 National Academy of Agricultural Science, Rural Development
Administration, Suwon 441-707, Republic of Korea.
2 NaturoBiotech Research Institute, NaturoBiotech Company, Ltd.,
Suwon 441-853, Republic of Korea.
3 Corresponding author, e-mail: yjahn@snu.ac.kr.
grated pest management (IPM) program in which
natural enemies and selective mite control agents are
key components in Þeld and greenhouses. Decreasing
efÞcacy and increasing concern over adverse effects of
the currently used acaricides substantiate the need for
the development of selective T. urticae control alter-
natives to establish an efÞcient resistance manage-
ment strategy based on all available information on the
extent and nature of resistance.
Plant essential oils have been suggested as alterna-
tive sources for mite control products. This approach
is appealing, in part, because they are a rich source of
bioactive chemicals that often produce only minor
adverse effects and often act at multiple and novel
target sites, thereby reducing the potential for resis-
tance (Singh and Upadhyay 1993, Isman 2000, Ahn et
al. 2006, Isman 2006). In addition, plant essential oils
are widely available with some being relatively inex-
pensive compared with plant extracts (Isman 2006).
These potential new acaricides can be applied to Þeld
and greenhouse crops in the same manner as conven-
tional acaricides. Thus, much effort has been focused
on them as potential sources of commercial acaricidal
products largely because certain plant essential oil
preparations and their constituents meet the criteria
of minimum risk pesticides (USEPA 1996, 2009), In

0022-0493/10/1293Ð1298$04.00/0 䉷 2010 Entomological Society of America

1294 JOURNAL OF ECONOMIC ENTOMOLOGY
particular, we initially reported that some essential
oils, such as caraway seed, citronella Java, lemon eu-
calyptus, pennyroyal, and spearmint, had adulticidal
and ovicidal activities against adults and eggs of T.
urticae (Choi et al. 2004). Very little information, how-
ever, exists in relation to the acaricidal activity of
essential oils against acaricide-resistant T. urticae and
its predatory mite, Neoseiulus californicus McGregor
(Acari: Phytoseiidae).
In this article, we assess the potential of plant es-
sential oils for use as future acaricides. Fumigant ac-
tivity of 10 essential oils against adults of an acaricide-
susceptible strain and four strains of T. urticae resistant
to chlorfenapyr, fenpropathrin, pyridaben, and abam-
ectin was assessed using vapor-phase mortality bioas-
say because the adulticidal activity of these oils was
largely a result of vapor action (Choi et al. 2004). Also,
toxic effects of the essential oils on N. californicus were
examined because of its potential as a predator of T.
urticae in apple (Malus spp.) orchards (Jung et al. 2006).
Materials and Methods
Materials. Ten essential oils used in this study were
as follows: basil (Ocimum basilicum L.), caraway (Ca-
rum carvi L.) seed, citronella Java (Cymbopogon nar-
dus L.), clove (Eugenia caryophyllata Thunberg) leaf,
lemon eucalyptus (Eucalyptus citriodora Hook), pen-
nyroyal (Mentha pulegium L.), peppermint (Mentha
piperita L.), sage (Salvia officinalis L.) Dalmatian,
spearmint (Mentha spicata L.), and thyme (Thymus
vulgaris L.) red purchased from Berjé (BloomÞeld,
NJ; http://www.berjeinc.com). Four commercial
acaricides used in this study were as follows: chlorfe-
napyr 100 g/liter suspension concentrate (SC)
(Dongbu HITek, Seoul, Korea); fenpropathrin 50 g/li-
ter emulsiÞable concentrate (EC) (Dongbang Agro
Corporation, Seoul, Korea); pyridaben 200 g/kg wet-
table powder (WP) (HanKook SamGong, Seoul); and
abamectin 18 g/liter EC (Kyung Nong Co, Seoul). All
other chemicals were reagent grade and available
commercially.
Mites. Cultures of the susceptible T. urticae strain
were maintained in the laboratory without exposure to
any known acaricide from 1992. The chlorfenapyr-
resistant (CRT-53), fenpropathrin-resistant (FRT-
53), pyridaben-resistant (PRT-53), and abamectin-re-
sistant (ART-53) strains had been produced by
selecting a Þeld colony (YA) with chlorfenapyr 100
g/liter SC, fenproxymate 50 g/liter SC, pyridaben 200
g/kg WP, and abamectin 18 g/liter EC, respectively,
for 53 generations at a selection pressure of 30 Ð50%
mortality for successive generations. The YA colony
was originally collected from an apple orchard at
Youngcheon (Kyungbuk Province, Korea) in 2002
that showed resistance to pyridaben (resistance ratio
[RR], 96.7], fenpyroximate (RR, 9.0), dicofol (RR,
8.7), and fenpropathrin (RR, 6.7) but little or no tol-
erance to abamectin (RR, 2.2). The detailed accounts
on the procedures were described previously (Kim et
al. 2004). Mites were reared in acrylic cages (40 by 40
by 55 cm) on kidney bean (Phaseolus vulgaris humilis
Vol. 103, no. 4
Alefeld) seedlings (3 wk after germination) at 24 ⫾
1⬚C, 40 Ð 60% RH, and a photoperiod of 16:8 (L:D) h.
N. californicus, originally collected from citrus or-
chards in Jeju Island in 2006, was maintained in the
laboratory without exposure to any acaricide under
the same rearing conditions. Its host species used was
the susceptible T. urticae reared on kidney bean seed-
lings as described above.
Bioassay. A spray bioassay (Cho et al. 1995) was
used to evaluate the toxicity of four test acaricidesÑ
chlorfenapyr, fenproximate, pyridaben, and abamec-
tinÑagainst adults of the Þve T. urticae strains. Bio-
assays were conducted from early April to mid-April.
In brief, leaf disks (3 cm in diameter) were punched
from leaves of 2-wk-old kidney bean plants. Groups of
20 adult T. urticae (both sexes, 2Ð3 d old) were trans-
ferred onto leaf disks on wet cotton pads in petri
dishes. Four to six concentrations of test chemical
solutions were sprayed at the rate of 0.1 ml per leaf disk
by using a glass spray unit connected to a forced air
supply (PaciÞc Chemical, Seoul, Korea) (Kim et al.
2004). Controls received 0.1 ml of water.
Because the essential oils have fumigant action
(Choi et al. 2004), a vapor-phase mortality bioassay
was used to evaluate the toxicity of oils against adults
of the Þve T. urticae strains. Bioassays were conducted
from mid-April to late June. In brief, leaves (3 wk after
germination) of kidney bean, grown in greenhouse,
were collected and disks (3 cm in diameter) were
punched from each leaf. Each leaf disk with 20 adult
T. urticae (both sexes, 2Ð3 d old) was placed on water-
soaked cotton pads in a petri dish (50 by 10 mm). The
dish was covered using an original tight-Þtting lid with
a Þne wire sieve (4.7 cm in diameter) Þtted over the
central hole (4.5 cm in diameter). Four to six con-
centrations (6.4 Ð159.2 ␮g/cm3) of each test essential
oil dissolved in 80 ␮l of ethanol were applied to What-
man no. 2 Þlter papers (5 cm in diameter; Whatman,
Maidstone, United Kingdom). Control Þlter papers
received 80 ␮l of ethanol. After drying in a fume hood
for 2 min, each Þlter paper was placed over the wire
sieve. This method prevented direct contact of test
adults with the test essential oil. Each dish was then
sealed with another solid lid.
Using the method mentioned above, groups of 20
female N. californicus (2Ð3 d old) were placed on 3-cm-
diameter leaf disks. Treated and control (ethanol or
water only) adults were then held under the same con-
ditions used for colony maintenance. Adult mortalities
were determined at 24 h posttreatment under a binoc-
ular microscope (20⫻). Mites were considered to be
dead if their bodies or appendages did not move when
prodded with Þne wooden dowels (Kim et al. 2004). All
treatments were replicated Þve times using 20 mites per
replicate. Because not all bioassays could be conducted
at the same time, treatments were blocked over time
with a separate control treatment included in each block.
Fleshly prepared solutions were used for each block of
bioassays (Robertson and Preisler 1992).
Data Analysis. Data were corrected for control mor-
tality using the formula of Abbott (1925). Concentra-
tionÐmortality data were subjected to probit analysis
August 2010 HAN ET AL.: TOXICITY OF ESSENTIAL OILS TO T. urticae AND N. californicus 1295

Table 1. Twenty-four hour toxicity of four acaricides to adults from the susceptible (S), CRT-53, FRT-53, PRT-53, and ART-53
strains of T. urticae via spraying

Strain Acaricide na Slope ⫾ SE LC50 (95% CL) mg/liter ␹2 P RR

S Chlorfenapyr 540 1.6 ⫾ 0.14 0.35 (0.28⫺0.42) 6.18 0.06
Fenpropathrin 540 1.2 ⫾ 0.35 2.69 (1.30⫺5.57) 6.62 0.13
Pyridaben 540 0.8 ⫾ 0.11 16.53 (11.25⫺22.86) 7.63 0.18
Abamectin 540 1.9 ⫾ 0.54 0.017 (0.007⫺0.040) 4.10 0.25
CRT-53 Chlorfenapyr 450 ⬎3,200 ⬎9,142
FRT-53 Fenpropathrin 450 1.8 ⫾ 0.17 251.81 (152.11⫺367.13) 6.97 0.73 94
PRT-53 Pyridaben 450 ⬎6,400 ⬎387
ART-53 Abamectin 450 1.4 ⫾ 0.10 1.44 (0.92⫺1.87) 4.46 0.47 85

a
Total number of mites tested; four to six concentrations with Þve replicates per concentration and 20 mites per replicate.

(SAS Institute 2004). The LC50 values for each species
and their treatments were considered to be signiÞ-
cantly different from one another when 95% CL of the
LC50 values failed to overlap. A resistance ratio (RR)
was calculated according to the formula, RR ⫽ LC50
of adults from the acaricide-resistant strain of T. ur-
ticae/LC50 of adults from susceptible strain. RR values
of ⬍10, 10 Ð 40, 40 Ð160, and ⬎160 were classiÞed as
low, moderate, high, and extremely high levels of
resistance, respectively (Kim et al. 2004). Selective
toxicity ratio (STR) was calculated as the ratio of N.
californicus LC50/T. urticae LC50, as reported previ-
ously (Yi et al. 2006).
Results
Resistance. The comparative toxicity of four acari-
cides to the susceptible, CRT-53, FRT-53, PRT-53, and
ART-53 adults was examined by the spray method
(Table 1). Based on 24-h LC50 values, CRT-53 and
PRT-53 adults exhibited extremely high level of resis-
tance to chlorfenapyr (RR, ⬎9,140) and pyridaben
(RR, ⬎390), respectively. FRT-53 and ART-53 adults
were highly resistant to fenpropathrin (RR, 94) and
abamectin (RR, 85), respectively. Mortality for water-
treated controls was ⬍3%.
Susceptibility. The toxicity of 10 essential oils
against the susceptible adults was evaluated by the
vapor-phase mortality bioassay (Table 2). Based on
24-h LC50 values, lemon eucalyptus (19.3 ␮g/cm3)
was the most toxic oil, followed by peppermint,
citronella Java, thyme red, caraway seed, clove leaf,
and pennyroyal oils (LC50, 20.6 Ð23.7 ␮g/cm3). How-
ever, there were no signiÞcant differences in toxicities
among them. Spearmint and basil oils were signiÞ-
cantly less toxic. Sage Dalmatian oil was the least toxic.
Mortality for ethanol-treated controls was ⬍2%.
To female N. californicus, lemon eucalyptus (LC50,
21.4 ␮g/cm3) was likewise the most toxic oil, followed
by citronella Java, thyme red, and pennyroyal oils
(22.5Ð24.9 ␮g/cm3), but no signiÞcant differences in
toxicities among them was observed (Table 2). Pep-
permint, clove leaf, caraway seed, spearmint, and sage
Dalmatian oils were signiÞcantly less toxic. Basil oil
was the least toxic.
The STR was dependent on essential oil used (Table
2). There was no great difference in toxicity of the
other eight oils between them (STR ⫽ 1.0 Ð1.4), al-
though N. californicus was ⬇two-fold more tolerant to
basil and caraway seed oils than T. urticae.
Toxicity to Resistant Mites. The toxicity of 10 es-
sential oils to CRT-53 adults was assessed using the
vapor-phase mortality bioassay (Table 3). Pennyroyal
oil (LC50, 12.1 ␮g/cm3) was signiÞcantly more toxic
than lemon eucalyptus, thyme red, citronella Java,
peppermint, clove leaf, and caraway seed oils (LC50,
20.6 Ð33.4 ␮g/cm3). Spearmint and basil oils were sig-
niÞcantly less active. Sage Dalmatian oil was the least
toxic. Lemon eucalyptus, thyme red, citronella Java,
peppermint, clove leaf, and spearmint oils were of

Table 2. Twenty-four hour fumigant toxicity of 10 test essential oils to susceptible adult T. urticae and female N. californicus

T. urticae N. californicus
Essential oil Slope LC50 (95% CL) Slope LC50 (95% CL) STRb
na ␹2 P na ␹2 P
(⫾ SE) ␮g/cm3 (⫾ SE) ␮g/cm3
Lemon eucalyptus 300 2.3 ⫾ 0.40 19.3 (16.3Ð25.2)a 4.18 0.99 300 4.2 ⫾ 0.45 21.4 (19.6Ð23.4)a 11.67 0.56 1.1
Caraway seed 400 2.6 ⫾ 0.24 22.4 (19.7Ð25.7)a 17.52 0.49 300 5.6 ⫾ 0.51 45.0 (41.7Ð48.4)c 18.08 0.15 2.0
Citronella Java 300 4.5 ⫾ 0.46 22.5 (20.6Ð24.4)a 17.80 0.17 300 5.0 ⫾ 0.53 22.5 (20.8Ð24.2)a 4.76 0.98 1.0
Thyme red 300 6.1 ⫾ 0.74 22.7 (21.1Ð24.1)a 6.38 0.93 300 4.3 ⫾ 0.45 23.2 (21.2Ð25.3)a 9.24 0.75 1.0
Peppermint 400 2.3 ⫾ 0.26 22.8 (19.8Ð27.0)a 21.75 0.19 400 3.7 ⫾ 0.35 28.6 (26.2Ð31.2)ab 7.52 0.98 1.3
Clove leaf 300 4.6 ⫾ 0.43 23.6 (21.6Ð25.8)a 6.56 0.92 300 3.8 ⫾ 0.38 32.7 (29.6Ð36.2)b 11.60 0.56 1.4
Pennyroyal 300 3.5 ⫾ 0.36 23.7 (21.2Ð26.4)a 11.85 0.54 400 4.2 ⫾ 0.36 24.9 (22.8Ð27.0)a 4.04 0.99 1.0
Spearmint 400 2.7 ⫾ 0.26 38.8 (34.5Ð44.0)b 22.90 0.19 300 5.2 ⫾ 0.49 45.5 (42.1Ð49.1)c 10.88 0.62 1.2
Basil 300 4.7 ⫾ 0.46 39.5 (36.1Ð42.9)b 2.49 0.99 300 8.4 ⫾ 0.79 72.6 (69.1Ð76.1)e 8.78 0.79 1.8
Sage Dalmatian 300 2.9 ⫾ 0.34 63.7 (56.2Ð72.3)c 9.81 0.71 300 4.5 ⫾ 0.41 62.0 (56.6Ð67.9)d 7.32 0.88 1.0

Means within a column followed by the same letter are not signiÞcantly different based on nonoverlap of conÞdence limits of the LC50.
a
Total number of mites tested; four to six concentrations with Þve replicates per concentration and 20 mites per replicate.
b
Selective toxicity ratio, N. californicus LC50/T. urticae LC50.
1296 JOURNAL OF ECONOMIC ENTOMOLOGY Vol. 103, no. 4

Table 3. Twenty-four hour fumigant toxicity of 10 test essential oils to chlorpenapyr-resistant CRT-53 adults of T. urticae

Essential oil na Slope ⫾ SE LC50 (95% CL) ␮g/cm3 ␹2 P

Pennyroyal 300 4.7 ⫾ 0.44 12.1 (11.1Ð13.2)a 14.72 0.32
Lemon eucalyptus 300 5.7 ⫾ 0.51 20.6 (19.1Ð22.1)b 7.32 0.89
Thyme red 300 4.8 ⫾ 0.64 22.8 (20.9Ð24.6)b 2.46 0.99
Citronella Java 400 5.4 ⫾ 0.69 23.4 (21.7Ð25.0)b 1.43 0.23
Peppermint 300 4.6 ⫾ 0.47 24.4 (22.4Ð26.5)b 11.23 0.59
Clove leaf 300 3.6 ⫾ 0.37 26.5 (23.9Ð29.5)b 6.46 0.93
Caraway seed 300 2.9 ⫾ 0.30 33.4 (28.6Ð38.3)b 16.46 0.17
Spearmint 300 5.3 ⫾ 0.49 42.2 (39.0Ð45.5)c 9.17 0.76
Basil 300 4.9 ⫾ 0.48 47.2 (43.5Ð51.0)c 14.85 0.32
Sage Dalmatian 300 6.0 ⫾ 0.54 94.7 (87.9Ð101.3)d 14.24 0.36

Means within a column followed by the same letter are not signiÞcantly different based on non-overlap of conÞdence limits of the LC50.
a
Total number of mites tested; four to six concentrations with Þve replicates per concentration and 20 mites per replicate.

equal toxicity against both the S and CRT-53 adults,
indicating a lack of cross-resistance in the CRT-53.
There were signiÞcant differences in toxicities of pen-
nyroyal, caraway seed, basil, and sage Dalmatian oils
between the S and CRT-53 adults.
Against FRT-53 adults (Table 4), lemon eucalyptus
and pennyroyal oils (LC50, ⬇18.4 ␮g/cm3) were sig-
niÞcantly more toxic than citronella Java, thyme red,
peppermint, and clove leaf oils (LC50, 22.3Ð24.4 ␮g/
cm3). The other four oils were signiÞcantly less toxic.
With the exceptions of pennyroyal and caraway seed
oils, the toxicity of the other eight oils was almost
identical against adults from either of the two strains,
indicating a lack of cross-resistance in the FRT-53.
Against PRT-53 adults (Table 5), pennyroyal oil
(LC50, 16.6 ␮g/cm3) was signiÞcantly more toxic than
lemon eucalyptus, thyme red, citronella Java, clove
leaf, and peppermint oils (LC50, 20.8 Ð23.9 ␮g/cm3).
The other four oils were signiÞcantly less effective.
The other eight oils are virtually equal in toxicity to
both S and PRT-53 adults except for pennyroyal and
caraway seed oils, indicating a lack of cross-resistance
in the PRT-53.
Against ART-53 adults (Table 6), lemon eucalyptus
(LC50, 18.7 ␮g/cm3) was the most toxic oil, followed
by pennyroyal, citronella Java, clove leaf, thyme red,
and peppermint oils (LC50, 20.3Ð26.9 ␮g/cm3), al-
though there were no signiÞcant toxicities among
them. The other four essential oils were signiÞcantly
less effective. The toxicity of the other seven oils was
almost identical against adults from either of the two
strains except for caraway seed, spearmint, and sage
Dalmatian oils, indicating a lack of cross-resistance in
the FRT-53.
Discussion
Essential oils consist of highly complex mixtures of
hydrocarbons, such as terpenes (monoterpenes, ses-
quiterpenes, and diterpenes), and oxygenated com-
pounds, such as esters, aldehydes, ketones, alcohols,
phenols, and oxides (Lawless 2002, Isman 2006). They
jointly or independently contribute to behavioral ef-
Þcacy, such as repellence and feeding deterrence, and
physiological efÞcacy, such as acute toxicity and de-
velopmental disruption, against various arthropod
species (Singh and Upadhyay 1993, Ahn et al. 2006,
Isman 2006). Very little work has been done to con-
sider their potential to manage acaricide-resistant T.
urticae. In the current study, citronella Java, clove leaf,
lemon eucalyptus, pennyroyal, peppermint, and
thyme red essential oils exhibited toxicity against adult
T. urticae and showed some potential as future control
agents. These Þndings support earlier work illustrating
certain essential oils may hold promise as novel and
effective acaricidal products against Þeld populations
of T. urticae (Isman 2001, Miresmailli and Isman 2006).
Some products based on plant essential oils, such as
cinnamon, eucalyptus and garlic oils have been com-
mercialized (Isman 2000, 2006). Overall, there are few
botanical acaricides because there are some formida-
ble barriers to commercialization (Isman 2006).

Table 4. Twenty-four hour fumigant toxicity of 10 test essential oils to fenpropathrin-resistant FRT-53 adults of T. urticae

Essential oil na Slope ⫾ SE LC50 (95% CL) ␮g/cm3 ␹2 P

Lemon eucalyptus 300 7.9 ⫾ 0.77 18.4 (17.5Ð19.3)a 14.12 0.37
Pennyroyal 300 6.0 ⫾ 0.59 18.5 (17.1Ð20.0)a 12.83 0.46
Citronella Java 300 4.1 ⫾ 0.49 23.3 (21.2Ð25.8)b 16.08 0.14
Thyme red 300 5.3 ⫾ 0.68 23.5 (21.7Ð25.1)b 3.85 0.99
Peppermint 300 5.0 ⫾ 0.48 23.5 (21.7Ð25.4)b 9.87 0.70
Clove leaf 300 4.7 ⫾ 0.44 24.4 (22.4Ð26.7)b 16.89 0.20
Basil 300 4.7 ⫾ 0.46 42.8 (39.2Ð46.3)c 5.43 0.96
Spearmint 300 5.4 ⫾ 0.50 44.8 (41.5Ð48.2)c 13.19 0.43
Caraway seed 300 5.0 ⫾ 0.48 46.2 (42.6Ð49.9)c 12.16 0.51
Sage Dalmatian 300 3.3 ⫾ 0.38 56.5 (50.2Ð63.6)d 12.10 0.44

Means within a column followed by the same letter are not signiÞcantly different based on nonoverlap of conÞdence limits of the LC50.
a
Total number of mites tested; four to six concentrations with Þve replicates per concentration and 20 mites per replicate.
August 2010 HAN ET AL.: TOXICITY OF ESSENTIAL OILS TO T. urticae AND N. californicus 1297

Table 5. Twenty-four hour fumigant toxicity of 10 test essential oils to pyridaben-resistant PRT-53 adults of T. urticae

Essential oil na Slope ⫾ SE LC50 (95% CL) ␮g/cm3 ␹2 P

Pennyroyal 300 7.3 ⫾ 0.74 16.6 (15.7Ð17.5)a 3.30 0.99
Lemon eucalyptus 300 5.8 ⫾ 0.52 20.8 (19.2Ð22.3)b 7.71 0.86
Thyme red 400 5.7 ⫾ 0.50 21.5 (20.2Ð22.8)b 7.48 0.98
Citronella Java 300 4.9 ⫾ 0.52 22.0 (20.3Ð23.7)b 5.53 0.96
Clove leaf 300 4.0 ⫾ 0.39 23.2 (20.9Ð25.5)b 5.07 0.97
Peppermint 300 5.4 ⫾ 0.51 23.9 (22.1Ð25.7)b 18.66 0.13
Basil 300 5.9 ⫾ 0.53 40.9 (37.9Ð43.9)c 7.95 0.85
Spearmint 300 5.0 ⫾ 0.48 44.5 (41.0Ð48.1)c 16.88 0.20
Caraway seed 300 4.2 ⫾ 0.47 44.1 (40.2Ð48.3)c 16.74 0.16
Sage Dalmatian 300 4.3 ⫾ 0.40 61.7 (56.2Ð67.8)d 17.28 0.19

Means within a column followed by the same letter are not signiÞcantly different based on nonoverlap of conÞdence limits of the LC50.
a
Total number of mites tested; four to six concentrations with Þve replicates per concentration and 20 mites per replicate.

Investigations of the modes of action and resistance
mechanisms of natural acaricidal products and acari-
cides are of practical importance for T. urticae control
because they may give useful information on the most
appropriate formulations and delivery means and for
future resistance management. Mechanisms of resis-
tance to acaricides include reduced penetration, tar-
get site insensitivity (altered acetylcholinesterse), and
enhanced metabolism (Schoknecht and Otto 1992,
Knowles 1997, Devine et al. 2001, Nauen et al. 2001).
Alternative control products with novel modes of ac-
tion, low mammalian toxicity, and little environmental
impact are badly needed. Certain plant preparations
or their constituents are highly effective against in-
secticide-resistant insect pests (Ahn et al. 1997). Our
current Þndings that some essential oils are virtually
equal in toxicity to both acaricide-susceptible strain
and four strains resistant to chlorfenapyr, fenproxi-
mate, pyridaben, and abamectin suggest that these
essential oils and the acaricides do not share a common
mode of action or elicit cross-resistance. In addition,
fumigant action of essential oils as demonstrated
through our vapor-phase mortality bioassay is of prac-
tical importance because volatile oil compounds could
penetrate plant canopies resulting in enhanced efÞ-
cacy. Although not yet proven, octopaminergic and
␥-aminobutyric acid receptors have been suggested as
novel target sites for some essential oil constituents by
Kostyukovsky et al. (2002) and Priestley et al. (2003),
respectively.
The uses of acaricides and natural enemies have
become key components of IPM in fruit orchards and
greenhouses. Unfortunately, these two practices are
not always compatible because predatory mites are
often more susceptible to many of the commonly used
acaricides than the phytophagous target mites (Kim et
al. 1999, Ahn et al. 2001, Choi et al. 2004). N. califor-
nicus is one of the most promising biological control
agents of T. urticae since it was Þrst identiÞed in col-
lections made from citrus orchards in Jeju Island in
2006 (Jung et al. 2006). In the current study, the 10
essential oils tested were also toxic to female N. cali-
fornicus. There were no signiÞcant differences in tox-
icities between T. urticae and N. californicus with the
exceptions of caraway seed, clove leaf, and basil oils. Our
results suggest that if N. californicus is used as a biological
control agent, its release time should be properly se-
lected to minimize fumigant toxicity of the oils.
In conclusion, the essential oils described could be
useful as fumigants in the control of T. urticae popu-
lations in greenhouses, particularly due to their activ-
ity against acaricide-resistant T. urticae, provided that
a carrier giving a slow release of active material can be
selected or developed and leakage out of greenhouses
is minimized. For the practical use of the essential oils
as novel acaricides to proceed, further research is
needed to establish their human safety. However, es-
sential oils are commonly used as fragrances and ßa-
voring agents for foods, beverages, and cosmetics (Is-
man 2000, Lawless 2002). In addition, their acaricide
modes of action and changes in the quality of crops
treated with the oils (e.g., color, ßavor, odor, and
texture) need to be established and formulations (e.g.,
fumigant and smoking agent) for improving acaricidal

Table 6. Twenty-four hour fumigant toxicity of 10 test essential oils to abamectin-resistant ART-53 adults of T. urticae

Essential oil na Slope ⫾ SE LC50 (95% CL) ␮g/cm3 ␹2 P

Lemon eucalyptus 300 5.0 ⫾ 0.51 18.7 (17.3Ð20.2)a 15.78 0.26
Pennyroyal 300 5.2 ⫾ 0.48 20.3 (18.7Ð21.9)a 3.83 0.99
Citronella Java 300 5.3 ⫾ 0.54 21.6 (20.0Ð23.2)a 19.78 0.10
Clove leaf 300 4.4 ⫾ 0.42 23.5 (21.5Ð25.8)a 12.31 0.50
Thyme red 300 7.0 ⫾ 0.72 26.0 (24.6Ð27.5)a 5.66 0.96
Peppermint 300 4.4 ⫾ 0.47 26.9 (24.7Ð29.4)a 9.85 0.71
Caraway seed 300 5.1 ⫾ 0.49 48.0 (44.4Ð51.8)b 16.04 0.25
Spearmint 300 4.0 ⫾ 0.44 49.0 (44.6Ð53.9)b 11.55 0.56
Sage Dalmatian 300 4.6 ⫾ 0.42 63.7 (58.3Ð69.7)c 13.66 0.40
Basil 300 9.2 ⫾ 0.85 65.3 (62.2Ð68.3)c 2.69 0.99

Means within a column followed by the same letter are not signiÞcantly different based on nonoverlap of conÞdence limits of the LC50.
a
Total number of mites tested; four to six concentrations with Þve replicates per concentration and 20 mites per replicate.
1298 JOURNAL OF ECONOMIC ENTOMOLOGY
potency and stability, thereby reducing costs, need to
be developed.
Acknowledgments
This work was carried out with the support of Cooperative
Research Program for Agriculture Science & Technology
Development (PJ006859), Rural Development Administra-
tion; and the Ministry of Education, Science and Technology
of the Korean Government (World Class University Program
R31-10056) (to Y.-J.A.).
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Received 9 July 2009; accepted 7 April 2010.