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Insecticidal and Acetylcholine Esterase Inhibition Activity of Asteraceae Plant Essential Oils and Their Constituents against Adults of the German Cockroach (Blattella germanica) Hwa-Jeong Yeom, Chan-Sik Jung, Jaesoon Kang, Junheon Kim, Jae-Hyeon Lee, Dong-Soo Kim, Hyun-Seok Kim, Pil-Sun Park, Kyu-Suk Kang, and Il-Kwon Park J. Agric. Food Chem., Just Accepted Manuscript • DOI: 10.1021/jf505927n • Publication Date (Web): 09 Feb 2015 Downloaded from http://pubs.acs.org on February 16, 2015

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Journal of Agricultural and Food Chemistry

Insecticidal and Acetylcholine Esterase Inhibition Activity of

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Asteraceae Plant Essential Oils and Their Constituents against Adults of

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the German Cockroach (Blattella germanica)

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Hwa-Jeong Yeom1†, Chan-Sik Jung1†, Jaesoon Kang2, Junheon Kim3, Jae-

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Hyeon Lee4, Dong-Soo Kim5, Hyun-Seok Kim6,7, Pil-Sun Park6,7, Kyu-Suk

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Kang6,7 and Il-Kwon Park6,7* 1

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Division of Forest Insect Pests and Diseases, Korea Forest

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Research Institute, Seoul 130-712, Republic of Korea; 2Gyeongnam Department of

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Environmental Toxicology and Chemistry, Korea Institute of Toxicology, Jin-Ju,

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Gyeongnam, Republic of Korea

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3

Institute of Agriculture and Life Science/BK21+, Gyeongsang National University, Jinju,

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Gyeongnam 660-701, Republic of Korea; 4Department of Seed & Seedling Management,

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Korea Forest Seed and Variety Center, Chungcheongbuk-do 380-941, Republic of Korea; 5

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Southern Forest Research Center of the Korea Forest Research Institute, Jinju,

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Gyeongnam 660-300, Republic of Korea; 6Department of Forest Science, 7Research

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Institute of Agriculture and Life Science, College of Agriculture and Life Sciences, Seoul

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National University, Seoul 151-921, Republic of Korea

18

†

19

*

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873-3560; e-mail [email protected]).

These authors contributed equally to this work To whom correspondence should be addressed (telephone +82-2-880-4751; fax +82-2-

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ACS Paragon Plus Environment


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ABSTRACT

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The fumigant and contact toxicities of 16 Asteraceae plant essential oils and their

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constituents against adult male and female Blattella germanica were examined. In a

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fumigant toxicity test, tarragon oil exhibited 100% and 90% fumigant toxicity against adult

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male German cockroaches at 5 and 2.5 mg/filter paper, respectively. Fumigant toxicities of

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Artemisia arborescens and santolina oils against adult male German cockroaches were 100%

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at 20 mg/filter paper, but were reduced to 60% and 22.5% at 10 mg/filter paper,

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respectively. In contact toxicity tests, tarragon and santolina oils showed potent insecticidal

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activity against adult male German cockroaches. Components of active oils were analyzed

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using gas chromatography, gas chromatography-mass spectrometry or nuclear magnetic

31

resonance spectrometer. Among the identified compounds from active essential oils,

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estragole demonstrated potent fumigant and contact toxicity against adult German

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cockroaches. β-Phellandrene exhibited inhibition of male and female German cockroach

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acetylcholinesterase activity with IC50 values of 0.30 and 0.28 mg/mL, respectively.

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Key words: Asteraceae plant essential oils; fumigant toxicity; contact toxicity; German

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cockroach; acetylcholinesterase inhibition

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INTRODUCTION

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The German cockroach, Blattella germanica (L.) is the most common cockroach found

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in human residences. Cockroaches are considered a significant insect pest with regard to

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hygiene, because not only are they a source of allergy,1 but they also spread several

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intestinal diseases such as cholera, diarrhea, and dysentery.2 To control German

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cockroaches, several contact or residual insecticides such as organophosphorus, carbamate,

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pyrethroids, and hydrmethylnon have been used,3-5 but these pesticides have many side

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effects, including environmental and human health problems, and are susceptible to

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resistance.4-6 Because of multiple side effects caused by synthetic pesticides, the

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development of environmentally friendly German cockroach control agents is essential.7-9

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Plant essential oils extracted by steam distillation are good resources for developing

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German cockroach control agents because they are known to have many bioactivities

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including insecticidal and repellent activity against German cockroaches.8-13 Another

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advantage of plant essential oils is their high volatility. The main constituents of essential

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oils are mono- or sesqui- terpenoids, which are highly volatile.14-15 High volatility reduces

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concerns of residue problems. Furthermore, constituents of some plant essential oils are

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known to act synergistically with respect to insecticidal activity,8-9 which could retard the

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development of resistance if the oil constituents have different modes of action.

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In this study, we investigated the fumigant and contact toxicities of Asteraceae plant

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essential oils and their constituents against male and female adult German cockroaches to

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find alternatives to current insecticides. We also measured the acetylcholinesterase (AChE)

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inhibition activities of constituents from active Asteraceae plant essential oils to better

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understand their mode of action.

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MATERIALS AND METHODS

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Plant Essential Oils and Chemicals. Essential oils of Artemisia afra and davana were

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purchased from Jinarome (Anyang, Gyeonggi Province, Korea). Artemisia arborescens,

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chamomile blue, chamomile roman, chamomile wild, chrysantheme abs., costus root,

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elecampane roots, erigeron, Eriocephalus punctulatus, Helichrysum bracteiferum,

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helichrysum (Immortella), santolina, tagetes extra-s, and tarragon were purchased from

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Oshadhi (Weinstrasse, Bühl/Baden, Germany). These plant essential oils are listed in Table

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1. Camphene (80%), estragole (98%), (+)-limonene (97%), myrcene (95%) and cis-ocimene

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(90%) were purchased from Sigma-Aldrich (Milwaukee, WI, USA). Camphor (>97%),

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terpinen-4-ol (99%), γ-terpinene (97%), and p-cymene (95%) was purchased from Fluka

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(Buchs, Switzerland). β-Caryophyllene (>90%), (+)-α-pinene (95%), and β-pinene (94%)

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were obtained from Tokyo Kasei (Tokyo, Japan). Acetone (99.8%) was purchased

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from Merck. Dichlorvos (purity 98.6%) was purchased from Chem Service (West Chester,

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PA, USA) and deltamethrin (purity 96.8%) was supplied from Dongbu Farm Hannong

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(Seoul, Republic of Korea).

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Insects. German cockroaches, B. germanica, were reared in plastic boxes

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(54.3×18.8×36.3 cm, L×W×H) without exposure to any insecticide. The cockroaches were

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supplied with water from a glass flask fitted with a cotton stopper and dried mouse food

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(Purina feed). The cockroaches were maintained at 25 ± 1 °C and 60% RH (relative

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humidity) under a 16:8 h light:dark cycle.

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Gas Chromatography. Gas chromatography (GC) analysis of Artemisia arborescens,

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santolina, and tarragon oils was carried out using an Agilent 7890A (Agilent Technologies,

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Santa Clara, CA, USA), equipped with DB-1MS or HP-INNOWAX columns (internal

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diameter [ID], 30 m × 0.25 mm; film thickness, 0.25 µm; J&W Scientific, Folsom, CA,

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USA) and a flame ionization detector (FID). Retention times of essential oil components

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were compared with those of authentic compounds. The oven temperature of the gas

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chromatograph was programmed as isothermal at 40 °C for 1 min, raised at a rate of

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6 °C/min to 250 °C, and maintained at 250 °C for 4 min. Helium was used as the carrier

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gas at a flow rate of 1.5 mL/min. Retention indices were obtained in relation to a

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homologous series of n-alkanes (C7-C20; DB-1MS), (C8-C22; HP-INNOWAX) under the

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same gas chromatography operating conditions. Further identification of oil constituents

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was confirmed by increasing the integrated area by co-injection with oil and standard

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samples.

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Gas Chromatography-Mass Spectrometry. Constituents of Asteraceae plant essential

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oils were also analyzed using a tandem gas chromatograph (Agilent 7890A)-mass

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spectrometer (Agilent 5975C MSD) (GC-MS) (Agilent Technologies). A DB-5MS column

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(ID, 30 m × 0.25 mm; film thickness, 0.25 µm; J&W Scientific) was used, and the column

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oven temperature program was the same as used for GC-FID analysis. Helium was used as

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the carrier gas at a flow rate of 1.0 mL/min. The effluent of the column was introduced

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directly into the source of the MS via a transfer line (250 °C). Ionization was attained using

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electron impact (70 eV; source temperature, 230 °C). The scan range was 41-400 amu.

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Most components of the plant essential oils were tentatively confirmed by comparing the

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mass spectra of each peak with those of standard samples in the NIST MS library. NMR

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spectra were obtained on a Varian UI 500 NMR spectrometer (500 MHz for 1H spectra and

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125 MHz for

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standard.

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13

C spectra) at Korean Basic Science Institute using TMS as an internal

Isolation and Identification of ß-Thujone and Artemisia Ketone. To isolate β-thujone,

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1 mL of A. arborescens oil was chromatographed on a SiO2 column (Wakogel® 200; Wako

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Pure Chemicals, Osaka, Japan) eluting with a diethyl ether-hexane gradient. β-Thujone and

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camphor

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rechromatographed on a 10% AgNO3-impregnated SiO2 column, and β-thujone eluted with

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10% ether-hexane solution in 98.9% purity (395 mg). Artemisia ketone was isolated from

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santolina oil. Santolina oil (1 mL) was chromatographed on a SiO2 column eluting with a

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diethyl ether-hexane gradient. Artemisia ketone (purity 80%) was isolated with 10:90

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(ether: hexane) solution. To obtain pure artemisia ketone, the 10% ether fraction was-

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rechromtographed on a SiO2 column eluting with 1:99 ether: hexane. The purity of isolated

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artemisia ketone (245 mg) was 98.6%. Isolated β-thujone and artemisia ketone were

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subjected to NMR analysis and were used for bioassays.

coeluted

with

10:90

(ether:

hexane)

solution.

This

mixture

was

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Synthesis of β-Phellandrene. β-Phellandrene (purity 98.2%) was synthesized in the

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laboratory and was subjected to bioassay. The synthesis procedure and NMR data have

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been well documented in our previous study.16

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Fumigant Toxicity Test. The fumigant toxicity of Asteraceae plant essential oils and

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their constituents was determined using a glass cylinder (diameter, 9.5 cm; height, 19 cm)

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with a wire sieve installed 9.5 cm above the bottom. Asteraceae plant essential oils or their

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components were applied to a paper disc (8 mm, Advantec). The treated paper disc was

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transferred to the bottom lid of the glass cylinder. The lid of the glass cylinder was sealed

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with Para-film to prevent the leakage of the oil or constituents (Pechiney Plastic Packaging

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Company, Chicago, IL, USA). Ten adult male or female German cockroaches were placed

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on the sieve, which prevented direct contact of the cockroaches with the test oils and

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constituents. Test cockroaches were maintained at 25 ± 1 °C and 60% RH. Mortality was

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determined 48 h after treatment. All treatments were replicated 4 times.

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Contact Toxicity Test. The appropriate dose of Asteraceae plant essential oil or

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individual constituents was dissolved in acetone. Adult male and female German

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cockroaches were anesthetized using CO2, and the oils or constituents in acetone were

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topically applied to the abdomen of adult male and female German cockroaches with a

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micro-applicator (Burkard, UK). Control insects received only acetone (1 µL). Treated

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adults (10 adults/ replication) were transferred to petri-dishes (diameter, 9.5 cm; height, 2

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cm) that were maintained at 25 ± 1 °C and 60% RH. Mortality was determined 24 h after

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treatment. Each assay was replicated 5 times.

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Acetylcholinesterase Inhibition. Crude protein was obtained from two adult male and

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female cockroaches. Adult cockroaches were soaked in 0.1 M Tris-HCl mixed with 0.02 M

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NaCl, 0.5% Triton X-100 (pH 7.8), and a protease inhibitor cocktail (Sigma-Aldrich, St.

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Louis, MO, USA), and then they were ground using a glass tissue grinder (Wheaton

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Industries Inc., Millville, NJ, USA) in ice. The ground cockroaches extract was centrifuged

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at 17,000 g for 15 min at 4 °C to eliminate insect tissue debris. The concentration of crude

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protein was measured by the Bradford protein assay using BSA as the standard protein.

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The AChE inhibition activity was evaluated using the modified Ellman method.17 Test

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chemicals were dissolved in acetone to give a concentration of 100 mg/mL. Mixtures of 1

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µl of test compound and 79 µl of crude cockroach protein were placed in 96-well

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microplates, which were incubated for 10 minutes at RT (room temperature). The control

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received acetone only. 10 µl of acetylthiocholine iodide (ASChI, 10 mM) and 10 µl of 5,5'-

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dithiobis-(2-nitrobenzoic acid) (DTNB, 4 mM) were added to the mixtures of test

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compounds and crude protein. AChE activity was evaluated by estimating the initial

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velocity (Vo) for 20 minutes at 30 sec -intervals at 405 nm and RT using an iMark

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microplate absorbance reader (Bio-Rad, Hercules, CA, USA). The primary AChE

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inhibition assay was replicated at least 3 times. The percentage inhibition of each chemical

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was calculated according to the following formula:

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Inhibition activity (%) = 100 - [(Vo of chemical treatment/Vo of control treatment) × 100]

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To determine the IC50 value of β-phellandrene, the following concentrations of β-

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phellandrene were used: 1, 0.5, 0.2, 0.1, and 0.05 mg/mL. All treatments were replicated 3

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times at each concentration. The AChE inhibition activity was evaluated according to the

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formula described above, and the IC50 was determined by probit analysis.18 -8-


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Statistical Analysis. Percentage mortality and primary AChE inhibition rates were

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transformed to arcsine square-root values for analysis of variance. Treatment mean values

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were compared and analyzed using Scheffe's test.18 Mean (±SE) values of untransformed

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data are reported.

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RESULTS AND DISCUSSION

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Fumigant and Contact Toxicities of Plant Essential Oils. Fumigant and contact

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toxicities of 16 Asteraceae plant essential oils varied according to oil type and dose (Table

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2). Artemisia arborescens, santolina, and tarragon essential oils exhibited 100% fumigant

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toxicity against male German cockroaches at 20 mg/filter paper. The fumigant toxicity of

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tarragon was 100% at 10 and 5 mg/filter paper, but declined to 90% at 2.5 mg/filter paper.

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Fumigant toxicities of Artemisia arborescens, and santolina were 60% and 22.5% at 10

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mg/filter paper, respectively. Chamomile roman showed 85% mortality at 20 mg/filter

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paper, but declined to 22.5% at 10 mg/filter paper. Other oils showed moderate or weak

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fumigant toxicity against male German cockroaches. In a test with female German

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cockroaches, only tarragon showed potent fumigant toxicity at 20 mg/filter paper.

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Fumigant toxicities of tarragon against female German cockroaches were 100% and 50%

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at concentrations of 10 and 5 mg/filter paper, respectively. Other oils exhibited moderate

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or weak toxicity against female German cockroaches at 20 mg/filter paper. In contact

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toxicity tests, santolina, and tarragon essential oils showed 100% insecticidal activity

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against adult male German cockroaches at 2 mg/♂ (Table 3). Insecticidal activities of

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tarragon and santolina were 100% and 86% at 1 mg/♂, but were reduced to 44% and 12%

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at 0.5 mg/♂, respectively. Helichrysum bracteiferum, helichrysum (immortelle), and

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chamomile wild exhibited 98%, 98% and 96% mortality against male German cockroaches

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at 2 mg/♂, but declined to 30%, 50% and 54% at 1 mg/♂, respectively. Contact toxicities

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of other oils against male German cockroaches were less than 85% at 2 mg/♂. In tests with

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female cockroaches, tarragon and santolina exhibited 100% and 70% mortality at 2 mg/♂,

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respectively, while the other oils exhibited less than 55% mortality at 2 mg/♂. Many plant

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essential oils have been reported to exhibit fumigant or contact toxicity against German

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cockroaches.8-10, 12-13 Yeom et al.8 reported that plant essential oils belonging to Apiaceae

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exhibited potent fumigant and contact toxicity against German cockroaches. Alzogaray et

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al.19 and Yeom et al.9 also investigated the insecticidal activities of plant essential oils

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belonging to Myrtaceae. However, fumigant and contact toxicities of the plant essential

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oils used in this study against German cockroaches have not been reported in any other

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study.

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Chemical Constituents of Plant Essential Oils. The chemical compositions of

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Artemisia arborescens, santolina, and tarragon essential oils are shown in Table 4. The

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most abundant components of Artemisia arborescens were β-thujone (43.72%) followed

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by camphor (22.93%), β-myrcene (7.22%), sabinene (4.21%), terpinen-4-ol (3.03%), β-

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caryophyllene (2.56%), camphene (2.51%), α-pinene (2.33%), γ-terpinene (2.28), and p-

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cymene (2.09%). Artemisia ketone (31.20%), β-phellandrene (25.67%), β-myrcene

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(11.46%), sabinene (8.99%), and β-pinene (8.84%) were detected as the main constituents

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in santolina oil. Estragole (88.75%) was identified as the most abundant compound in

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tarragon oil followed by limonene (4.87%), cis-ocimene (3.02%), and α-pinene (1.46%).

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Chemical analyses of Artemisia arborescens, santolina, and tarragon essential oils have

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been reported previously in other studies.20-22 While the main components of these oils

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were similar to those previous reports, there were, some differences in minor components

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and composition quantities. Beyrouthy et al.20 reported that β-thujone (68.5%),

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chamazulene (12.3%), terpinen-4-ol (1.5%), myrcene (1.1%), linalool, and cis-thuyanol-4-

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ol (1%), α-thujone (0.9%), and sabinene (0.8%) were detected as the main components of

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Artemisia arborescens. Chamazulene, linalool, cis-thuyanol-4-ol, and α-thujone were not

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detected in our study. The difference of some oil components and composition amount

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could be attributed to the origin of Artemisia arborescens. Beyrouthy et al.20 and we used

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Artemisia arborescens essential oils from Lebanon and Moroco, respectively. Artemisia

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ketone, camphor, and β-phellandrene were analyzed as the main compounds in santolina

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oil by Demirci et al.,21 while camphor was not detected in our study. Arabhosseini et al.22

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analyzed the constituents of French tarragon oil and found estragole, methyl eugenol,

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ocimene, and sabinene as the main constituents. In our study, the amount of estragole was

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higher, and methyl eugenol and sabinene were not identified. Galambosi and Peura23

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suggested that constituents of plant essential oils differ widely according to production

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conditions such as harvesting date, storage time, as well as climatic or edaphic factors. Isolation and Identification of ß-Thujone and Artemisia Ketone. Based on 1H- and

231 232

13

C-NMR, DEPT, 1H-1H COSY, and HMQC, each signal of β-thujone was assigned as

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shown in Figure 1. β-thujone. 13C (ppm): 12.49 (C10, -CH3), 14.71 (C6, -CH2), 19.72 (C9,

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-CH3), 19.79 (C8, -CH3), 24.63 (C5, -CH), 27.42 (C1, -C), 32.30, (C7, -CH), 41.73 (C2, -

235

CH2), 45.41 (C4,-CH), 218.51 (C3, -C=O); 1H (ppm): -0.04 (6a, 1H, dd, J=5.5, 4), 0.59 (6b,

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1H, tq, J=7, 2), 1.00 (8, 3H, d, J=7), 1.03 (10, 3H, d, J=7), 1.44 (5, 7, 2H, m), 2.12 (2a, 1H,

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d, J=18.5), 2.54 (2b, 1H, d, J=18.5, 2), 2.71 (4, 1H, m). Chemical shift values of the proton

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at C6 of α-thujone and β-thujone were reported as 0.12 and -0.05 ppm, respectively.24 1H-

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NMR analysis of the isolated thujone showed only a chemical shift value at -0.05 ppm.

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Thus, the isolated thujone was determined to be as β-thujone. NMR data of artemisia

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ketone was assigned in our previous study.25

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Fumigant and Contact Toxicities of Individual Constituents. The fumigant and

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contact toxicities of individual constituents from Artemisia arborescens, santolina, and

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tarragone essential oils are shown in Tables 5 and 6. Fumigant or contact toxicities of α-

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pinene, β-pinene, β-myrcene, p-cymene, limonene, γ-terpinene, and terpinen-4-ol were not

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tested in this study, because their toxicities to German cockroaches have been reported in

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our previous studies.8,9 In a fumigant toxicity test, estragole was the most toxic components

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to adult males. Fumigant toxicities of estragole were 100% and 67.5% at 2.5 and 1.25

249

mg/filter paper, respectively. β-Thujone, β-phellandrene, and cis-ocimene showed 100%,

250

100% and 82.5% fumigant toxicity against adult males at 20 mg/filter paper, but their

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activities were reduced to 90%, 20%, and 35% at 10 mg/filter paper, respectively. Other

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compounds showed weak activity. In tests with adult females, estragole exhibited 100%

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fumigant toxicity at 20 and 10 mg/filter paper, but its toxicity was reduced to 80% and 40%

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at 5 and 2.5 mg/filter paper, respectively. The fumigant toxicity of β-thujone was 100% at

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20 mg/filter paper, but was reduced to 17.5% at 10 mg/filter paper. Other compounds

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showed moderate or weak toxicity at 20 mg/filter paper. In a contact toxicity test, only

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estragole exhibited potent activity against adult males. The contact toxicity of estragole

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was 100% at 1 and 0.5 mg/♂, but was reduced to 34% at 0.25 mg/♂. Artemisia ketone

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produced 74% mortality at 1 mg/♂, but the mortality decreased to 30% at 0.5 mg/♂. Other

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compounds exhibited moderate or weak contact toxicity. In a contact toxicity test with

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female German cockroaches, all test compounds showed moderate or weak toxicity at 1

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mg/♀. However, fumigant or contact toxicity of all test compounds were weaker than

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conventional pesticides such as dichlorvos and deltamethrin (Table 5 and 6). Insecticidal

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activities of phytochemicals from plant essential oils against German cockroaches have

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been reported in several studies.8,9,10,13 Yeom et al.8,9 reported fumigant and contact

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toxicities of constituents from Apiaceae and Myrtaceae plant essential oils. They found

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that carvone, 1,8-cineole, trans-dihydrocarvone, cumminaldehyde, trans-anethole, p-

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cymene, γ-terpinene, carvacrol, thymol, carveol, terpinen-4-ol, terpinolene, and α-terpinene

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demonstrated potent fumigant or contact toxicities against male and female German

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cockroaches. Philips et al.10 and Philips and Appel13 also tested fumigant and contract

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toxicities of phytochemicals from plant essential oils including carvacrol, 1,8-cineole,

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trans-cinnamaldehyde, citronellic acid, eugenol, geraniol, limonene, linalool, menthone, α-

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pinene, β-pinene, and thymol. In our study, estragole showed the most potent fumigant

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toxicity to male and female German cockroaches and contact toxicity to male German

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cockroaches. Estragole is the main constituent of basil oil, and its insecticidal activity

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against fruit flies (Ceratitis capitata, Bactrocera dorsalis, Bactrocera cucurbitae), maize

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weevil (Sitophilus zeamais), azuki bean weevil (Callosobruchus chinensis), rice weevil

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(Sitophilus oryzae), and cigarette beetle (Lasioderma serricorne) have been reported in

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several studies.26,27,28 However, this is the first report on its fumigant and contact toxicity

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against adult German cockroaches. Many studies have identified β-thujone and β-

281

phellandrene as components of plant essential oils.20,21 However there are few reports on

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their insecticidal activity because they are not commercially available.29

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In this study, adult male German cockroaches were more susceptible than adult female

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German cockroaches to plant essential oils and their constituents. The weight of female

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German cockroaches (average weight of 20 females was 104.9 mg) was about twice that of

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the male German cockroaches (54.2 mg). This weight difference may be the main reason

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for the different susceptibility to test oils and their constituents. Lee et al.30 also found that

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weight differences between male and female German cockroaches affected the insecticide

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susceptibility to synthetic insecticides such as organophosphates, carbamates and

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pyrethroids.

291

Primary AChE Inhibition Assay and IC50 Determination. AChE inhibition by artemisia

292

ketone, camphene, camphor, β-caryophyllene, estragole, cis-ocimene, and β-phellandrene

293

were determined, since inhibition activities of the other constituents were reported in our

294

previous study.8,9 The primary inhibition activities of various isolated constituents against

295

German cockroach acetylcholinesterase are summarized in Figure 2. β-Phellandrene gave

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296

the highest inhibition (81.28%), followed by cis-ocimene (44.10%) and estragole (25.04%)

297

(Fig. 2). Inhibition activities of the other compounds were less than 20%. In the primary

298

inhibition assay, only β-phellandrene showed >50% inhibition. IC50 values of β-

299

phellandrene against male and female acetylcholinesterase were 0.30 and 0.28 mg/mL,

300

respectively (Table 7). Insect acetylcholinesterase inhibition activity of many

301

phytochemicals has been investigated in several studies.16,25,31-38 German cockroach AChE

302

inhibition activity of phytochemicals from plant essential oils has also been studied by

303

Yeom et al.8,9 They reported that insecticidal activity of carvacrol and dihydrocarvone

304

correlated with their ability to inhibit German cockroach acetylcholinesterase. However,

305

there was no relationship between AChE inhibition and most other active compounds. In

306

our study, only β-phellandrene showed potent AChE inhibition activity, while most of the

307

other active compounds revealed moderate or weak inhibition activity. Seo et al.25 reported

308

that β-phellandrene exhibited potent AChE inhibition activity against Japanese termites.

309

The results of our study and other research indicate that insecticidal activity of β-

310

phellandrene correlates with its ability to inhibit AChE. Although the mode of action of

311

phytochemicals from plant essential oils has been reported by some research groups to

312

include AChE, a definitive mechanism has yet to be completely established.8,9,16,25,31,39,40

313

Our results indicate that Asteraceae plant essential oils and their components can be

314

used as fumigants or spray-type control agents against German cockroaches. However,

315

further studies including safety of the oils and their constituents to humans and non-target

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316

organisms, formulations, and their modes of action are necessary for practical use of

317

Asteraceae plant essential oils and their constituents as novel cockroach -control agents.

318 319

ACKNOWLEDGEMENT

320

This work was supported by a Research Resettlement Fund for the new faculty of

321

Seoul National University (Project No.: 500-20140209) and a grant from Korea Forest

322

Service (Project No.: S111414L080110) to I.K. Park.

323 324

REFERENCES

325

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Table 1. List of Asteraceae Plant Essential Oils Tested Common name of essential oil

Scientific name

Source

Part

Country of origin

Artemisia afra

Artemisia afra

Jinarome

Flowering plant

South Africa

Artemisia arborescens

Artemisia arborescens

Oshadhi

Flowering plant

Morocco

Chamomile blue

Chamomilla matricaria

Oshadhi

Blossoms

Nepal

Chamomile roman

Anthemis nobilis

Oshadhi

Blossoms

France

Chamomile wild

Ormensis multicaulis

Oshadhi

Blossoms

Morocco

Chrysantheme abs.

Chrysanthemum morifolium

Oshadhi

Leaves

India

Costus root

Saussurea lappa

Oshadhi

Roots

India

Davana

Artemisia pallens

Jinarome

Leaves

India

Elecampane roots

Inula racemosa

Oshadhi

Roots

India

Erigeron

Coniza canadensis

Oshadhi

Flowering plant

Canada

Eriocephalus punctulatus


Oshadhi

Flowering plant

South Africa

Helichrysum bracteiferum

Helichrysum bracteatum

Oshadhi

Blossoms

South Africa

Helichrysum (Immortella)

Helichrysum angustifolium

Oshadhi

Blossoms

Croatia

Santolina

Santolina chamaecyparissus

Oshadhi

Plant

Spain

Tagetes extra-s

Tagetes minuta

Oshadhi

Blossoms

Egypt

Tarragon

Artemisia dracunculus

Oshadhi

Plant

France

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436

Page 24 of 32

Table 2. Fumigant Toxicity of 16 Asteraceae Plant Essential Oils against Adult of German Cockroaches Mortality (%, Mean±S.E., N=40a) Plant essential oils

male b

20 Artemisia afra Artemisia arborescens

2.5

1.25

20

10

5

2.5

2.5 ± 2.5c

17.5 ± 7.5cd

-

-

-

60 ± 12.9b

15.0 ± 6.5b

25 ± 6.5bc

-

-

-

2.5 ± 2.5ef

0d

-

-

-

85.0 ± 5.0ab

22.5 ± 8.5c

25.0 ± 6.5bc

-

-

-

Chamomile wild

57.5 ± 2.5c

10 ± 4.1c

0d

-

-

-

Chrysantheme abs.

7.5 ± 4.8def

0d

-

-

-

0f

0d

-

-

-

Costus root Davana Elecampane roots Erigeron

27.5 ± 7.5de

35 ± 2.9bc

-

-

-

60 ± 4.1bc

0c

42.5 ± 2.5b

0b

-

-

0d

-

-

-

2.5 ± 2.5c

0d

-

-

-

0f


30 ± 4.1d

Helichrysum bracteiferum

5 ± 5.0def

0d

-

-

-

Helichrysum (Immortella)

0f

0d

-

-

-

100a

22.5 ± 9.6c

37.5 ± 4.8bc

2.5 ± 2.5b

-

-

0f

0d

-

-

-

100a

100a

100a

90

15.0 ± 6.5

100a

100a

50 ± 14.1

10 ± 8.2

0

0

0d

0b

0

0

F16,51=67.796 p

Insecticidal and Acetylcholine Esterase Inhibition Activity of

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