The Journal of Essential Oil Research

Vol. 24, No. 3, June 2012, 279–288

Acaricidal activity of 31 essential oils extracted from plants collected in Tunisia

Sabrine Attiaa,b*, Kaouthar L. Grissab, Zeineb G. Ghrabib, Anne C. Mailleuxa, Georges Lognayc and Thierry Hancea
a
Earth and Life Institute, Biodiversity Research Centre, Université Catholique de Louvain, Louvain-la-Neuve, Belgium
b
Laboratoire d’Entomologie-acarologie. Institut National Agronomique de Tunisie, Tunis, Tunisia; cUniversité de Liège Gembloux
Agro-Bio Tech Unité de Chimie analytique, Gembloux, Belgium
(Received 25 November 2010; final form 18 October 2011)

The two-spotted spider mite Tetranychus urticae Koch is a worldwide pest, feeding on a large variety of plant fami-
lies. As its resistance to acaricides spreads rapidly, it is crucial to develop new biological control tactics to manage
its populations. In this respect, essential oils may be a good alternative, as they are currently considered minimum-
risk pesticides. In this paper, we conducted a series of laboratory experiments to determine the susceptibility of adult
females to 31 essential oils extracted from plants collected from Tunisia and compare it with the results obtained with
two synthetic acaricides (spirodiclofen and fenbutatin oxide). Details of the essential oil yield of these plants were
also recorded. A maximum yield of 0.5% was obtained. Laboratory bioassay indicated that the development of the
tetranychid mite population was significantly affected by the use of Deverra scoparia, Haplophyllum tuberculatum,
Chrysanthemum coronarium and Mentha pulegium plant extracts that killed 97%, 93%, 93% and 91% of T. urticae,
respectively. The chemical composition of the most effective extracts were characterized by gas chromatography–
mass spectrometry (GC–MS) and fast GC–flame ionization detector (FID). The evaluation of the potential of biologi-
cally active plant volatiles against T. urticae might provide a new approach to the development of natural acaricides
to be used both in biological and integrated pest management strategies for controlling two-spotted spider mites.
Keywords: Tetranychus urticae; acaricidal activity; essential oils; distillates; GC–MS; fast GC–FID

Introduction and are susceptible to temperature and UV light degra-

The two-spotted spider mite Tetranychus urticae Koch
is one of the most important pests of fruit, vegetable
and ornamental plants worldwide (1). It seriously dam-
ages outdoor crops and greenhouses (2–6). The eco-
nomic impact of this mite has increased recently
mainly because of its resistance to acaricides, which
hampers pest control (7). Indeed, synthetic acaricides
have been widely used for its control in greenhouses,
orchards and many other cropping systems, a practice
that has led to the development of resistance to acari-
cides (8–10). Consequent to this resistance, the number
of effective chemicals for the control of crop pests and
disease vectors is rapidly decreasing (11). Meanwhile,
fewer new acaricides are being introduced to the mar-
ket, largely because of the high costs associated with
research, development and registration (12). Moreover,
synthetic pesticides are known for their negative effect
on human health and the environment (13, 14).
These numerous problems are at the origin of the
recent interest in acaricidal bioactive compounds from
higher plants. Among bioactive natural compounds,
several plant essential oils are potential alternative com-
pounds for mite control (13, 15–18). Essential oils have
short residual activity and are less persistent than con-
ventional pesticides because they tend to be volatile
dation (19, 20). Consequently, most of these oils are
environmentally non-persistent and non-toxic to warm-
blooded animals and humans (21, 22) with some
exceptions, mainly due to the erroneous use of exces-
sive concentrations (23, 24). Essential oil can be
applied to field and greenhouse crops in the same man-
ner as current acaricides (25). Some can also contain
numerous secondary metabolites that deter attack from
insect and generalist herbivores (26) and provide an
alternative for resistance management as some plant
phytochemical preparations can be highly effective
against insecticide-resistant pests (27–30). Therefore,
essential oils are realistic alternatives to synthetic acari-
cides because of their selectivity, biodegradability and
few side-effects on non-target organisms and the envi-
ronment (31–36).
Many authors (17, 18, 25, 37, 38) have investigated
the use of essential oil as alternative acaricides for adult
T. urticae with promising results. Mansour et al. (39)
pointed out that, besides the toxicity, the residues of
essential oils of some Labiatae species are repelled and
strongly reduced the fecundity of Tetranychus cinna-
barinus females. Some essential oils are highly selec-
tive and specifically affect the pest populations, without
being toxic for their predators (19). In Chiasson et al.

*Corresponding author. Email: sabine_bio5@yahoo.fr

ISSN 1041-2905 print/ISSN 2163-8152 online

Ó 2012 Taylor & Francis
http://dx.doi.org/10.1080/10412905.2012.676777
http://www.tandfonline.com
280 S. Attia et al.
(38), the oil toxic effect of Chenopodium ambrosioides
was assessed on T. urticae and Panonychus citri. Other
experiments showed the acaricidal efficiency of neem
(Azadirachta indica) against Tarsonemus latus and
revealed that the population growth rate became nega-
tive when mites were exposed to plants treated with
this extract (40). Other studies pointed out that the
essential oil of Satureja hortensis L., Ocimum basili-
cum L. and Thymus vulgaris L. were effective as fumi-
gants against Bemisia tabaci and T. urticae (41).
In this study, our aim was to analyze the potential
acaricidal effects of 31 plant extracts against T. urticae
compared with that of spirodiclofen and fenbutatin
oxide, two commercial synthetic acaricides. The selec-
tion of plant species was based on the use of plant
products in traditional medicine in Tunisia (42). This
work aims to evidence the best plants for biological
control programs against the two-spotted spider mite.
Experimental
The spider mites T. urticae were collected from infected
plants from citrus orchards in Mraissa (Cap Bon Region,
north east of Tunisia) in mid-June 2006. They were
reared in the laboratory in the Institut National Agrono-
mique de Tunis (Tunisia) under controlled conditions
(temperature 25±1°C, relative humidity 60%, light:dark
16:8) and maintained for more than one year without any
contact with acaricide before experiments. The strain
was reared on bean leaves (Phaseolus vulgaris).
Selected plants and extraction technique
Thirty-one aromatic and medicinal plant species repre-
senting 17 families were collected from different Tuni-
sian localities – South (Saddine, Mednine), South-West
(Téjrouine), North-West (Kef), North-East (Hammamet,
Mraissa, Tunis) – during 2006–2008 (Table 1). Extracts
were obtained from fresh material by hydro-distillation
for 3 hours using a Clevenger-type apparatus. A few
drops of adjuvant were added (Agral 90) to extracts for
gluing the extracts (distillates or essential oil) to the
leaf surface (25). Attia et al. reported that the acaricidal
activity of these extracts was relatively enhanced with
adjuvant and exposure time (25).
Some plants extracts contained small quantities of
essential oil. In this case, we used the distillates of the
plant to assess the toxicity. Distillates were aqueous
solutions or colloidal suspensions (hydrosol) of essen-
tial oils. As essential oils, distillates were obtained by
steam distillation from plants. However, the essential
oil floated to the top of the distillate where it is
removed, leaving behind the watery distillate. In the
past, these distillates were considered a by-product of
distillation, but now are considered an important co-
product.
Essential oil yields
For each plant, we calculated the essential oil yield=oil
volume (mL) divided by the fresh weight at the begin-
ning of the tests (g) multiplied by 100 (yields are
reported in Table 2).
Part of the fresh material was weighed and then
subjected to hydrodistillation using 500 g in 3000 mL
of water. The oils were stored at 20°C until use.
Toxicity test
Toxicity tests in vitro were performed in plastic boxes,
each containing a leaf of P. vulgaris placed on water-
soaked cotton. Extracts obtained by hydro-distillation
(essential oils and distillates) were first diluted 100 times
in ethanol, then diluted again 100 times in water. After
spraying the extracts with a manual spray (spray bottle)
with about 10 mL of extract per leaf, leaves were dried
for 3 minutes. Afterwards we introduced 25 larvae in five
plastic boxes and 25 female adults in five other boxes.
Mites were included in the mortality count 72 hours
post-treatment if appendages did not move when prod-
ded with a fine pencil. We calculated the mortality
rate=(mean number of deaths after the treatment the
mean number of deaths after the control) divided by
the total number of adult female at the beginning of the
tests. For each treatment, four replications were real-
ized. Spirodiclofen and fenbutatin oxide were used as a
reference to be compared with the treatments using
plant extracts (25, 43, 44).
Identification of distillates and essential oil
constituents
The essential oils of Mentha pulegium and Chrysanthe-
mum coronarium and the distillate of Haplophyllum
tuberculatum were analyzed by gas chromatography–
mass spectrometry (GC–MS) in the Department of
Analytical Chemistry in Gembloux Agro-Biotech (Uni-
versity of Liège (Belgium)). For quantitative analyses
(percentage determination), we used a fast GC, which
proved to be powerful enough to analyze the essential
oil constituents (45, 46).
The composition of the essential oil of Deverra
scoparia has been reported in another paper by the
same authors (18).
GC–MS analysis
Conventional GC–MS analyses were carried out on a
thermo trace MS Finnigan mass-selective detector
equipped with an Optima 5 MS (Macherey-Nagel) cap-
illary column (30 m × 0.25 mm inner diameter, I.D.,
0.25 μm film thickness) and a split/splitless injector
(splitless mode) at 250°C. The oven temperature was
programmed from 40° to 210°C. Helium was the
 The Journal of Essential Oil Research 281

Table 1. List of plant species tested for their acaricidal activity, plant part used for extraction, site and date of samplings.
Plant family Scientific name Plant organ Sampled site Sampling date
Anacardiaceae Pistacia lentiscus Aerial part Saddine June 2007
Cotinus coggyra Aerial part Tunis March 2008
Apiaceae Daucus carota Aerial part Hammamet October 2006
Deverra scoparia Aerial part Téjrouine May 2008
Asteraceae Hertia cheirifolia Aerial part Saddine February 2008
Seriphidium herba-album Aerial part Saddine June 2007
Chrysantemum coronarium Flowers Tunis March 2008
Santolina Africana Aerial part Téjrouine April 2007
Cupressaceae Juniperus phoenicea Green female cones Maraissa March 2008
Fabaceae Acacia cyanophylla Flowers Tunis January 2008
Sophora secundiflora Pods and seeds Tunis March 2008
Geraniaceae Pelargonium graveolens Aerial part Hammamet March 2008
Globulariaceae Globularia alypum Leaves Saddine June 2007
Lamiaceae Salvia officinalis Aerial part Tunis March 2008
Thymbra capitata Aerial part Saddine June 2007
Rosmarinus officinalis Aerial part Saddine June 2007
Mentha pulegium Aerial part Saddine June 2007
Lavandula officinalis Aerial part Saddine June 2007
Lauraceae Laurus nobilis Leaves Hammamet January 2007
Liliaceae Allium sativum Bulbs Hammamet March 2008
Allium cepa Bulbs Hammamet March 2008
Meliaceae Melia azedarach Fruit Tunis January 2007
Myrtaceae Eucalyptus gomphocephala Leaves Saddine September 2006
Myrtus communis Aerial part Saddine June 2007
Papaveraceae Papaver rhoeas Aerial part Saddine June 2007
Lantana camara Aerial part Tunis June 2007
Rutaceae Ruta chalepensis Aerial part Saddine June 2007
Citrus aurantium Leaves Tunis March 2008
Haplophyllum tuberculatum Aerial part Mednine September 2007
Urticaceae Urtica pilulufera Aerial part Saddine October 2006
Zygophyllaceae Peganum harmala Aerial part Saddine June 2007

carrier gas at 1 mL/min. Volatile compounds were iden-
tified by comparing the mass spectra obtained with
those from the Wiley 275 L spectral library and with
their retention indices. The retention indices were deter-
mined relative to the retention times of a series of n-
alkane standards (C9–C30, Sigma-Aldrich, 0.025 μg/μL
in n-hexane), measured under the chromatographic con-
ditions described above, and compared with values in
the literature (47).
ramp 3 at 200°C/minute to 280°C, held for 0.5 minutes.
Injection temperature: 240°C; injection volume: 1 μL;
carrier gas: He, at a constant flow rate of 0.5 mL/min-
ute; split ratio=1:100. The GC unit had a high-fre-
quency fast flame ionization detector (FID; 300 Hz), at
250°C; H2 flow: 35 mL/minute; air flow: 350 mL/min-
ute; make-up gas flow (N2): 30 mL/minute. Data pro-
cessing was performed using Chromcard software
(version 2.3.3).

Fast GC analysis
Fast GC analyses were conducted on a Thermo Ultra-
Fast Trace GC gas chromatograph operated with a
split/splitless injector and a Thermo AS 3000 autosam-
pler (Thermo Electron Corp.). The GC system was
equipped with an ultra-fast module (UFM) incorporat-
ing a direct resistively heated column (Thermo Electron
Corp.): UFC-5, 5% phenyl, 5 m × 0.1 mm I.D., 0.1 μm
film thickness. The following chromatographic condi-
tions were used to obtain a suitable peak resolution.
The UFM temperature program was as follows: initial
temperature at 40°C, held for 0.1 minute, ramp 1 at 30°
C/minute to 95°C, ramp 2 at 35°C/minute to 155°C,
Data analysis
Tests were performed using Graph Pad Prism version
5.01 for windows, Graph Pad Software, San Diego,
California, USA (http://www.graphPad.com). As the
number of repetitions was four, we used non-parametric
tests (Kruskal–Wallis, Mann–Whitney). All tests were
applied under two-tailed hypotheses and the signifi-
cance level p was set at 0.05.
Results
Essential oil yields
The yield of essential oil in the current study varied
greatly. The maximum yield was obtained with D.
282 S. Attia et al.

scoparia (0.51%), Salvia officinalis (0.45%) and Cit-
rus aurantium (0.40%). With 22 plants, we obtained
enough essential oil to perform the toxicity tests. Nine
plant extracts did not yield essential oils. In this case,
we tested the toxicity of the distillates of these nine
plants (Table 2).
Mortality rate caused by the extracts
The most toxic for all stages (data of larvae and adults
grouped) were D. scoparia (98%), H. tuberculatum
(94%), fenbutatin oxide (92%), C. coronarium (91%)
and M. pulegium (91%) (Kruskal–Wallis test,
KW=6.69, p=0.15).
A more detailed analysis showed that the mortality
percentages of adults were statistically different in func-
tion of the treatments (KW=121.54, p<0.001), varying
from 1% to 97% (Figure 1). The most toxic were fen-
butatin oxide (97%), D. scoparia (97%), H. tubercula-
tum (93%), C. coronarium (93%), M. pulegium (91%),
Hertia cheirifolia (89%) and spirodiclofen (80%).
There was no difference between the efficiency of these
seven treatments (Kruskal–Wallis test, KW=11.26,
p=0.08).

Table 2. Essential oil yields according to plant species; percentages of mortality of larvae and adults in function of the plant
species.
Fresh Volume oil % % Mortality % Mortality Mann– Mann–
Plant species weight (g) (ml) Yield (larvae) (adults) Whitney (U) Whitney (P)
Acacia cyanophylla D 12000 0.5 0 58 26 0.0 0.0⁄
Allium cepa E 9000 1.2 0 65 67 7.0 0.8
Allium sativum E 9000 1 0 86 61 0.0 0.0⁄
Seriphidium herba- 3614 10 0.2 54 37 0.5 0.0⁄
album E
Bolido 78 80 7.0 0.8
Chrysanthemum 7300 5 0 88 93 4.5 0.3
coronarium E
Citrus aurantium E 1500 6 0.4 63 55 4.5 0.3
Cotinus coggyra D 3000 0 0 58 58 7.5 1.0
Juniperus phoenicea E 3500 2.5 0 60 56 4.0 0.3
Daucus carota D 423 0 0 5 3 6.5 0.7
Eucalyptus 5300 2 0 60 34 0.0 0.0⁄
ghomphocephala E
Pelargonium 450 1.5 0.3 78 70 3.5 0.2
graveolens E
Globularia alypum D 1500 0 0 8 2 4.0 0.2
Haplophyllum 2000 2.5 0.1 94 93 7.5 1.0
tuberculatum D
Hertia chaerifolia E 1705 2 0.1 81 89 3.5 0.2
Lantana camara D 1732 0 0 2 1 6.0 0.6
Laurus nobilis E 800 2 0.2 63 46
Lavandula officinalis E 1200 2.5 0.2 38 41 0.5 0.0⁄
Melia azedarach D 526 0 0 77 75 6.5 0.7
Mentha pulegium E 3325 6 0.1 90 91 7.0 0.8
Myrtus communis E 6200 3 0 82 47 7.0 0.8
Santolina africana E 500 1 0 77 68 0.0 0.0⁄
Papaver rhoeas D 359 0 0 43 34 2.5 0.1
Peganum harmala D 1500 0 0 34 12 2.0 0.1
Pistacia lentiscus E 1500 1 0 22 23 0.0 0.0⁄
Deverra scoparia E 1166 6 0.5 98 97 7.5 1.0
Rosmarinus officinalis 4203 15 0.3 61 53 7.0 0.8
E
Ruta chalepensis E 3000 0.5 0 66 61 4.5 0.3
Salvia officinalis E 1250 5.5 0.4 61 57 3.5 0.2
Sophora secundiflora 8500 0 0 68 61 6.0 0.6
D
Control 1 1 3.5 0.2
Thymbra capitata E 7339 35 0.4 61 52 8.0 0.8
Torque S 87 97 4.0 0.3
Urtica pilulifera D 700 0 0 49 46 2.0 0.1

Notes: These percentages were compared with the results obtained in the control test; the statistical tests were Mann–Whitney, with the U and P
values in the two last columns. D, distillate; E, essential oil.
 The Journal of Essential Oil Research 283

The mortality percentages of larvae were also statisti- (41.86%), followed by menthone (28.33%) and 3-octa-
cally different in function of the treatment (KW=127.6, nol (6.93%) (Table 4).
p<0.001), varying from 1% to 98% (Figure 2). The most Moreover, GC–MS analysis indicated that in the C.
toxic were D. scoparia (98%), H. tuberculatum (94%), coronarium oil, nine constituents comprised 96.38% of
M. pulegium (90%), C. coronarium (88%), fenbutatin the oil by weight. Pulegone was the most abundant
oxide (87%), Allium sativum (86%), Myrtus communis compound (29.76%), followed by camphor (27.55%)
(82%) and H. cheirifolia (81%). There was no difference and isomenthone (17.65%) (Table 5).
between the efficiency of these eight treatments (Krus-
kal–Wallis test, KW=12.17, p=0.09). Discussion
Most treatments were similarly efficient on larvae
and adults with the noteworthy exception of seven plants In this study, we assess the toxicity of 31 plant
extracts (Acacia cyanophylla, A. sativum, Seriphidium extracts (distillates and essential oils) collected in
herba-album, Daucus carota, H. cheirifolia, Melia azed- Tunisia and of two synthetic acaricides (Spirodiclofen
arach and Santolina africana) that were more toxic on and fenbutatin oxide) against T. urticae larvae and
larvae than on adults (Table 2). Concerning the synthetic adults. Some plants are endemic to the North African
acaricide, the efficiency of fenbutatin oxide was always Sahara, such as D. scoparia, S. africana and H. cheiri-
comparable with the most toxic plant extracts. Spirodi- folia (48–50); some can be found throughout the Med-
clofen was efficient against the adults (80%) but, curi- iterranean circumference such as Thymbra capitata,
ously, was not that efficient against larvae (78%). Rosmarinus officinalis, S. herba-album and Pelargo-
nium graveolens (48). Some of these plants were
never tested previously as a biological control against
Components analysis of extracts: T. urticae; this is the first study demonstrating that H.
Tables 3-5 show results from GC–MS and fast GC– tuberculatum, H. cheirifolia, C. aurantium and S. afri-
FID analysis for H. tuberculatum, M. pulegium and C. cana have acaricidal activity.
coronarium, the most active extracts against T. urticae. The yield of essential oil varied considerably
Through comparison of the retention indices with depending upon plant species. Here we show that the
those of the literature (47) and electron impact (EI) plants that contain the most interesting quantity of oil
mass spectra of each peak with the library, it was possi- are D. scoparia (0.51%), S. officinalis (0.45%) and C.
ble to identify 20 individual components of Haplophyl- aurantium (0.40%). Maximum yields of 0.5% were
lum distillate comprising 99.54% of the total weight. achieved with the essential oil of D. scoparia. Nine
1-Terpineol was the most abundant compound plant extracts do not contain essential oils at all. We
(23.82%), followed by piperitone (15.03%) and carva- think that this low yield is not a limit to a large-scale
crol (12.68%) (Table 3). application possibility of some plant extracts and that it
Similarly, there were eleven known constituents in is possible to use distillates to control T. urticae. As we
the M. pulegium oil, comprising 99.95% of the total show in this paper, H. tuberculatum distillate is toxic
weight. Pulegone was the most abundant compound for 94% and 93% of adult and larvae T. urticae,

* ** * ns

% mortality

100
90
80
70
60
50
40
30
20
10 Plant extracts
0
Fenbutatin oxide
D. scoparia
C. coronarium
H. tuberculatum
M. pulegium
H.cheirifolia
Spirodiclofen
M.azedarach
P.graveolens
S. africana
A. cepa
S.secundiflora
A. sativum
R. chalepensis
C.coggyra
S. officinalis
J. phoenicea
C. aurantium
R. officinalis
T. capitata
M. communis
U. pilulifera
L. nobilis
L. officinalis
S. herba-album
E.ghomphocephala
P. rhoeas
A. cyanophylla
P.lentiscus
P.harmala
D.carota
G.alypum
L.camara
Control

Figure 1. Ranking of the mean percentage of mortality Tetranychus urticae adults after 72 hours exposure to extracts. Mean
percentages are significantly different at ⁄p60.05, ⁄⁄p60.01 and ⁄⁄⁄p60.001; ns, not significant; Kruskal–Wallis test.
284 S. Attia et al.

*** ** * ns

% mortality
100
90
80
70
60
50
40
30
20
10
0 Plant extracts
D. scoparia

H.tuberculatum

M.pulegium

C.coronarium

Fenbutatin oxide

A.sativum

M.communis

H.cheirifolia

P.graveolens

Spirodiclofen

S.africana

M.azedarach

S.secundiflora

R. chalepensis

A.cepa

L.nobilis

C.aurantium

T.capitata

S.officinalis

R.officinalis

J. phoenicea

E.ghomphocephala

C.coggyra

A.cyanophylla

S. herba-album

U.pilulifera

P. rhoeas

L. officinalis

P.harmala

P.lentiscus

G.alypum

D.carota

L.camara

Control
Figure 2. Ranking of the mean percentage of mortality Tetranychus urticae larvae after 72 hours exposure to extracts. Mean
percentages are significantly different at ⁄p60.05, ⁄⁄p60.01 and ⁄⁄⁄p60.001; ns, not significant; Kruskal–Wallis test.

Table 3. Major constituents of Haplophyllum tuberculatum and their relative proportions in the pure distillate identified by gas
chromatography–mass spectrometry (GC–MS; %: compound percentage) and quantified by fast GC–flame ionization detector
(GC–FID).
No. Components Retention index %
1 Sabinene 975 0.7
2 2-Octanol 995 0.9
3 α-Phellandrene 1003 2
4 trans-Linalool oxide (furanoid) 1073 5.5
5 cis-Linalool oxide (furanoid) 1087 0.2
6 Terpinolene 1089 6.8
7 Linalool 1097 0.3
8 1-Terpineol 1134 23.8
9 Camphor 1146 5.2
10 Borneol 1169 3.3
11 Terpinen-4-ol 1177 2.7
12 para-Cymen-8-ol 1183 0.2
13 α-Terpineol 1189 2
14 Citronellol 1226 8.9
15 Pulegone 1237 0.3
16 Geraniol 1253 7.8
17 Piperitone 1253 15
18 Thymol 1290 0.2
19 Carvacrol 1299 12.6
20 Eugenol 1359 0.4

respectively. Distillates are widely available and are toxic. Among these plants extracts, the best candidate
especially interesting, as they are not very expensive in for an efficient control seems to be D. scoparia. This
comparison with essential oils. Moreover, their extrac- plant is endemic to North Africa and is particularly
tion is very easy and does not require a high quantity active against the larvae and adults of T. urticae, caus-
of plants, contrary to essential oils. Further develop- ing 97% mortality in adults and 98% mortality in lar-
ment will be needed to implement the applications of vae. Moreover, D. scoparia extracts contain enough
plants extracts in orchards, greenhouses and field essential oil to perform our tests. As shown in a previ-
(essential oils and/or distillates) and to look toward ous paper (18, 25), the essential oil of D. scoparia is
more efficient methods of extraction and plant culture. toxic to T. urticae with LC90 and LC100 values of 3.2
Our results show that D. scoparia, H. tuberculatum, and 3.42 mg/L (18). The exact active site of this essen-
C. coronarium and M. pulegium have great potential for tial oil on the metabolism of two-spotted spider mites
effective management of T. urticae, as they are the most is still unknown.
 The Journal of Essential Oil Research 285

Table 4. Major constituents of Mentha pulegium and their relative proportions in the pure oil identified by gas chromatography–
mass spectrometry (GC–MS;%: compound percentage) and quantified by fast GC–flame ionization detector (GC–FID).
No. Components Retention index %
1 α-Pinene 939 0.5
2 β-Pinene 979 1
3 3- Octanol 991 6.9
4 Limonene 1029 9
5 Menthone 1153 28.3
6 Pulegone 1237 41.8
7 Piperitone 1253 4.9
8 Carvacrol 1299 3.3
9 Piperitenone 1343 1.7
10 α-Humulene 1455 1.5
11 Humulene epoxide II 1608 0.6

Table 5. Major constituents of Chrysanthemum coronarium and their relative proportions in the pure oil identified by gas
chromatography–mass spectrometry (GC–MS;%: compound percentage) and quantified by fast GC–flame ionization detector (GC–
FID).
No. Components Retention index (literature) %
1 Camphene 954 2.6
2 Δ-3-Carene 1031 2.3
3 Alloocimene 1132 2.3
4 Camphor 1146 27.5
5 Menthone 1153 3.0
6 Isomenthone 1163 17.6
7 Pulegone 1237 29.7
8 cis-Chrysanthenyl acetate 1265 7.1
9 Bornyl acetate 1289 3.9

The toxicity effect of D. scoparia may also be
explained by higher -pinene and sabinene contents (18,
51). Ten major components, comprising 98.52% of the
total weight were identified; -pinene was the most
abundant constituent (31.95%) followed by sabinene
(17.24%) and Δ-3-carene (16.85%) (18). The report of
Attia et al. strongly suggested that D. scoparia oil
might have more than one mode of action (fecundity
and mortality) (18).
α-Pinene, the major constituent (31.95%) of D.
scoparia oil, was the major contributor to the toxicity
of the artificial blend of the essential oil, followed by
Δ-3-carene (16.85%) and terpinene-4-ol (2.84%).
Indeed, three major constituents (myrcene, eugenol and
Δ-3-carene) strongly decreased fecundity (18).
In the literature, there are many studies on the
chemical composition and various activities of the
essential oil of Haplophyllum robostum (52) and
Haplophyllum ramosissimum (53), but few studies were
done to analyze the composition of the essential oil of
H. tuberculatum (52,54). In this study, the chemical
constituents of the essential oil of the aerial part of H.
tuberculatum were studied. The major components in
H. tuberculatum distillate were identified as 1- terpineol
(23.82%), piperitone (15.03%) and carvacrol (12.68%)
(Table 1). The distillate composition herein evaluated
was different to the natural oil characterized by Al-
Burtamani et al. (52). H. tuberculatum distillate is
mainly composed of monoterpenes hydrocarbons with
formula (C5H8)2 (Table 1). The total C10 dienes in
Haplophyllum oil was about 46.5% (52).
Concerning C. coronarium and M. pulegium, the
two oils differ in their most abundant compounds. The
most abundant compounds of the Chrysanthemum oil
were pulegone (29.76%), camphor (27.55%) and isom-
enthone (17.65%); pulegone (41.86%), menthone
(28.33%) and 3-octanol (6.93%) were characteristic of
the Mentha oil (Tables 4 and 5).
The formation of essential oil in the plant, and con-
sequently the yield and composition of the oil pro-
duced, depends on many factors. Genetic differences in
plants of the same species that are otherwise indistin-
guishable (chemotypes) can result in widely different
essential oil content.
According to Miresmailli et al. (55), the major
chemical components of essential oils with acaricidal
activity have been identified as 1,8 cineole, α-pinene,
and myrcene. The differences of toxicity between plant
species are probably related to differences in chemical
composition and also of proportion of main compo-
286 S. Attia et al.
nents present in extracts (17, 18, 34, 56, 57). The next
step of this study should be to study the toxicity of
each constituent of the most effective essential oils
separately.
Plant extracts can have lethal effects and can also
affect the fecundity of females (17, 18, 39). They might
also present other interesting properties from a biologi-
cal control point of view. For instance, some plant
extracts affect the behavior of the pests as they are
repellent for insects or acari (38, 39, 58).
For instance, garlic extracts are repellent to T. urti-
cae. Some extracts can have a very selective effect and
specifically affect the pest populations, without being
toxic for their predators (19). Furthermore, pure rose-
mary oil and Eco/Trol (rosemary oil-based pesticide)
caused complete mortality of spider mites T. urticae at
concentrations that are not phytotoxic to the host plant
and that did not cause any mortality in T. urticae preda-
tors such as Phytoseilus persimilis (19). On the con-
trary, some can also be more general biocides and
affect animals from different orders. For instance,
Anthony et al. (59) showed the biocidal properties of
the essential oils of H. tuberculatum and Plectranthus
cylindraceus against Meloidogyne javanica (Nematoda).
These side-effects can be interesting from a biological
point of view, but they can also dramatically affect the
equilibrium between different species. Therefore, the
other potential properties of plants toxic for pests must
be studied before being used in an integrated program
of pest control. Another step of this work should be to
study the plant extracts advantages and drawbacks from
an economical point of view and to compare them with
chemical applications effects, taking account of envi-
ronmental pollution and the potential apparition of
resistance in pest population (60).
Conclusion
Among 31 plants tested as acaricides against the phy-
tophagous mite T. urticae (Boisduval) in laboratory tri-
als, the most toxic were D. scoparia, H. tuberculatum,
M. pulegium and C. coronarium. One of these highly
effective plants is endemic to North Africa: D. scoparia.
The present research proved that D. scoparia, H.
tuberculatum, H. cheirifolia, C. aurantium and S. afri-
cana have acaricidal activity against T. urticae.
Future research efforts should be directed towards
the improvement of processing and extraction methods
to obtain extracts of high grade, preserving and/or
improving its peculiar biological properties.
Acknowledgements
We are very grateful to George van Impe and Le goff
Guillaume for the useful discussions about T. urticae. The
authors are also indebted to the Wallonie-Bruxelles
International (WBI). This is publication BRC 192 of the
Biodiversity Research Centre at UCL.
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