Veterinary Parasitology 214 (2015) 348–352

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Veterinary Parasitology
journal homepage: www.elsevier.com/locate/vetpar

Short communication

The repellent and persistent toxic effects of essential oils against the
poultry red mite, Dermanyssus gallinae
I.S. Nechita a,b , M.T. Poirel b,c , V. Cozma a , L. Zenner b,c,∗
a
University of Agricultural Sciences and Veterinary Medicine, Department of Parasitology, Avenue Manastur 3-5, 400372, Cluj-Napoca, Romania
b
VetAgro Sup, Veterinary Campus of Lyon, University of Lyon, Department of Parasitology, 1 avenue Bourgelat, F-69280 Marcy-Etoile, France
c
Laboratory of Biometrics and Evolutionary Biology, CNRS UMR 5558, University of Lyon, 43 Boulevard of 11 November, 69622 Villeurbanne, France

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

Article history: The economic impact of the poultry red mite, Dermanyssus gallinae, the lack of new acaricides, the occur-
Received 31 March 2015 rence of resistance and tighter legislation have all led to the need to find new ways to control this pest. One
Received in revised form 7 October 2015 promising alternative method of control focuses on employing repellent and/or toxic effects of selected
Accepted 14 October 2015
plant essential oils against D. gallinae. Ten essential oils (basil, thyme, coriander, eucalyptus, lavender,
lemon, fir tree, oregano, mint, and juniper) were tested for the persistence of toxic and repellent effects.
Keywords:
In filter-paper toxicity bioassays against D. gallinae, the best results were observed for lavender (more
Dermanyssus gallinae
than 97% mortality after 48 and 72 h) and thyme (84% at 72 h) at a dose of 0.12 mg/cm2 . In addition, two
Essential oil
Toxicity
oils showed significant persistent toxic effects 15 and 30 days post application to filter papers. Thyme
Repellent was the most effective (100% mortality at 72 h), followed by lavender (nearly 80% mortality after 72 h).
Persistent Out of the ten oils tested for their repellent effect, thyme was the strongest, with nearly 80% of the tested
area avoided by mites; oregano caused a 60% avoidance and lavender exhibited an effect close to 40%.
All other oils exhibited a repellent effect of less than 30%. None of the experiments showed a repellent
effect for HM (commercial alimentary oil) or negative controls. We found that the thyme and lavender
essential oils exhibited promising results when tested in vitro for toxic and repellent effects against D.
gallinae; thus, we suggest that future experiments focus on in vivo tests using these oils in farm units.
© 2015 Elsevier B.V. All rights reserved.

1. Introduction
Dermanyssus gallinae (De Geer, 1778), known as the “poul-
try red mite,” is considered one of the most important welfare
and health problems for poultry, particularly laying hens (Chauve,
1998; Emous et al., 2005; Hegelund and Sorensen, 2007) and
causes large economic losses worldwide (Bruneau et al., 2001). This
hematophagous mite parasitizes chickens, residing on its host only
during blood meals, then retreating into hiding in farm buildings. A
characteristic of red mite infestation is that once they are present in
a flock, it is almost impossible to eradicate them (Chauve, 1998). The
control of the poultry red mite is mainly based on the use of aca-
ricides, such as organophosphates, carbamates, pyrethroids, and
formamidines. The lack of new acaricides, resistance to acaricides,
and increasingly stringent requirements for chemicals used on food
∗ Corresponding author at: VetAgro Sup, Veterinary Campus of Lyon, University
of Lyon, Department of Parasitology, 1 avenue Bourgelat, F-69280 Marcy-Etoile,
France.
E-mail address: lionel.zenner@vetagro-sup.fr (I.S. Nechita).
animals greatly limit the options for controlling this pest (Zeman,
1987; Ash and Oliver, 1989; Beugnet et al., 1997; Sparagano et al.,
2009). Therefore, alternative methods for the control of D. gallinae
are urgently needed (Birkett et al., 2008).
One alternative method focuses on employing the repellent
and/or toxic effects of selected plant-derived products (PDPs), par-
ticularly essential oils, for the management of numerous pests,
including D. gallinae. In vitro tests have shown promising activities
of PDPs against many food-borne pathogens and spoilage microor-
ganisms. Some studies have investigated the toxicity of essential
oils against different livestock pests, such as the study by Kim et al.
(2004, 2007), which tested 56 plant essential oils for their acaricidal
effects against D. gallinae. More recently, George et al. (2008, 2009,
2010) investigated the influence of ‘time since last blood meal’ on
the toxicity of essential oils to the poultry red mite, the repellence
of plant essential oils to D. gallinae, and their toxicity to the non-
targeted invertebrate Tenebrio molitor, as well as the in vitro and
in vivo acaricidal activity and residual toxicity of spinosad against
the poultry red mite.
To develop novel and reliable alternative methods for the man-
agement of this pest, we investigated ten essential oils with regard

http://dx.doi.org/10.1016/j.vetpar.2015.10.014
0304-4017/© 2015 Elsevier B.V. All rights reserved.
 I.S. Nechita et al. / Veterinary Parasitology 214 (2015) 348–352
to their acute toxicity at 24–72 h, the persistence of their toxicity
after 15 and 30 days, and their repellent effects against D. gallinae.
For this study, we selected ten plant essential oils (basil, thyme,
coriander, eucalyptus, lavender, lemon, fir tree, oregano, mint, and
juniper) that are frequently used in the management and control
of numerous pests.
2. Materials and methods
2.1. Mites
D. gallinae mites were collected from a laying hen facility in the
Lyon region. In the laboratory, the mites were transferred into an
open jar surrounded by two successive barriers of water and were
then subsequently left for 10 to 15 days before the start of exper-
iments to stabilise the population and to acclimatise the parasites
to the laboratory conditions.
2.2. Selection of plant essential oils
All plant essential oils used in this study were produced by
SOLARIS PLANT SRL (Bucharest, Romania) and were purchased from
a herbal pharmacy (Cluj-Napoca).
2.3. Bioassays
Filter-paper contact bioassays were used to evaluate the toxicity
and persistence of the toxic and repellent effects of the essential oils
towards D. gallinae.
For the toxicity bioassay, a 20% dilution of each essential oil in
distilled water was made with 0.12 mg/cm2 of the oil. Control filter
papers (Whatman No. 2) contained nothing, commercial alimen-
tary oil (Huile de mélange Omegabio 4 (HM), Cauvin, France) as a
negative control, or Phoxim at a dose of 0.52 mg/cm2 (ByeMite nd,
Bayer) as a positive control. After drying in a fume hood, each filter
paper (length: 8 cm; width: 1 cm) was folded length-wise and was
placed into a Pasteur pipette, allowing mites to walk on the impreg-
nated paper or on the glass wall of the pipettes. Batches of 18–25
adult D. gallinae were introduced into the pipettes using a vacuum
pump. The fine ends of the pipette were broken and covered with
parafilm, and the other ends were plugged with cotton. For each
experiment, three replicates were performed for each essential oil
in addition to two negative controls (paper without anything or
HM) and a positive control (Phoxim, ByeMite). All pipettes were
checked to ensure that no dead mites were introduced and to verify
the number of mites in each pipette. All pipettes were maintained
under the same conditions as those used for batch maintenance.
The mites were then examined after 24, 48, and 72 h of exposure
to the essential oils to determine mite mortality using a binocu-
lar microscope. All experiments were repeated twice. The results
were expressed as the percentage mortality and the means and
standard deviations for each of the oils or controls were calculated,
and the means were then corrected using Henderson–Tilton’s for-
mula (Henderson and Tilton, 1955), which relies on a non-uniform
mite population. The persistence was tested after 15 and 30 days
using the same procedure as described above with filter papers
introduced into Pasteur pipettes and left under a hood for 15 or 30
days before the experiments.
To evaluate the repellent effect of essential oils toward D. gal-
linae, we used a video camera to register the movement of mites
exposed to essential oils for 30 min. In this experiment, a circular
rubber ring (4.5 cm diameter) was placed onto a filter paper cut to
the same size, and both were placed between two pieces of glass.
On each filter paper, two circles (0.81 cm diameter) were previ-
ously drawn at two opposite points of the ring, 2 ␮L of each tested
essential oil or control (nothing or HM) was placed in the centre
349
of one of these circles, and 2 ␮L HM oil was placed in the centre
of the remaining circle as a control. Prior to mounting the appara-
tus, the filter papers were dried under a fume hood, and a single
mite was then inserted just before the experiment. The video cam-
era recorded 30 min of each test. After each test, the pieces of glass
and rubber circle were washed with detergent and alcohol prior to
reuse. For this experiment, all the essential oils were tested against
HM at 100% concentration, or nothing, and three oils (thyme, laven-
der, and oregano) were tested at different concentrations (2, 5, and
20%). For each experiment, the trajectory of the mite in the appara-
tus was recorded on paper, and the area avoided by the mite during
the experiment was measured. Resting periods were also noted, to
exclude mites that rested for more than half the duration of the
experiment (15 min) from the results. By relying on the measured
area and surface of the ring, we calculated the RE (repellent effect)
as a percentage of the area avoided by the mite for each experiment.
Means and standard deviations for each of the oils or control were
then calculated. For each of the oils or control, five replicates were
performed.
3. Results
3.1. Toxicity and persistence of essential oils
The toxic effect of 20% dilutions of ten essential oils at an appli-
cation rate of 0.12 mg/cm2 , varied according to the plant species.
At this dose, the best results were observed for lavender (more
than 97% mortality after 48 and 72 h) and thyme (84% at 72 h). The
oregano and juniper oils resulted in a mortality rate close to 50%
after 72 h, whereas the other oils resulted in a lower or no mortality
compared with the controls (Fig. 1A).
In addition, the persistence of the toxic effect of these oils after
15 and 30 days from application on filter paper varied according
to the plant species. As shown in Fig. 1, only three oils (lavender,
oregano, and thyme) showed significant activity 30 days after appli-
cation to filter papers, with lavender being less effective (nearly 80%
mortality after 72 h), thyme the most effective (100% at 72 h), and
oregano the least effective, with more than 40% mite mortality after
72 h (Fig. 1C).
3.2. Toxicity of thyme and lavender at different concentrations
Based on the results obtained at the 20% concentration level
for essential oils, we tested the thyme and lavender oils at differ-
ent concentrations (2, 5, and 10%) to establish whether these oils
maintained their acaricidal activity. Thyme oil exhibited a greater
activity at 20% (100% at 48 and 72 h) compared to lavender oil (80%
at 72 h), and at 5% dilution, more than 60% activity at 48 and 72 h
was observed for thyme and only >40% activity at 72 h for laven-
der. At 10% dilution, lavender was more effective (>80% at 48 and
72 h) than thyme (>60%). For the final dilution (2%), both oils had
approximately the same effect.
3.3. Repellent effect of essential oils
All ten essential oils were tested for their repellent effect by
using a drop of 2 ␮L of pure oil. Out of the ten oils, thyme had the
strongest repellent effect, with nearly 80% of the area avoided by the
mites; oregano resulted in a 60% avoidance and lavender approxi-
mately a 40% avoidance. All of the other oils exhibited a repellent
effect of less than 30%. None of the experiments showed a repellent
effect for the HM or negative controls (Fig. 2A and 2B).
350 I.S. Nechita et al. / Veterinary Parasitology 214 (2015) 348–352

A. 100

80

60
h24
40
h48
20
h72
0

B. 100

80

60
h24
40
h48
20
h72
0

C. 100

80

60
h24
40
h48
20 h72
0

Fig. 1. The acaricidal activity of 10 essential oils against Dermanyssus gallinae using the filter-paper contact bioassay at 0.12 mg/cm2 of the oil.
A—Corrected means and standard deviations of the toxic effect at D0 (day 0).
B—Corrected means and standard deviations of the toxic effect at D15 (after 15 days).
C—Corrected means and standard deviations of the toxic effect at D30 (after 30 days).

3.4. Repellent effect of thyme, lavender, and oregano at different
concentrations
Based on the results obtained from the toxicity experiments,
we chose thyme, lavender, and oregano to examine their repellent
effects at different concentrations (100%, 20%, 5%, and 2%). Similar
to previous results, thyme was the most repellent at the 100% con-
centration, followed by oregano and lavender. This order was also
maintained for the 20% concentration, but the effects rapidly and
constantly decreased, with little or no effect at 2% for the three oils,
with the exception of lavender, which still had a slight effect at 2%
(Fig. 2A and C).
4. Discussion
One of the alternative methods for the control of different
culture breeding pests is the use of plant essential oils, which
have been recognised for their properties (antibacterial, antifun-
gal, antiviral, insecticidal, and antioxidant) and are widely used in
medicine and the food industry.
In comparison to other studies, we made four dilutions (2%, 5%,
10%, and 20%) of the plant essential oils in distilled water and placed
them in filter paper in Pasteur pipettes. Lavender and thyme were
most effective at a dose of approximately 0.12 mg/cm2 of the oil,
corresponding to a 20% dilution and >80% mortality. In previous
studies (George et al., 2009; Kim et al., 2004), the same principle
and method was used to establish the toxicity of essential oils, and
the only difference was that Petri dishes were used in one study
and glass vials in the other, and adult mites were then introduced
using flasks.
A problem with the use of essential oils against different pests is
that oils can have different chemical profiles and therefore, varying
acaricidal activity, even if the oils are from the same plant species.
Kim et al. (2004) found that lime, thyme, lavender, oregano, and
coriander led to 100% mite mortality in contact toxicity assays at
0.07 mg/cm2 of the oil. George et al. (2009) reported that these oils
were less effective than in a previous study, and in our study, these
oils had moderate toxicity, with only lavender resulting in >90%
mite mortality. In addition, George et al. (2010) found that not all
essential oils have the same toxic effect on all of the life stages
 I.S. Nechita et al. / Veterinary Parasitology 214 (2015) 348–352 351

Fig. 2. The repellent effect of 10 essential oils against Dermanyssus gallinae.

A—Sketches showing the movement of mites in the presence of the essential oils and the avoided area.
B—The means and standard deviations of the area avoided by mites, due to the repellent effect of essential oils.
C—The means and standard deviations of the area avoided by mites, due to the repellent effect of thyme, lavender and oregano oils at different dilutions: 100%, 20%, 5% and
2%.

of D. gallinae, and thyme oil in the previous study produced 100%
mortality in adults but had no effect on eggs. Kim et al. (2004) found
that not all of the oils from the same species had the same effect,
as in the case of Eucalyptus spp., juniperspp., mint spp., oreganospp.,
and seven Citrusspp, of which lime oil produced 100% mortality and
the others were almost ineffective at the same dose. George et al.
(2008) reported that the acaricidal effect of Lavendula angustifolia
from Croatia was nearly 1.5 times more lethal to D. gallinae than
the same oil from the same species from France.
All of the plant essential oils used in the present study were
found to have different levels of toxicity forwards D. gallinae, and
differences in the repellent effect between the oils used in this study
were observed. For example, lavender oil resulted in the highest
mite mortality at >90%, and the mortality caused by thyme oil was
>80% when tested for toxicity, although thyme oil had the strongest
repellent effect, followed by oregano, lavender, and basil oil.
Regarding the repellent effect of essential oils against D. galli-
nae, only one study exists (George et al., 2009), with which we can
compare our results. To test the repellent effect of essential oils,
these authors used four Y-tube olfactometers of the same design,
which were assembled using flexible plastic tubing coupled to hol-
low, rigid glass rods and plastic bungs. In this study, we used a
video camera to record the movement (trajectory) and periods of
rest of the mite for a 30-min period, between two pieces of glass.
We obtained repellent effect results faster using this method, and
we used only one mite; thus, our results might be more accurate
than those of others that used a group of mites, but might require
more replication. However, using this method, we could not deter-
mine precisely whether the repellent effect of essential oils occurs
352 I.S. Nechita et al. / Veterinary Parasitology 214 (2015) 348–352
via direct contact or in the vapour phase; for the Y-tube olfactome-
ters that were used by George et al. (2009), the repellent effect was
manifested in the vapour phase.
George et al. (2009) found that thyme oil was the most repellent
of all oils tested and maintained its repellent effect for more than 13
days, and that thyme oil had the most highly toxic effect. Thymol,
the main constituent of thyme oil, has a repellent effect against
Psoroptes cuniculi (Perruci et al., 1995), suggesting that thyme oil
could be used not only on D. gallinae but also on other mites species.
In our experiments, thyme essential oil also had the strongest repel-
lent effect according to the area avoided by the mites, which was
more than 80%, followed by oregano oil (60%) and lavender oil
(nearly 40%). We also tested the repellent effect at different dilu-
tions (20%, 5%, and 2%) and found that it decreased concomitantly
with an increasing dilution.
This study showed that thyme and lavender essential oils exhib-
ited promising results when tested in vitro for toxic and repellent
effects against D. gallinae, and further experiments should focus
on in vivo tests of these oils in farm units to confirm these results.
In vitro trials that combine different essential oils should be used
to determine whether their effects become stronger than those
observed here, or decrease. In addition, these protocols could lead
to in-depth studies of the behaviour of this pest and help to identify
novel ways to control it.
Acknowledgements
The authors gratefully thank the Department of Parasitology,
VetAgro Sup, Université de Lyon for their support. This study was
supported by the Erasmus Mundus Programme, University Joseph
Fourier, VetAgro Sup, University of Agricultural Sciences and Vet-
erinary Medicine, CNRS, and the University of Lyon.
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