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Journal of Vector Ecology June 2009

Efficacy of the botanical repellents geraniol, linalool, and citronella against


mosquitoes
Günter C. Müller1, Amy Junnila2, Jerry Butler3, Vassiliy D. Kravchenko4, Edita E. Revay5,
Robert W. Weiss6, and Yosef Schlein1
1
Department of Parasitology, Kuvin Centre for the Study of Infectious and Tropical Diseases, The Hebrew University,
Hadassah-Medical School, Jerusalem, Israel, 91120
2
Department of Parasitology, McGill University, Macdonald Campus, Ste-Anne-de-Bellevue, Quebec H9X 3V9, Canada
3
Medical-Veterinary Entomology and Nematology Department, Institute of Food and Agricultural Sciences,
University of Florida, Gainesville, FL 32611-0620, U.S.A.
4
Department of Zoology, Tel Aviv University, Tel Aviv, Israel
5
Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel
6
Entomological Associates, 102 Beaver Mills Road, Julian, PA 16844, U.S.A.

Received 25 February 2007; Accepted 2 December 2008

ABSTRACT: We determined the degree of personal protection provided by citronella, linalool, and geraniol in the form of
commercially available candles or diffusers, both indoors and outdoors. Under the uniform conditions of the experiments,
all substances repelled significantly more mosquitoes than the unprotected control. Furthermore, the repellents tested were
more active when in the form of a continuous release diffuser than in candle form. All candles were 88g containing 5% of the
active ingredient and all diffusers contained 20g of 100% active ingredient. Indoors, the repellency rate of citronella candles
was only 14% while the repellency rate of citronella diffusers was 68%. The repellency of geraniol candles was 50% while
the diffusers provided a repellency rate of 97%. No linalool candles were available for study but linalool diffusers repelled
mosquitoes by 93%. Outdoors, citronella diffusers placed 6 m from mosquito traps repelled female mosquitoes by 22%,
linalool repelled females by 58%, and geraniol repelled females by 75%. Trap catches were significantly reduced again when
diffusers were placed 3 m from the traps. We concluded that geraniol had significantly more repellent activity than citronella
or linalool in both indoor and outdoor settings. Journal of Vector Ecology 34 (1): 2-8. 2009.

Keyword Index: Monoterpene alcohols, mosquito repellent, citronella, geraniol, linalool, Aedes spp., Culex spp., botanical
repellents, plant essential oils, Puerto Rico.

an advisory committee in 2000 to curb inconsistencies in


INTRODUCTION repellent product performance testing. The committee
pointed out that many factors play a role in how effective
In recent years, botanical insect repellents have become repellents are, including frequency and uniformity of
increasingly popular as viable alternatives to synthetic repellent application (Khan et al. 1972, Gabel et al.1976),
chemical pest repellents because they reputedly pose little the number and species of the organisms attempting to bite,
risk to the environment or human health. However, the the user’s inherent attractiveness to blood-feeding insects,
body of scientific literature documenting bioactivity of age (Muirhead-Thomson 1951), sex (Gilbert et al. 1966)
plant derivatives is sometimes contradicting and lacking in and size of the potential host (Port and Boreham 1980), and
standardized testing protocols. Therefore, we feel there is a the physical activity level of the potential host. As well, the
need for increased research on the use of natural or herbal- committee made recommendations as to the duration of
based repellents to ensure quality and determine the most the tests, statistical significance of sample size, replication
effective means of application and use. of experiments, and rotation of human volunteer subjects
N,N-diethyl-3-methylbenzamide (DEET) remains (http://www.epa.gov/oscpmont/sap/meetings/2000/april/
the gold standard of currently available insect repellents, freportapril572000.pdf).
however, there have been case reports of DEET toxicity in Specifically, many claims have been made regarding the
the literature (Zadikoff 1979, Snyder et al. 1986, Osimitz and repellent properties of citronella essential oil and various
Grothaus 1995, Osimitz and Murphy 1997) and consumer terpene alcohols (Hwang et al. 1985, Tawatsin et al. 2001,
interest in natural alternative repellents is growing rapidly. Barnard and Xue 2004). Citronella essential oil is derived
A broad spectrum of plants and plant essential oils have from different species of Cymbopogon (citronella grass) and
been tested as potential insect repellents (for a review see contains some industrially important aromatic compounds
Sukumar et al. 1991). Out of those tested, such a large number such as geraniol. Geraniol and linalool, isomers of each other,
of claims of repellent activity were made that the United are monoterpene alcohols found in many plant essential oils
States Environmental Protection Agency (EPA) formed such as citronella and thyme respectively (Choi et al. 2002,
Vol. 34, no. 1 Journal of Vector Ecology 3

Park et al. 2005). Unfortunately, there seems to be little all furniture was removed except for two chairs. The doors
consistency in the experimental protocols used to test these to the other rooms were closed. During the experiments,
repellents. Consequently, there are major differences in the air conditioning and the lights were switched off. The
concentration, application, and test methods (i.e., hand-in- average indoor temperature at the time of the experiments
box vs field trials) used to determine bioactivity of natural was 24.4 ± 0.8º C with a relative humidity of 74-75%.
repellents (Sukumar et al. 1991). Moreover, in the case of At 18:00 local time, 200 Ae. aegypti females were
citronella oil, counter claims of reduced or absent repellent released in the sealed corridor and allowed to disperse for
activity have been made (Lindsay et al. 1996, Centers et al. 30 min. Prior to the mosquito release, a Coleman Mosquito
2002). Deleto diffuser (model# 2950-602) containing one of four
This study was conducted to help resolve some of cartridges, or one of three candles (Table 1) was placed
these issues and determine the repellent activity of three on a chair in the center of each experimental room and
commonly used natural repellents (citronella, linalool, turned on or lit. On the other chair in a corner of the room
and geraniol) in candle or diffuser form, both indoors approximately 2 m from the diffuser or candle, opposite the
and outdoors. To ensure quality control, the current study door, a volunteer with exposed legs sat on a chair prepared
adheres to the above-mentioned EPA guidelines, as well as to collect landing mosquitoes. At 18:30, the doors to each
to currently accepted standards for testing insect repellents of the experimental rooms were opened. A head-mounted
(Govere and Durrheim 2007, Barnard et al. 2007). light covered with red foil was used to improve vision while
volunteers collected mosquitoes with an aspirator. After the
MATERIALS AND METHODS experiment, the mosquitoes in the aspirators were counted
and the remaining mosquitoes in the corridor and rooms
The repellency rates of candles and/or diffusers were removed the following morning with an entomological
containing a specific type and concentration of active net and a vacuum cleaner.
ingredient (Table 1) were determined in both an indoor and On the first day of testing, the diffusers or candles
outdoor setting. were randomly assigned to the volunteers. The candle and
diffuser experiments were conducted for three h on nine
Indoor experiments consecutive nights; nine nights of candles and nine nights of
Aedes aegypti mosquitoes were bred under insectarium diffusers for a total of 18 nights. To avoid sex and locational
conditions following the recommendations of the EPA. Ae. bias, the volunteers and diffusers or candles were rotated to
aegypti was used in evening experiments since this species different rooms on successive nights to allow each volunteer
is active in the evening in Puerto Rico as well as in Israel to test each set-up.
(Muller and Schlein, unpublished data). There are reports
on a third peak of indoor biting activity around 18:00 h Outdoor experiments
in Trinidad, parts of Africa, and Indonesia (Chadee and A readily available residential outdoor mosquito trap
Martinez 2000). Larvae were reared at 27±1º C, relative was used in lieu of volunteers to determine the effectiveness
humidity 80 ±10%, and photoperiod 16:8 hours (light:dark). of the diffuser repellents outdoors. The model chosen was
Adults were 8 days old, fed 10% sucrose, and received no the Lentek MK01 mosquito trap, sold in the U.S., which
blood meals before the test. Before the experiments, the uses 120V AC electric power and generates CO2, heat, and
insects were starved for 24 h. After each test the mosquitoes moisture by burning propane.
were discarded. Tests were performed in the tropical area of Puerto
Four authors of this study (♂♂, ♀♀) served as the Rico at the northwestern coastal plain during early to mid-
volunteer subjects and were therefore fully informed of March near the city of Mayaguez. The test site was along
the nature and purposes of the test and of any reasonably a canal surrounded by pasture and farm land. A slow, but
foreseeable physical and mental health consequences. repeatedly changing, airflow was observed during the
For the trials, the exposed legs (from knee to ankle) of balance of the test period. Experiments were conducted
each volunteer were used as a test area. The skin outside the during the dry season and consequently no rainfall
test area was covered with regular clothes to protect it from occurred. The average temperature at night was 21.5±0.9º
mosquito bites. Volunteers wore short trousers and long- C with a relative humidity of 70-75%. No dramatic weather
sleeved shirts. Immediately before each trial, the exposed changes occurred during the trial period.
skin on each volunteer was cleaned with 70% isopropyl To determine the effective range of the repellents, two
alcohol. The volunteers were advised to avoid alcohol, experiments were conducted in which four diffusers hung
caffeine, and fragrance products (e.g., perfume, cologne, on tripods (approximately 80 cm from the ground) were
hair spray, lotion, etc.) during the entire test period. arranged in squares (3 m and 6 m squares) with an MK01
Experiments were performed on the northern coastal trap placed in the middle. The diffusers were equipped
plain of Puerto Rico, during March 2004, south of the city either with citronella, linalool, geraniol, or empty cartridges.
of San Sebastian. The test site consisted of a dormitory Diffuser/trap squares were distanced 100 m apart and were
building in a church retreat. Four rooms of the same size positioned alternately (active ingredient vs empty cartridge).
and shape, with a single screened window, were situated The squares were rotated on consecutive nights to account
near each other along a corridor. From these four rooms, for locational bias.
4 Journal of Vector Ecology June 2009

Diffusers and traps were operated from one h before p<0.0001. Among the diffusers, the geraniol and linalool
average sunset (18:32) to one h after average sunrise (06:40). equipped units provided significantly more protection than
There were 12 repetitions on consecutive nights. Traps were citronella equipped units (Tukey-Kramer Test p<0.05).
emptied and mosquitoes counted daily. The field trial test Overall, however, the geraniol diffuser was about twice
protocol and site selection were in accordance with EPA as effective as the linalool diffuser and about five times as
published guidelines (Govere and Durrheim 2007, Barnard effective as the citronella diffuser in protecting a person
et al. 2007). from Ae. aegypti indoors.
Of the 1,800 female mosquitoes released for the indoor
Statistical analyses diffuser experiment (200 at each of nine repetitions), 1,103
Statistical analysis was carried out using the GraphPad (61%) tried to feed on the unprotected person (control). Of
Prism 4.0 statistical package. The numbers of mosquitoes all the released mosquitoes, 13% did not try to bite any of the
in the rooms used in indoor tests were analyzed using volunteers and were removed in the following mornings.
multivariate ANOVA technique with the mean as within- Near a citronella diffuser there were 355 feeding
subject variable. Experiments with diffusers and candles attempts, near a linalool diffuser 74 attempts, and near
were analyzed separately. Following a significant F score, a geraniol diffuser 38 attempts. When the number of
post-hoc tests (Tukey’s Honestly Significant Difference) were mosquitoes collected by the non-protected control
used to further distinguish between groups. All differences volunteer was compared to the number collected by the
were considered statistically significant if p<0.05. protected volunteers, it was found that the repellency rate
The number of mosquitoes in the traps used in outdoor of citronella diffusers was 68%, linalool diffusers 93%, and
tests were analyzed using the ANOVA technique. Following geraniol diffusers 97%.
a significant F score, the Tukey’s Honestly Significant Geraniol and citronella candles also provided
Difference post-hoc test was used to further distinguish significant protection to volunteers compared to controls
between groups. All differences were considered statistically (Figure 2) F(3,6) = 101.9, p<0.0001. Overall, however, the
significant if p < 0.05. geraniol candle provided significantly more protection
The total number of mosquitoes caught in traps and than the citronella candle, (Tukey-Kramer Test p<0.05) the
aspirators were converted to mean ± standard deviation. geraniol being about five times as effective as the citronella
Mean totals were converted to percentage of the unprotected in protecting a person from Ae. aegypti feeding indoors.
controls by using the following equations: for indoor Of the 1,800 female mosquitoes released for the indoor
experiments, the repellency rate (%) = [l - (biting attempts candle experiment (200 released at each of nine repetitions),
treated / biting attempts control)] x 100. For outdoor 908 (50%) tried to feed on the unprotected volunteer. Of all
experiments, the repellency rate (%) = [l - (total caught the released mosquitoes, 14% did not try to bite any of the
treated / total caught control)] x 100. volunteers and were removed in the following mornings.
Near a citronella candle there were 533 feeding attempts
RESULTS and near a geraniol candle there were 106 attempts. When the
number of mosquitoes caught by the non-protected control
After each overnight use, candles and diffusers were volunteer was compared to the number of mosquitoes
weighed to determine the average volume of material caught by the protected volunteers, it was found that the
expended. The volume expended was the same among the repellency rate of citronella was 41% while the repellency
candles, and the same among diffuser cartridges, regardless rate of geraniol was 88%. Commercially available geraniol
of composition. The 100% active ingredient diffusers lost and citronella were more effective in protecting volunteers
an average total weight of only 0.1 g per h, whereas the 5% in diffuser form than in candle form.
candles lost an average of 2.2 g per h (Table 1). The candles
are only 5% and therefore contain 0.05 g of active ingredient Outdoor experiments
per 1.0 g of candle wax but lose more volume than the The following common mosquito species were
diffusers. Assuming that the active ingredient is uniformly encountered in the study: Culex nigripalpus, Ae. aegypti, Ae.
distributed throughout the candle wax, the candles would mediovittatus, and Ae. sollicitans. Most of the mosquitoes
have released 0.11 g of active ingredient per h, which is caught in the 3 m2 and 6 m2 experiments were Aedes species;
nearly the same as the amount of active ingredient released the rest were Culex. There was no significant difference in
by diffusers (0.1 g). the response of the two genera towards linalool or geraniol.
Geraniol, linalool, and citronella in combination
Indoor experiments with the Coleman Mosquito Deleto Diffuser significantly
Within the first hour, about 90% of the mosquitoes, reduced mosquito capture (Figure 3; 3 m2 F(3,9) = 16.2 p<
which were ready to feed, entered one of the rooms with 0.0001; 6 m2 F(3,44) = 39.3, p < 0.0001). When arranged in a
the volunteers. During the following two hours, very little 3x3 m square, geraniol reduced the number of mosquitoes
movement was observed. After three hours, there were no caught by the MK01 mosquito trap by 95.5%, linalool by
more feeding attempts. On average, geraniol, linalool, and 88.4%, and citronella by 65.6%. When arranged in a 6x6 m
citronella diffusers all provided significant protection to square, geraniol reduced the number of mosquitoes caught
volunteers compared to controls (Figure 1) F(4,5) = 645.6, by the MK01 mosquito trap by 75%, linalool by 58%, and
Table 1. Details of equipment used in experiments. All batteries, cartridges, and candles were changed daily.

Vol. 34, no. 1

Loss of Active
Active Ingredient and Concentration Weight Amount Active Total loss of Composition of Active
Ingredient per Supplier
Release Type (%) (g) Ingredient (g/g) Weight per h (g) Ingredient
h (g)

1
Citronella** Diffuser 100% 20g 1.0 0.1 0.1 23.18% Geraniol Nature's Alkamy
34.79% Citronellal
11.19% Citronellol
0.72% Linalool
30.12% All Other*
1
Linalool Diffuser 100% 20g 1.0 0.1 0.1 95.54% Linalool Coleman
4.6% Isomers
2
Geraniol Diffuser 100% 20g 1.0 0.1 0.1 95.54% Geraniol Fasst Products
4.6% Isomers
Control Diffuser 0% n/a 0.0 n/a n/a n/a Coleman
Citronella** Candle 5% 88g 0.05 2.2 0.11 23.18% Geraniol Wal-Mart
34.79% Citronellal
11.19% Citronellol
Journal of Vector Ecology

0.72% Linalool
30.12% All Other*
Geraniol Candle 5% 88g 0.05 2.2 0.11 95.54% Geraniol Fasst Products
4.6% Isomers
Control Candle 0% 88g 0.0 n/a n/a n/a Wal-mart

*Citronella oil is comprised of more than 80 closely related terpenes, alcohols, and aldehydes (PMRA 2007). Only relevent components are listed here.
** Type "Java" from Cymbopogon winterianus.
n/a: not applicable.
1
Oil.
2
Impregnated low density polyethelene pellets.
5
6 Journal of Vector Ecology June 2009

130
120
110
100

# Mosquitoes
90
80
70
60
50
40
30

*
20
10
0
ol

or

ol

la
io

el
tr

lo
rid

an
on

on
na
or

er
C

itr
Li
C

C
Figure 1. Average number of mosquitoes caught after indoor diffuser
experiments (N=9) ± standard deviation. All repellents were significantly more
repellent than the control (F(4,5) = 645.6, p < 0.0001). *Geraniol significantly
more repellent than all others (Tukey-Kramer Test p<0.05).

120
110
100
90
# Mosquitoes

80
70
60
50
40
30
20
10 *
0
ol

or

la
io

el
tr

rid

an
on

on
or

er
C

itr
C

Figure 2. Average number of mosquitoes caught after indoor candle


experiments (N=9) ± standard deviation. All repellents were significantly
more repellent than control (F(3,6) = 101.9, p < 0.0001). *Geraniol significantly
more repellent than all others (Tukey-Kramer Test p<0.05).
Vol. 34, no. 1 Journal of Vector Ecology 7

250

# of female mosquitoes
200

150

100

50

0
itr ol

itr ol
Li la

er l

Li la

er l

l
o

io

io
el

el
tr

tr
lo

lo
an

an
on

on
on

on
na

na
C

C
G

G
C

C
3 mft square
20 square 6 mft square
10 square

Figure 3. Average number of female mosquitoes caught in MK01 mosquito


traps (N=12) ± standard deviation. Active compounds were significantly
more repellent than control (3 m square: F(3,44) = 39.3, p<0.0001) (6 m
square: F(3,9) = 16.2 p<0.0001).

citronella by 22%. All compounds provided significantly (Govere and Durrheim 2007). However, citronella diffusers
more protection (Tukey–Kramer Test p > 0.05) when placed when used indoors are fairly repellent, possibly because
closer to the MK01 trap. airflow indoors is minimal and the repellent molecules are
not diluted by airflow (i.e., building up vapor pressure).
DISCUSSION Human forearm tests of topically applied linalool report
a 91.7% repellency rate towards Cx. pipiens (Park et al. 2005).
In a previous study (Lindsay et al. 1996), 3% citronella No linalool candles were available for evaluation but we
candles and 5% citronella incense in an outdoor setting report here that linalool diffusers provided a 93% repellency
increased the repellency rate over the unprotected control rate indoors and a repellency rate of 58% outdoors. These
by only 42.3 and 24.2%, respectively. In this study, similar rates, especially indoors, support the claim that linalool
results were obtained; a 5% citronella candle increased repels mosquitoes (Govere and Durrheim 2007).
the repellency rate indoors by only 14%. However, the Studies conducted on the repellent effect of geraniol on
continuous release citronella diffusers increased the mosquitoes are limited and use varying measurements of
repellency rate indoors by 68%. Since the amount of active repellency. One laboratory study claims that 25% geraniol
ingredient expended by candles and diffusers is nearly applied topically repels mosquitoes for 3.2 to 4.8 h (Barnard
identical, one possible explanation for this difference is and Xue 2004) and still others claim geraniol repels termites
that the citronella candle releases its compounds with heat (Blaske and Hertel 2001), human body lice (Mumcuoglu et
from the flame and this may cause the active ingredients al. 1996) and some species of flies (McGuire 1984). Under
to concentrate and move vertically away from the candle the uniform conditions of our experiments, geraniol
with the heat. The diffuser, on the other hand, continuously repelled significantly more mosquitoes than citronella or
disperses an active ingredient horizontally. Only diffusers, linalool, both indoors (97% repellency with diffusers; 50%
not candles, were tested for repellency outdoors; the results with candles) and outdoors (95.5% repellency with diffusers
of which, for citronella, were poor (21%). at 3 m, 75% repellency with diffusers at 6 m).
Citronella in candle form used indoors, or in diffuser According to the data collected in the present study,
form used outdoors, does not reduce mosquitoes or mosquito geraniol diffusers, used indoors, fit the repellency criteria
bites to a degree acceptable to the EPA or conventional outlined by Govere and Durrheim (2007) and Barnard and
recommended standards for repellent testing (Govere and colleagues (2007), and also strongly suggest that geraniol
Durrheim 2007, Barnard et al. 2007). Currently accepted provides much greater indoor and outdoor protection
guidelines require candles, coils, vaporizing mats, or other coverage than the other tested products (Figures 1 and 2).
such products to provide at least a 50% repellency rate to When diffusers were tested outdoors, all compounds
make a reliable claim that the product repels mosquitoes repelled better when placed closer to the traps (Figure 3),
8 Journal of Vector Ecology June 2009

suggesting the repellent activity of botanical products can Hayes, W.J. and E.R. Laws Jr. (eds.) 1991. Handbook of
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