Insects 11 00198 v2
Insects 11 00198 v2
Insects 11 00198 v2
Article
Larvicidal and Repellent Activity of Mentha arvensis L.
Essential Oil against Aedes aegypti
Ho Dung Manh 1, * and Ong Thi Tuyet 2
1 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Lac Hong University, Dong Nai 810000,
Vietnam
2 Department of Traditional Medicine, Faculty of Pharmacy, Lac Hong University, Dong Nai 810000, Vietnam;
tuyetong2610@gmail.com
* Correspondence: manhho@lhu.edu.vn
Received: 5 March 2020; Accepted: 21 March 2020; Published: 22 March 2020
Abstract: Dengue is one of the most dangerous vector-borne diseases transmitted by Aedes mosquitoes.
The use of mosquito repellents to protect human hosts and insecticides to reduce the mosquito
population is a crucial strategy to prevent the disease. Here, we reported larvicidal and repellent
activities of Mentha arvensis L. essential oil against Aedes aegypti, the main vector of the disease.
The essential oil was extracted by hydro-distillation from the aromatic plant grown in Vietnam.
The yield was 0.67% based on the weight of fresh leaves. The essential oil was analyzed by gas
chromatography-mass spectrometry (GC-MS). The main components were menthol (66.04%), menthyl
acetate (22.19%), menthone (2.51%), and limonene (2.04%). Toxicity test on Aedes aegypti larvae
showed that the median lethal concentrations, LC50 and LC90 were 78.1 ppm (part per million)
and 125.7 ppm, respectively. Besides, the essential oil showed excellent repellency on Aedes aegypti
mosquitoes. At 25%, 50%, and 100% concentration, the respective complete protection times (CPTs)
were 45 min, 90 min, and 165 min. When adding 5% vanillin to the essential oil (25%), the complete
protection time of the essential oil increased up to 120 min. In conclusion, the EO from Mentha arvensis
L. has been shown to be a promising natural larvicide and repellent against Aedes aegypti mosquitoes.
Keywords: Mentha arvensis; essential oil; Aedes aegypti; Larvicidal activity; mosquito repellent
1. Introduction
Dengue is one of the most important vector-borne diseases and is transmitted by Aedes mosquitoes.
There are millions of infections that occur every year in the world [1,2]. Global warming and human
population growth have led to an increase in mosquito population and number of infections [3].
Disease control in practice usually includes using insecticide to reduce the mosquito population [4]
and using chemical repellents to protect the human host from mosquito bites [5].
Although synthetic insecticides such as organophosphate, pyrethroid, etc. have reduced the
Aedes mosquito population successfully, the continuous increase in the use of the synthetic insecticides
has led to mosquito resistance [6], and more importantly, potential toxicity in the environment
and adverse effects on human health [7,8]. Besides, repellents are often used to protect against
mosquito bites by applying on human skin. One of the most effective chemical repellents is DEET
(N,N-diethyl-3-methylbenzamide), which is widely used in commercial products [9]. However, DEET
has some potential risks for human health, especially a high level of DEET was reported to have
adverse effects on children [10].
Many recent studies have been focused on plant-based products that present low toxicity and
reduce the accumulation of toxic chemicals in the environment [11,12]. Among these natural products,
essential oils have been shown to be potential alternatives to synthetic chemicals because they are
effective, eco-friendly and available to many parts in the world affected by mosquito vector-borne
disease [13]. For example, common ingredients used in mosquito repellents are citronella oil, lemongrass
oil, and para-menthane 3,8-diol (PMD) [14] found in waste distillate after extraction of the eucalyptus
citriodora oil.
In this study, we aim to extract the essential oil from the Mentha arvensis L. aromatic plant, grown
in southern Vietnam, and to evaluate the larvicidal and repellent activities of the essential oil against
Aedes aegypti mosquitoes.
2.2. GC/MS
The essential oil was characterized and quantified by GC–MS analysis on an Agilent 6890N gas
chromatograph instrument equipped with an Agilent 5973 mass spectrometer and an HP-5MS capillary
column (length 30 m × 0.25 mm ID, film thickness 0.25 mm; Agilent-Technologies, Palo Alto, CA,
USA). The carrier gas was helium at a constant flow of 1.0 mL/min. The oven temperature programs
were as follows: from 50 ◦ C (held for 2 min) to 80 ◦ C (2 ◦ C/min), from 80 ◦ C to 150 ◦ C (5 ◦ C/min),
from 150 ◦ C to 200 ◦ C (10 ◦ C/min), from 200 ◦ C to 300 ◦ C (20 ◦ C/ min) and held there for 5 min. The
temperature of injector was 250 ◦ C. The samples were diluted in hexane (1:40 v/v), then 1 µL of the
diluted samples were injected in splitless mode. Component identification was done based on MS
library search (NIST and Wiley). The percentage composition was calculated by integrating the peak
areas of the chromatograms.
Table 2. Larvicidal activity of Mentha arvensis L. essential oil against Aedes aegypti larvae after 24
h exposure.
In a recent review, Pavela [23] has found that most essential oils with the value of LC50 less than
100 ppm against mosquito larvae were from five botanical families: Myrtaceae, Apiaceae, Rutaceae,
Cupressaceae, and Lamiaceae. The genus Mentha, which belongs to Lamiaceae family, is cultivated
over the world, and their oils have shown larvicidal effects on Aedes aegypti larvae [24–26]. However,
the studies on the larvicidal effect of Mentha genus against Aedes aegypti larvae was limited to Mentha
piperita [24], Mentha x villosa [25], and Mentha spicata [26]. In these previous studies, the LC50 value
of Mentha piperita oil was 98.7 ppm [24], while the Mentha spicata oil and Mentha x villosa oil showed
lower LC50 values of 56.1 ppm, 45.0 ppm, respectively. In this study, the LC50 value of Mentha arvensis
essential oil was 78.1 ppm. In comparison with the essential oils, the synthetic chemical Temephos
had a much lower LC50 value of 0.043 ppm [25]. Although Mentha arvensis L. essential oil shows the
larvicidal effect against Aedes aegypti, further investigations on field application are neccessary.
Regarding the mode of action, essential oils can produce neurotoxic effects on insects through
several targets such as inhibiting acetylcholinesterase enzyme in the cholinergic system [27] or acting on
Octopamine receptor [28,29] and GABA receptor [30]. Furthermore, the synergistic effects of essential
oil components probably increase the toxicity of the essential oil. Hummelbrunner et al. [31] found that
thymol acted synergistically with trans-anethole to enhance the acute toxicity to tobacco cutworms.
Osanlo et al. [32] found that the clove oil was more toxic than its major component eugenol. They
Insects 2020, 11, 198 5 of 9
suggested that the minor compound might act as a synergist that enhance the toxicity of the major
compound. Santos et al. [33] reported that menthol had an LC50 value of 404 ppm against Aedes aegypti
larvae. This value was much higher than LC50 value of 78.1 ppm of the Mentha arvensis essential oil,
which contained menthol as the major component in this study.
Protectiontimes
Figure1.1.Protection
Figure Menthaarvensis
timesofofMentha arvensisL.L.essential
essentialoil
oilagainst Aedesaegypti
againstAedes aegyptimosquitoes.
mosquitoes.Data
Data
are shown as mean ± SD (n = 4). Letters are used to show statistical significance (p < 0.05).
are shown as mean ± SD (n = 4). Letters are used to show statistical significance (p < 0.05). Bars are Bars are
significantly different if they do not share letters.
significantly different if they do not share letters.
The use of essential oils such as citronella oil, lemongrass oil, and eucalyptus oil is widely accepted
The use of essential oils such as citronella oil, lemongrass oil, and eucalyptus oil is widely
as mosquito repellents [11]. Mosquitoes locate the human host by integrating olfactory, thermal, and
accepted as mosquito repellents [11]. Mosquitoes locate the human host by integrating olfactory,
visual cues. Among these, odors play an important role in human host detection [35]. Mosquitoes
thermal, and visual cues. Among these, odors play an important role in human host detection [35].
detect human odors such as CO , acid lactic, and 1-octen-3-ol when that volatile odors bind to their
Mosquitoes detect human odors 2such as CO2, acid lactic, and 1-octen-3-ol when that volatile odors
odor receptors. The mode of action of mosquito repellents remains a controversial topic in which they
bind to their odor receptors. The mode of action of mosquito repellents remains a controversial topic
may activate receptors associated with repellency or inhibit receptors associated with attraction [36].
in which they may activate receptors associated with repellency or inhibit receptors associated with
Essential oils might produce effects through interaction with odor receptors, therefore decreases
attraction [36].
contacts between mosquitoes and their hosts. However, due to their high volatility, most essential oils
have short repellent action, as compared with synthetic DEET [37]. To solve this problem, Khan et
al. [38] used vanillin as a fixative to increase their protection time. Table 3 shows a review of the previous
Insects 2020, 11, 198 6 of 9
studies that reported the protection time of essential oils with and without vanillin. Tawasin et al. [39]
found that an addition of 5% vanillin to citronella oil could significantly increase the protection time
against Aedes aegypti up to 6.5 h. Songkro et al. [40] compared the evaporation rate of the citronella oil
with and without vanillin. They found that citronella oil with vanillin had a lower rate of evaporation.
Adding vanillin to Zanthoxylum piperitum [41], Curcuma longa, Eucalyptus globulus, Citrus aurantium [42],
and lemongrass oil [43], etc was also reported to increase their protection times. However, if only 5%
vanillin solution was tested, the protection time was 15 min [43]. Therefore, vanillin has acted as a
synergist to those essential oils to enhance their repellency. In the present study, we found that the
addition of 5% vanillin to the Mentha arvensis essential oil (25%) increased the protection time of the
oil nearly 3 times, from 45 min up to 120 min. Recently, nanotechnology has been applied to slow
the release rate of essential oil and thus prolong the protection time. Sakulku et al. [44] developed
citronella oil nanoemulsion and found that the release rate of the essential oil in the nanoemulsion
decreased and thus its protection time increased. Nuchuchua et al. [45] developed a nanoemulsion
containing citronella oil, hairy basil oil, and vetiver oil. They found that this nanoemulsion increased
the protection time up to 4.7 h. Although EO of Mentha arvensis L. shows promising repellency,
incorporating the EO and vanillin into a nanoemulsion might extend their mosquito protection time.
Table 3. A review of protection times of essential oils with or without vanillin, against Aedes
aegypti mosquitoes.
Protection Time
Vanillin
Essential Oils Volume/Oil Concentration Test Method Area/Part Treated (Without—With Ref.
Concentration
Vanillin)
Mentha arvensis 0.1 mL of 25% in ethanol 5% Arm-in- Cage 30 cm2 /forearm 45–120 (min) This study
Tumeric 0.1 mL of 25% in ethanol 5% Arm-in- Cage 30 cm2 /forearm 1.0–4.0 (h) [39]
Kaffir lime 0.1 mL of 25% in ethanol 5% Arm-in- Cage 30 cm2 /forearm 1.0–3.5 (h) [39]
Citronella 0.1 mL of 25% in ethanol 5% Arm-in- Cage 30 cm2 /forearm 3.0–6.5 (h) [39]
Hairy basil 0.1 mL of 25% in ethanol 5% Arm-in- Cage 30 cm2 /forearm 3.0–6.5 (h) [39]
Zanthoxylum piperitum 0.1 mL of pure oil 10% Arm-in- Cage 30 cm2 /forearm 1.0–2.5 (h) [41]
Curcuma longa 0.1 mL of 25% in coconut oil 5% Arm-in- Cage 30 cm2 /forearm 1.5–2.5 (h) [42]
Eucalyptus globulus 0.1 mL of 25% in coconut oil 5% Arm-in- Cage 30 cm2 /forearm 66–144 (min) [42]
Citrus aurantium 0.1 mL of 25% in coconut oil 5% Arm-in- Cage 30 cm2 /forearm 66–120 (min) [42]
Cassia oil 0.1 mL of 5% in ethanol 5% Arm-in- Cage 24 cm2 /forearm 75–135 (min) [43]
Rosemary 0.1 mL of 5% in ethanol 5% Arm-in- Cage 24 cm2 /forearm 0–52 (min) [43]
Lemon eucalyptus 0.1 mL of 5% in ethanol 5% Arm-in- Cage 24 cm2 /forearm 22.5–60.0 (min) [43]
Xanthoxylum 0.1 mL of 5% in ethanol 5% Arm-in- Cage 24 cm2 /forearm 30–60 (min) [43]
Lemongrass 0.1 mL of 5% in ethanol 5% Arm-in- Cage 24 cm2 /forearm 30–105 (min) [43]
4. Conclusions
Our findings suggest that Mentha arvensis L. essential oil exhibits toxicity to the larvae of Aedes
aegypti, which may be considered as a potential larvicide for controlling mosquito population. In
addition, the EO shows excellent repellency against Aedes aegypti mosquitoes when adding 5% vanillin.
Further studies on field applications as well as new repellent formulations based on the essential oil
are necessary.
Author Contributions: Conceptualization, H.D.M.; methodology, H.D.M.; software, O.T.T.; validation, O.T.T.
and H.D.M.; formal analysis, O.T.T.; investigation, and H.D.M.; resources, H.D.M.; data curation, O.T.T.;
writing—original draft preparation, O.T.T; writing—review and editing, H.D.M; visualization, O.T.T.; supervision,
H.D.M.; project administration, H.D.M.; funding acquisition, O.T.T. All authors have read and agree to the
published version of the manuscript.
Funding: This research was funded by Lac Hong University, grant number LHU-RF-MP-18-01-12.
Acknowledgments: The authors are very much grateful to Nguyen Thi Thanh Thuy, Institute of Applied Materials
Science at Ho Chi Minh city for their valuable help in GC-MS analysis of the essential oils.
Conflicts of Interest: The authors declare no conflict of interest.
Insects 2020, 11, 198 7 of 9
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