Acta Tropica 194 (2019) 93–99

Contents lists available at ScienceDirect

Acta Tropica
journal homepage: www.elsevier.com/locate/actatropica

The electronic song “Scary Monsters and Nice Sprites” reduces host attack T
and mating success in the dengue vector Aedes aegypti
⁎
Hamady Dienga,b, , Ching Chuin Thec, Tomomitsu Sathod, Fumio Miaked, Erida Wydiamalae,
Nur Faeza A. Kassimf, Nur Aida Hashimg, Ronald E. Morales Vargash, Noppawan P. Moralesi
a
Institute of Biodiversity and Environmental Conservation, Universiti Malaysia Sarawak, Malaysia
b
Mosquito Research and Control Unit, Grand Cayman, Cayman Islands
c
Faculty of Resource Science and Technology, Universiti Malaysia Sarawak, Kota Samarahan, Malaysia
d
Faculty of Pharmaceutical Sciences, Fukuoka University, Japan
e
Faculty of Medicine, Lambung Mangkurat University, South Kalimantan, Indonesia
f
School of Biological Sciences, Universiti Sains Malaysia, Penang, Malaysia
g
School of Food Science and Technology, Universiti Malaysia Terengganu, Kuaka Terengganu, Malaysia
h
Faculty of Tropical Medicine, Mahidol University, Thailand
i
Faculty of Science, Mahidol University, Thailand

A R T I C LE I N FO A B S T R A C T

Keywords: Sound and its reception are crucial for reproduction, survival, and population maintenance of many animals. In
Dengue vector insects, low-frequency vibrations facilitate sexual interactions, whereas noise disrupts the perception of signals
Music from conspecifics and hosts. Despite evidence that mosquitoes respond to sound frequencies beyond funda-
Visitation mental ranges, including songs, and that males and females need to struggle to harmonize their flight tones, the
Host attack
behavioral impacts of music as control targets remain unexplored. In this study, we examined the effects of
Blood feeding
electronic music (Scary Monsters and Nice Sprites by Skrillex) on foraging, host attack, and sexual activities of
Copulation
the dengue vector Aedes aegypti. Adults were presented with two sound environments (music-off or music-on).
Discrepancies in visitation, blood feeding, and copulation patterns were compared between environments with
and without music. Ae. aegypti females maintained in the music-off environment initiated host visits earlier than
those in the music-on environment. They visited the host significantly less often in the music-on than the music-
off condition. Females exposed to music attacked hosts much later than their non-exposed peers. The occurrence
of blood feeding activity was lower when music was being played. Adults exposed to music copulated far less
often than their counterparts kept in an environment where there was no music. In addition to providing insight
into the auditory sensitivity of Ae. aegypti to sound, our results indicated the vulnerability of its key vectorial
capacity traits to electronic music. The observation that such music can delay host attack, reduce blood feeding,
and disrupt mating provides new avenues for the development of music-based personal protective and control
measures against Aedes-borne diseases.

1. Introduction
Despite advances in control technologies, diseases transmitted by
Aedes mosquitoes—i.e., Zika, chikungunya, and dengue—are still ser-
ious public health problems worldwide (Wilder-Smith et al., 2017; Roiz
et al., 2018). As with most mosquito-borne diseases, efforts to prevent
or reduce the incidence of Aedes-borne viral illnesses have mainly fo-
cused on the application of insecticides (Caetano and Yoneyama, 2001;
WHO, 2010; Abad-Franch et al., 2017). Although this strategy has
historically been useful in managing mosquito-borne diseases (WHO,
2014), there have been no major successes in recent decades (Rivero
et al., 2010; Corbel and N’guessan, 2013; Patterson et al., 2016). In
addition to the increased resistance to all four classes of insecticide used
to date (Dia et al., 2012; Chanda et al., 2016; David et al., 2018), these
agents have a number of other disadvantages, including biodiversity
loss and adverse effects on the health of animals (Lawler, 2017) and
humans (Peterson et al., 2006), which have led to anti-pesticide acti-
vism (Logomasini, 2004; Nicolopoulou-Stamati et al., 2016) and calls
for the development of more environmentally friendly strategies
(Patterson et al., 2016; Poulin et al., 2017).

⁎
Corresponding author at: Mosquito Research & Control Unit, 99 Red Gate Rd., P.O. Box 486, Grand Cayman KY1-1106, CAYMAN ISLANDS.
E-mail address: hamachan1@yahoo.com (H. Dieng).

https://doi.org/10.1016/j.actatropica.2019.03.027
Received 26 January 2019; Received in revised form 24 March 2019; Accepted 24 March 2019
Available online 25 March 2019
0001-706X/ © 2019 Published by Elsevier B.V.
H. Dieng, et al.
The majority of non-chemical insect pest management strategies
involve manipulation of the target organism’s behavior by direct at-
traction and repellence, or indirectly by disrupting key behavioral
traits, such as host seeking, blood feeding, and mating (Foster and
Harris, 1997; Polajnar et al., 2014). Polajnar et al. (2014) suggested
that effective containment requires incorporation of knowledge about
the target’s sensory modalities.
In mosquitoes, blood meal uptake and mating are closely related
and play key roles in both population persistence and disease occur-
rence (Dieng et al., 2018a). Blood feeding is the principal process by
which disease transmission occurs (Foster, 1995; Lehane, 2005), as it
facilitates the transfer of infectious agents to and from the host (Dieng
et al., 2018a). In dengue vectors, such agents can be transferred via
mating (Martins et al., 2012), a prerequisite for the production of
subsequent generations (Dieng et al., 2006) and increasing population
density (Dieng et al., 2018a). In fact, in addition to their strong pre-
ference for human hosts (Takken and Knols, 1999; Oliva et al., 2013),
the effectiveness of mosquitoes as disease vectors is closely related to
their multi-gonotrophism. Male mosquitoes transfer sperm to the fe-
male during mating (Helinski and Harrington, 2011; Alfonso-Parra
et al., 2016). After a single successful insemination, female mosquitoes
store sperm for their whole lifetime, using it to fertilize eggs whenever a
blood meal is ingested (Clement, 1992; Childs et al., 2016). Such
mating acts necessitate a sequence of behavioral rituals (Klowden and
Zwiebel, 2005) in which flight is an essential component (Belton, 1994;
Spitzen and Takken, 2018). In aedines, mating is initiated in
swarms—an assembly of males displaying flight patterns (Klowden and
Zwiebel, 2005) or single male–female pairing (Oliva et al., 2013) in the
vicinity of a marker that is mostly a warm-blooded organism (Anderson,
1974; Jeanson, 1985). Both males and females produce sounds through
the beating of their wings (Belton, 1994), which generates vibrations
(Alexander, 1957). For successful mating to occur, the male must har-
monize its flight tone with that of its partner using auditory sensitivity,
mediated by the antennae and Johnson’s organs (Robert, 2009; Gibson
et al., 2010). In dengue vectors, males and females possess different
fundamental frequencies (male: 600 Hz; female: 400 Hz, Cator et al.,
2009). There is evidence that both sexes can hear (Robert, 2009) and
share harmonic frequencies beyond the fundamental frequency of their
respective flight tones (Clement, 1999; Cator et al., 2009).
Although sound has been reported to play an important role in the
sexual interactions and survival of many insects (Alexander, 1957;
Polajnar et al., 2014), including mosquitoes (Clement, 1999; Robert,
2009; Cator et al., 2009), there is evidence that sound has detrimental
effects on insect acoustic signal perception, courtship, and feeding ac-
tivities. Costello and Symes (2014) reported that male tree crickets
were less likely to begin calling in the presence of road noise, which
they suggested decreases the ability of females to assess and compare
males. Lee et al. (2011) examined the feeding activity on plant hosts in
the presence of acoustic stimuli with different frequencies and intensity
levels by the aphid Myzus persicae. A broad range of frequencies
(100–10,000 Hz) appeared to effectively suppress phloem feeding in
this insect. Barton et al. (2018) investigated the direct effects of an-
thropogenic sounds on the growth of plants and their herbivorous pests,
as well as predation activity of an insect biocontrol agent. Some sounds
were shown to decrease predation rates of Coccinellidae lady beetles,
thereby indirectly increasing aphid abundance and decreasing plant
biomass. There is also evidence that incidental vibrations trigger startle
responses in many insects. Dropping a metal weight on a plant or a
surface connected to the plant has been reported to induce cessation of
activity by the Colorado potato beetle (Polajnar et al., 2014). In situa-
tions similar to those described previously by these authors, cessation of
blood feeding and flying away from human hosts are often observed
after vibrational cues emitted by strong noise or slapping close to a
blood feeding female (Dieng et al., unpublished data).
In experiments using leafhoppers and planthoppers, Saxena and
Kumar (1980) reported that sound vibrations generated by a
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Acta Tropica 194 (2019) 93–99
harmonium (a keyboard instrument) or an audio-oscillator (an instru-
ment that produces radio frequency signals) interrupted acoustic and
mating communications of insects and suggested that music has the
potential to disrupt mating. Both leafhoppers and planthoppers com-
municate with their sexual partners by sound vibrations (Gillham,
1992; Eriksson et al., 2011). Although mosquitoes, including dengue
vectors, have the most acute sense of hearing known among in-
vertebrates (Göpfert and Robert, 2000) and also use acoustic vibrations
for sexual recognition and mating (Robert, 2009; Vigoder et al., 2013),
the role of music-derived vibrations as they affect mosquito behaviors
remains unexplored. Any nearby source of vibrations resonating at
frequencies different to the harmonic frequencies needed by male and
female mosquitoes to listen to each other via adjustment of their flight
tones (Robert, 2009) may alter sexual recognition and mating success.
Although it has been reported that dengue mosquitoes can create
and listen to love songs (Cator et al., 2009; Robert, 2009), this is typical
of humans (Feng, 2012), who throughout history have produced a di-
versity of musical genres (Ahrendt, 2006). Despite of differences in
pitch, rhythm, and dynamics of genres, one common factor is that
music consists of a mixture of sounds with discrete and rational fre-
quencies with a discernible dominant frequency (Nisha and Sorupani,
2018). Although music is uniquely human (Feng, 2012), devices for
playing music are increasingly been used in insect studies (Saxena and
Kumar, 1980; Ikeshoji and Yap, 1990; Mankin, 2012; Barton et al.,
2018). Recently, it has been shown that rock music can alter inter-
specific interactions and cause negative effects on insect feeding ac-
tivity (Barton et al., 2018). Despite evidence that dengue mosquitoes
have an acute sense of hearing (Göpfert and Robert, 2000), are re-
sponsive to sound frequencies beyond the fundamental range (Ikeshoji
and Yap, 1990; Cator et al., 2009), and that both sexes exert effort to
hear each other (Robert, 2009), whether music has behavioral effects
on these insects remains unknown. The present study was performed to
examine whether electronic music influences the foraging activity,
blood feeding and mating successes of the dengue vector, Aedes aegypti.
2. Materials and methods
2.1. Colony maintenance and experimental mosquitoes
The Ae. aegypti strain used in this study originated from a colony
established in 2017 at the Entomology Unit of the Faculty of Resource
Science and Technology (University Malaysia Sarawak, Kota
Samarahan) under controlled environmental conditions (temperature
26 °C–28 °C; relative humidity 75%–85%; and light:dark photoperiod of
14:10 h). Samples of eggs were flooded in aged tap water for 24 h and
the resulting newly hatched larvae (NHLs) were reared at densities
ranging from 50 to 100 per plastic tray containing 1.2 L of tap water (As
One Corporation, Osaka, Japan). They were supplied with diet con-
sisting of cat food pellets as follows: one pellet was supplied to NHLs in
powdered form by spreading onto the water surface; developing larvae
were provided two pellets every 2 days. When necessary, the rearing
medium was replaced once before completion of larval development.
Pupae were collected in small plastic vials, which were transferred into
mosquito cages. Emerging adults had permanent access to a sugar diet
(10% of sucrose solution). To produce experimental adults, pupated
individuals were singly placed into 1.5-μl of Eppendorf tubes and upon
emergence the gender of adults was determined under a stereomicro-
scope (SZ-LED; Kenis, Osaka, Japan). Males and females were placed in
two separate cages (30 × 30 × 30 cm, BugDorm; MegaView Science
Co, Ltd, Taichung, Taiwan) labeled “virgin male cage” and “virgin fe-
male cage,” respectively, to avoid any mating events prior to bioassays
requiring virgin adults. For convenience, “virgin males” and “virgin
females” are referred to as VMs and VFs, respectively.
H. Dieng, et al. Acta Tropica 194 (2019) 93–99

2.4. Testing

To examine whether the music had an impact on the behaviors of

Fig. 1. Audio file of the song “Scary monsters and nice spirits” used in the study.
2.2. Characteristics of the experimental song
Electronic music was used in this study and the song “Scary Monsters
and Nice Spirit” (duration, 4 min 5 s) by the American electronic music
producer Skrillex was selected. The dynamics of the song were analyzed
using a digital audio workstation (FL Studio 20; Image-Line, Gent,
Belgium). The song was characterized as noisy based on the resulting
vibragram and strong sound pressure/vibration with constantly rising
pitches (Fig. 1).
2.3. Experimental features
Ae. aegypti, 10 VFs that had been starved for 12 h were placed in the OA
containing one virgin conspecific male (VM) and a restrained hamster.
Immediately upon collective release of the VFs, an assistant turned the
player on to play the song and an observer began recording the time to
first visit and feeding attempt, number of visits, feeding events, and
number of copulas within 10 min. On a different day, another OA was
set up with the same experimental features, procedures, and data col-
lection process as outlined above, but with no music being played
(music-off, control). Ten additional replicates of the treatment [one OA
with 10 VFs + 1 VM + restrained hamster + music-on] and eleven
other replicates of the control [one OA with 10
VFs + 1 VM + restrained hamster + music-off] were set up on the
same or other days and monitored as described for the first OAs. All
observations were conducted during the day (10:00 and 17:00) under
controlled environmental conditions identical to those described above.
The observer was requested to avoid producing any sound during the
10-minute observation period.

All bioassays were carried out essentially according to the same

2.5. Data collection and analysis
experimental design as described previously with some modifications
(Dieng et al., 2017; Barton et al., 2018). The observation arena (OA)
In both test and control bioassays, the time to first visit attempt by
consisted of a standard mosquito cage (30 × 30 × 30 cm; BugDorm)
any one of the VFs on the mesh device was recorded immediately using
placed on a fixed virtual square with a hamster restrained within a
a stopwatch after transfer of the VFs into the cage. A visit attempt was
mesh device using round metal purse paper clips. The mesh device was
defined as a VF landing on the mesh device, as described previously
positioned at the bottom center of the cage with the clips and half
(Dieng et al., 2018b). The time between the end of the collective
covered with hard plastic material. A speaker connected to a woofer
transfer of the 10 VFs and a visit attempt was noted for each bioassay
equipped with a volume controller (2.1 Stereo speaker, Model F40;
replicate of each bioassay type (test and control). In each of these cases,
Sensonic, Johor, Malaysia) was placed close to the cage with the cone
the mean ( ± SE) of these times (expressed in seconds) was considered
facing but not touching one of the netted sides. This positioning of the
as a measure of response time to the host. Any landing event where the
speaker was done to make sound audible within the cage while
VF remained on the mesh device for at least 5 s was taken as a visit. The
avoiding any vibration effects on the cage via the mesh netting. Music
mean number of visits was calculated and used as the visitation score. A
was played using Groove Music Software 2018 version (Microsoft
VF landing on the uncovered part of the hamster and actively at-
Corp., Redmond, WA). The volume level of our music treatment
tempting to probe its skin was defined as a biting attempt, as described
(25 = 1 quarter of the maximal volume) was obtained by manipulating
previously (Dieng et al., 2017). The time between the end of release of
only the volume controller of the computer (Model A456U; AsusTek
VFs into the cage and a biting attempt was recorded for each of the
Computer Inc., Taiwan). The changes in sound pressure level during the
tested VFs in both test and control bioassays. The mean ( ± SE) of these
playing of the song analyzed using the 2nd Gen Audio Interface (model:
times (in seconds) was used as a measure of the time to first biting
Scarlett 18i20, UK) are displayed in Table 1. All observations, each
attempt. A blood feeding event was defined essentially as described
lasting for 10 min, were performed between 10:00 and 18:00 under
previously (Dieng et al., 2018b), but with slight differences: here, it was
controlled laboratory conditions (26 °C–28 °C, 75%–85% relative hu-
considered as the contact between the hamster skin and the mouthparts
midity, and 14:10 h light:dark photoperiod). For each observation,
of the VF that lasted for at least 10–20 s. The subsequent numbers of
fresh adults experimental mosquitoes starved for 12–14 h (10 VFs and
feeding events were utilized to compute the mean values. Copulation
1 VM) were used. The timing of selected mosquito female biting be-
was considered as any effective genital contact of a formed copula,
havior rituals was recorded using a stopwatch on a smartphone
which lasted for at least 10 s (Roth, 1948; Dieng et al., 2018c). The
(Huawei P9 lite; Huawei Technologies Co., Ltd., Shenzhen, Guangdong,
numbers of such sexual encounters were recorded for each replicate and
China) with lap time features and tally counters (Daiso Co., Ltd., Hir-
bioassay type, and were used to calculate the means. The discrepancies
oshima, Japan).
in response time, visitation, time to first biting attempt, blood feeding
events, and occurrence of copulation between test (music-on) and
control (music-off) conditions were compared using the non-parametric
Table 1 Mann–Whitney U test (http://astatsa.com/WilcoxonTest). In all ana-
Variations of sound pressure level (in decibels: dB) lyses, P < 0.05 was taken to indicate statistical significance.
during the playback of the song “Scary monsters and
nice spirits” at low volume.
3. Results
Time length SPL (dB)
3.1. Response times
0:00–0:30 −25 to −15
0:30–1:00 −22 to −14
1:00–1:30 −23 to −12 Ae. aegypti females showed significant variations in response time to
1:30–2:00 −24 to −15 the presence of a host related to audio player status (Mann–Whitney U
2:00–2:30 −23 to −14 test, z = 2.98, P = 0.001). Females showed a delayed response to the
2:30–3:00 −24 to −11
presence of a host when the music was playing (131.30 ± 19.04 s,
3:00–3:30 −20 to −13
3:30–4:05 −26 to −15 range 17–235 s) compared to when the player was turned off
(35.22 ± 11.06 s, range 12–115 s) (Fig. 2).

95
H. Dieng, et al.
Fig. 2. Response times by Ae. aegypti females when provided chances to take
blood meals from a vertebrate host when the audio player loaded with the
electronic music was on or turned off.
3.2. Timing of first biting attempt
The time to first biting attempt of Ae. aegypti varied appreciably
according to the audio player status (Mann–Whitney U test, z = 2.57,
P = 0.005). The time taken by females to initiate probing on the host
skin in the music-off environment was 82.44 ± 23.11 s (range
21–235 s), while the corresponding time for the music-on environment
was 191.70 ± 29.69 s (range 69–372 s) (Fig. 3).
3.3. Visitation responses
Although Ae. aegypti females visited the host regardless of whether
the music was on or off, the visitation level differed significantly be-
tween the two conditions. The mean number of visits to the host in the
music-off environment was 11.72 ± 1.29 (range 3–8) visits, while the
corresponding value when the audio player was turned on was
7.00 ± 1.19 (range 2–12) visits (Mann–Whitney U test, z = 2.14,
P = 0.015) (Fig. 4).
3.4. Patterns of blood feeding activity
Ae. aegypti females provided with an opportunity to take blood
meals from a live host fed under both conditions with different sound
Fig. 3. Mean ( ± SE) timing of first biting attempt of Ae. aegypti with reference
to the audio player status (ON or OFF) when loaded with the electronic music.
96
Acta Tropica 194 (2019) 93–99
Fig. 4. Host visitation by Ae. aegypti females with reference to the audio player
status (ON or OFF) when loaded with the electronic song.
levels (music-on and music-off conditions), but at different rates. The
mean proportion of feeding events was significantly lower in the music-
on environment compared to the music-off environment (4.00 ± 0.55
(range 2–7) vs. 6.63 ± 0.85 (range 1–11), respectively; Mann–Whitney
U test, z = 2.28, P = 0.011) (Fig. 5).
3.5. Copulation activity
Copulas were observed when Ae. aegypti females and lone males
were allowed to cohabit in both the music-off and music-on environ-
ments, but their occurrence differed between the two conditions.
Copulation was extremely rare when adults were exposed to music
(mean number of copulation events: 0.90 ± 0.40, range 0–4), while
the mean number was 5.18 ± 0.99 (range 1–12) in the music-off en-
vironment. This difference was significant (Mann–Whitney U test,
z = 3.20, P < 0.001) (Fig. 6).
Fig. 5. Mean ( ± SE) number of blood feeding events by Ae. aegypti with re-
ference to the audio player status (ON or OFF) when loaded with the electronic
music.
H. Dieng, et al.
Fig. 6. Copulation activity of Ae. aegypti in two environments with different
musical sound occurrence levels (no-sound and music being played).
4. Discussion
To our knowledge, this is the first study to test the impacts of music,
which is unique to humans (Feng, 2012), on the behaviors of a dengue
vector. Experiments using the popular electronic song “Scary Monsters
and Nice Sprites” by Skrillex using the Groove Music audio player and a
speaker indicated that playing this song near a warm-blooded verte-
brate host delayed attack, reduced blood feeding, and disrupted mating
by Ae. aegypti.
We used this electronic song in the experiments due to its loudness
and pitch, which are two factors considered to contribute to noisiness
(Nisha and Sorupani, 2018). Although we did not assess its amplitude
and frequency, the strong sound pressure/vibration and constantly
rising pitches emanating from the speaker placed close to the experi-
mental cages indicated that the song was loud and noisy.
Females exposed to such music began foraging later than their
counterparts in the music-off environment. There has been a great deal
of work regarding the effects of sound on arthropods, including insect
disease vectors. Noise is associated with detrimental effects on the
feeding activities of insects (Acheampong and Mitchell, 1997; Lee et al.,
2012). Using different music treatments at varying volumes (sound-off
and sound-on) and computer speakers, Barton et al. (2018) reported
that exposure to a rock song (“Back in Black” by AC/DC) and a mix of
urban sounds was associated with reduced feeding on prey by the lady
beetles, resulting in a marked increase in the aphid population. They
suggested that sound vibrations emanating from speakers struck the
beetles and provoked a reduction in predatory activities. Such delayed
latency to respond to nutritional sources and reduced feeding response
in the presence of noise has been reported in many other animals.
Villalobos-Jiménez et al. (2017) reported that noise decreased feeding
behavior of aquatic insects compared to quiet conditions, even when
food resources were readily available. Some groups have suggested that
noise causes stress (Simpson et al., 2015) and masks acoustic signals,
thus making detection of food difficult (Siemers and Schaub, 2011).
Female mosquitoes are bloodsucking animals (Lehane, 2005), and
bloodsucking itself represents a form of predation with humans as the
prey (Weeks and D’Antonio, 2001). In the present study, in which the
experimental procedures were similar to those reported by Barton et al.
(2018), all female mosquitoes were starved for 12–14 h, and hunger has
been often linked to instantaneous appetitive searching when a host is
present (Klowden, 1986; Lehane, 2005; Dieng et al., 2015). Following
exposure of Ae. aegypti to the electronic music, all behavioral responses
involved in host location and contact were delayed: females exposed to
music initiated foraging significantly later, visited hosts 3.72-fold later
(131.30 vs. 35.22 s, respectively) and less frequently (7 vs. 11.72 times,
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Acta Tropica 194 (2019) 93–99
respectively), and attacked the hosts much later (191.70 vs. 82.44 s,
respectively) than those maintained in the noiseless environment. In
addition, there was a clear link between the presence of sound and
blood feeding activity, with more feeding events in the silent environ-
ment than that where music was being played.
The behavior patterns in blood feeding insects, including dengue
vectors, leading to blood meal uptake are divided into the phases of
foraging (Walker and Edman, 1985) and actual feeding (Labuda and
Kozuch, 1989). Foraging consists of appetitive searching, activation/
orientation, and attraction (Lehane, 2005), whereas blood uptake in-
corporates a searching sequence (Moreira et al., 2009), stylet penetra-
tion (Ribeiro, 1987), overpowering host defenses (Walker and Edman,
1985), and seizing capacity (Clement, 1992). These behavior patterns
are not arranged hierarchically (Lehane, 2005) and when a host is
present, hunger has often been linked to the instant activation of ap-
petitive searching (Klowden, 1986; Lehane, 2005; Dieng et al., 2016).
In the present study, females tested in both the presence and absence of
music had the same starvation period, as such behavioral responses due
to differences in hunger status are unlikely. The reports cited above and
the results of the present study suggested that the sound vibrations from
the speaker during playback of the electronic music reduced stimula-
tion of the early phases of host location, i.e., appetitive searching, ac-
tivation/orientation, and detection. It is plausible that the presence of
sound vibrations and continuous pitches induced stress and fear, thus
resulting in a quiescent state in the insects. As special care was taken to
ensure that the experimental hosts were restrained with no defensive
reactions, the decreased blood feeding activity observed in the music-on
environment suggested possible interference with the probing process
by sound vibrations—e.g., exploration of suitable blood vessel, inser-
tion of stylet, or tarsal gripping ability. Epidemiologically, delayed fe-
male mosquito host attack will likely result in unsuccessful blood meal
uptake, a decreased likelihood of virus pickup or transfer, and thus
reduce the probability of disease occurrence.
In addition to biting by females during a blood meal, Aedes-borne
arboviruses that cause dengue can be transmitted to other mosquitoes
during mating (Rosen, 1987; Fontaine et al., 2018; Sánchez-Vargas
et al., 2018). Exposure to electronic music markedly reduced the co-
pulation success of Ae. aegypti. Adults maintained in an environment
with music copulated far less often than those kept without music. Si-
milar observations have been reported in many arthropods, including
insects. Saxena and Kumar (1980) exposed leafhoppers and planthop-
pers to sounds with a frequency of 200 Hz via a harmonium or an audio-
oscillator and observed interruption of their mating communication.
Therefore, they suggested that music is a potentially effective mating
disruptor. Costello and Symes (2014) reported delayed starts and re-
duced incidence of mating calls by male crickets associated with the
occurrence of road noise. They suggested that the increase in courting
latency occurred due to the inability of conspecific females to assess and
evaluate the quality of males in noisy environments. Hartbauer et al.
(2012) examined noise signal perception in crickets (Mecopoda elon-
gata) in which males exhibit synchronous mating signaling. They re-
ported that loud noise perturbed the synchrony of the male chorus, and
they attributed this effect to masking interference by the noise that
obscured conspecific signals. Similar to other reports (Brumm and
Slabbekoorn, 2005), they also suggested that noise can impede dis-
cernment of conspecific signals when there is overlap in the beat con-
tent and the intensity exceeds a critical threshold. Gordon and Uetz
(2012) investigated the effects of acoustic noise on sexual commu-
nication and mating success in an arthropod (spider). They showed that
noise affected the capacity to recognize partners, and reduced female
responsiveness to male courtship, resulting in fewer mating events and
a reduced level of mating success. Mosquitoes also use acoustic com-
munication and songs to find mates. Males must go through a range of
stereotypical actions before sperm and seminal material transfer into
the female, including pursuit, tarsal contact, ventral orientation, ter-
minalial contact, and grasping on the female (Jones and Wheeler, 1965;
H. Dieng, et al.
Klowden and Zwiebel, 2005; Ponlawat and Harrington, 2009). Most of
these behavior patterns involve flight (Belton, 1994) during which the
wing beats generate tones, referred to as songs (Arthur et al., 2014). For
mating to occur, courting partners must adjust their flight tones to
produce a common frequency (Robert, 2009; Arthur et al., 2014). Al-
though considered stereotyped, such sexual behaviors are not fixed and
can respond to other stimuli, as mosquitoes can modify their flight
tones to converge or diverge (Robert, 2009). Adult mosquitoes of the
same sex have been reported to avoid synchronizing (Vigoder et al.,
2013). In addition, the frequency range of mosquito hearing is highly
variable and can include high-frequency sound signals (2000 Hz) in
dengue mosquitoes (Cator et al., 2009). Moreover, sound impact studies
in arthropods that exhibit auditory courtship have shown that noise can
render conspecific signals ambiguous (Hartbauer et al., 2012), diminish
partner recognition ability (Gordon and Uetz, 2012), interrupt mating
communication (Saxena and Kumar, 1980), or reduce mating chances
(Costello and Symes, 2014). Any strong incidental vibration that re-
sonates at frequencies higher than those required for normal acoustic
sexual communication will likely alter the probability of mating suc-
cess. Thus, the low copulation success of Ae. aegypti observed in our
study may have been due to the interference by the electronic music
with either VM courtship performance or VF receptivity to courtship. It
is possible that some musical vibrations would have added to those of
wing beats, thus offsetting attempts to synchronize flight tone. The
vibrations emitted from the speaker may have had inhibitory effects on
tarsal contact, orientation, and seizure of VFs by the VMs. Reduced
flight activity of either VFs or lone VMs may also explain the reduced
copulation success, as acoustic maneuvers for mating generally occur in
the air. In support of this suggestion, noise has been shown to cause
quiescence in insects that use acoustic communication for their sexual
behaviors (Polajnar et al., 2014; Barton et al., 2018).
5. Conclusions
This was the first formal attempt to document the effects of music
on the behaviors of a mosquito vector. The music tested in the present
study was clearly detrimental to Ae. aegypti; females exposed to music
showed a longer response time to the mammalian host and reduced
blood feeding activity compared to their non-exposed counterparts.
Furthermore, exposure to music resulted in a very low incidence of
copulation. Thus, the electronic music acted both as an anti-mosquito
attack factor and as a mating disrupting agent. These properties suggest
the potential for development of music-based mosquito vector control
strategies. As music is loved by many people, the development of music-
based anti-mosquito control measures may represent an appealing al-
ternative to strategies involving the use of harmful chemical in-
secticides.
Acknowledgments
The authors are grateful to Rahah Binti Mohammad Yakup of the
Institute of Biodiversity and Environmental Conservation, University
Malaysia, Sarawak. This work was financially supported by the
Department of Microbiology, Faculty of Pharmaceutical Sciences,
Fukuoka University, Japan.
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