Brown marmorated stink bug (Halyomorpha halys Stl) vibration signals from a recently established invasive population in Krasnodar region of Russia

Abstract

This study was carried out because the marmorated stink bug (Halyomorpha halys) appeared in the south of Russia a few years ago and is actively expanding its habitat. The repertoire of vibration signals of marmorated stink bug from the population in Sochi, Krasnodar region of Russia was studied. It was found that the repertoire of songs partially differed from the repertoire described for populations of this species from other regions. We recorded two types of songs: the signal of males emitted in the presence of a female, and the “tremulation” before copulation. Therefore, these vibration signals registered from the marmorated stink bug from the population of the south Russia were further studied.

Full text

Res. Crop. 24 (1): 210-214 (2023)
With one figure
Printed in India
DOI: 10.31830/2348-7542.2023.ROC-11154
Brown marmorated stink bug (Halyomorpha halys Stl) vibration
signals from a recently established invasive population in
Krasnodar region of Russia
L. S. SHESTAKOV1,*
Institute for Information Transmission Problems
Russian Academy of Sciences, Moscow, 127051, Russia
*(e-mail: zicrona@yandex.ru)
(Received: November 24, 2022/Accepted: January 05, 2023)
ABSTRACT
This study was carried out because the marmorated stink bug (Halyomorpha halys)
appeared in the south of Russia a few years ago and is actively expanding its habitat.
The repertoire of vibration signals of marmorated stink bug from the population in Sochi,
Krasnodar region of Russia was studied. It was found that the repertoire of songs partially
differed from the repertoire described for populations of this species from other regions.
We recorded two types of songs: the signal of males emitted in the presence of a female,
and the “tremulation” before copulation. Therefore, these vibration signals registe red
from the marmorated stink bug from the population of the south Russia were further
studie d.
Key words: Halyomorpha halys, pest monitoring, pest population control, vibration
co mmun icat ion
1Peoples’ Friendship University of Russia, Institute of Agriculture, Moscow, 117198, Russia.
INTRODUCTION
Br o w n ma rmorated sti n k bu g
(Halyomorpha halys), which is the type species
of the genus Halyomorpha (Kment et al., 2021),
is a dangerous and economically important
polyphagous invasive pest of many crops.
Although this species originates from East Asia,
it is now actively spreading with cultivated
pl ants, show i ng ex a mples of successfu l
introduction in different regions. At the same
time, despite the quarantine measures, one
cannot speak of a slowdown in the spread, on
the contrary, the species is constantly found in
new territories. For example, in 2012, this
species entered the south of France, and already
in 2020, permanent populations were found in
the Bordeaux region (Delbac et al., 2022). The
brown marmorate d stink bug entered the
territory of Russia several years ago, and
therefore the features of its biology in the region
are poorly studied. Due to the active expansion
of this pest in the region, new signs of early
diagnostics may be required, one of which may
be communication signals.
This species can feed on more than 300
plant species and, due to its high ecological
plasticity, the ability to actively self-disperse
by flight, dispersal with cargo transportation
on plant material and the ability to mass
reproduction, can cause significant economic
and environmental damage. The species is
actively spreading in the EU countries and
harms a wide range of crops (Hess et al., 2022).
On the territory of Russia, the species was
noted in 2018 and is registe red in the
Krasnodar Territory (Mityushev, 2021; Reznik
et al., 2022). Resistant populations have been
found in several nearby countries, such as
Armenia (Kalashian et al., 2022). In order to
model the distribution and risks of invasion of
this organism into new regions, it is important
to know not only its current geographical
distribution, but also the biological features
that allo w it to actively colonize differe nt
regions and successfully develop on a wide
range of host plants. At the same time ,
theoretically, not only features and ecology, but
also morphological structures can change. So,
in a study describing the first discovery of this
species in Malta, the authors also provide
descriptions of morphological structures.
Due to the peculiarities of the biology
of this species, in some cases pest control with

insecticides leads to unsatisfactory results,
mainly due to the ability to quickly restore the
number of insects. A few authors consider the
development of long-term pest control methods
based on classical biological control strategies
to be the most promising (Giovannini et al.,
2022). Various methods of biological control of
th is spe c i e s hav e bo t h advan tag es and
disadvantages. For example, when studying
the influence of natural parasites such as the
egg parasitoid Trissolcus mitsukurii, it has
been shown that semi-natural habitats formed
by large areas of un managed ve get ation
contribute to the biological control of H. halys,
since enough parasitoi ds are maintain e d
there (Mele et al., 2022). At the same time, in
the introduce d parasitoid T. mitsukurii, the
level of parasitism on H. halys eggs was higher
th an in the lo c a l An asta tus bifasc iatus
(Hymenoptera : Eupelmidae ). However, in
orch ards this effect of hedgerows was not
significant, while a relationship was found with
distance from coastal vegetation: high levels
of T. mitsukurii parasitism were observed in
orchards close to buffer zo nes of coastal
vegetation (Mele et al., 2022).
A number of authors note that methods
based on the use of insect viromes can also
give good results. Thus, from an invasive
population from Piedmont (Italy), 7 new viral
sequences belonging to different taxonomic
groups have been described for the first time.
The presence of such a significant number of
viruses from a small population suggests a
broad association between the brown marbled
stink bug and viral entities (Giovannini et al.,
2022).
The vibrational communication of H.
halys is currently the subject of separate work,
and althoug h the repe rtoire of sig nal s is
currently well understood, there is no data on
wide geographical variability and changes at
the boundaries of the range. The repertoire of
the popu lation from the north of Italy is
described in sufficient detail (Polajnar et al.,
2016). Two type s of song s were noted in
females, and three in males. At the same time,
the authors showed that females never started
singing spontaneously and mating does not
occur if one of the partners is silent (Polajnar
et al., 2016). Male H. halys is known to
spon t an e ou s l y emi t lo n g, narrow- b a n d
vibration signals, to which nearby females
respond with their own vibration signals,
provoking exploratory behaviour in males.
During this phase, the insects emit two (in
female s) or thre e song types (in males) in
vari ou s co mbi n a tio n s un til the y make
physical contact, after which the final male
song type is characterized by tremulation.
Females never spontaneously begin to sing,
and the mating sequence does not continue if
one of the partners is silent. Male calls do not
at tract males or females, and therefo re
vi bra tions are un l ikely to play a role in
maintaining the aggregations characteristic
of this specie s, while female signals hold
pr omis e fo r d ev e lo p in g be h avi o ural
manipulation methods against this invasive
pest.
Vibration stimuli are promising for the
development of population control measures,
since the combined use of vibration signals
and aggregation pheromones as attracting
stimuli into traps is very effective, and the
opti mi z e d vi b r a t i on si g nal is th e mo s t
attractive. At the same time, when the signal
of a female was used as a stimulus, not only
th e nu mbe r of mal e s ca u ght in a tr ap
increased, but also females. Further research
is needed to evaluate the effectiveness of
vibratory traps for both early detection and
mass capture.
Signals may have more than the
obvious function of communication between
males and females. Some studies have shown
that in aggregations during wintering H. halys
exhibi t canni b alistic behavi o u r to w ard s
relatives. The depletion of metabolic reserves
and desiccation that occurs in winter can
pr ov o k e intras pe cifi c p re d a t i on . Th i s
behaviour can be facilitated not only by the
aggregation of individuals, but also by the
suppression of intraspecific signals (Papa and
Negri, 2020). In addition, H. halys have been
shown to synchronize hatching by responding
to single impulse vibrations that occur when
siblings break open eggshells. The vibration
of breaking eggs appears to be transmitted to
distant eggs in the clutch while maintaining
its signalin g functio n. It is belie v e d that
simultaneous hatching prevents cannibalism,
as larvae may eat unhatched eggs in this
species (Endo et al., 2019).
These data indicat e the promise of
studying the vibrational communication of the
brown marmorated stink bug and the need to
accumulate data on its variability in different
Brown marmorated stink bug vibration signals 211

populations. To confirm and unify effective
incentives, comparative studies of distant
geographical populations are needed, including
those at the boundaries of ranges, where
mi croe vo l u tio n a ry p r o ce s s e s are mo st
pronounced. In our study, for the first time,
we studied the repertoire of si gnal s and
reactions to vibrational stimuli in bedbugs
from a population that recently emerged in
Russia in the Krasnodar Territory.
MATERIALS AND METHODS
Insects were collected in July 2019 in
the city of Sochi, Krasnodar Territory, Russia
(about 43.57° N, 39.75° E, 5-100 m a.s.l.).
Nymphs of younger ages were kept together
in ExoTerra terrariums 20×20×30 cm. Young
twigs of laurel cherry on which insects were
collected and used as food plants. Some insects
were kept on ordinary garden cherries. Late
instar larvae were kept separately until the
imaginal molt.
Recording and Analysis of Vibrations
After molting on adults, the insects sat
down for the experiment in the following ratios:
?? (N=20), ??? (N=30) and ???? (N=30).
Signals and behaviour were recorded
simultaneously by two types of sensors: a PDV
100 laser vibrometer (Polytec, Germany) and
a GZK-661(piezocrystalline adapter) (USSR)
pickup head wi th a home made matching
amplifier. Each of the sensor types has its own
advantages. Th e technique for record i ng
signals is desc r i b e d in pre v i o u s wo rks
(She stakov , 2015). A simil ar re c o r d i ng
technique is used by other authors (Cokl et
al., 1978, Cokl, Virant-Dobelret, 2003, Polajnar
et al., 2016). The signal sampling frequency in
all cases is 44.1 kHz. This frequency is more
th an suf fi c i en t, be ca u s e th e do minan t
frequency of vibration signals is mainly in the
region of 200 Hz.
At the same time, video typing of the
behaviour was carried out on a Sony HDR-
CX405 camera. Behaviour analysis was carried
out using the free access program BORIS
(behav i o ur obs e r v ati o n system): A fr e e ,
versatile open-source event-logging software
for video/audio coding and live observations
(Friard and Gamba, 2016). According to this
program, the type of behaviour can be defined
as “state event” and “point event” (with and
without duration, respectively).
The measurement of the amplitude-
time parameters of the signals was carried out
in Cool Edit Pro 2.0 (Syntrillium Software 2001
– Fort Wayne, Indiana, USA). Signal emissions
were monitored with headphones during all
times of insect observation. Chebuchev1 and
Buttervorth filters were used to reduce the
signal/noise leve l. The sampling frequency
was 44.1 kHz. Data were statistically analyzed
in Excel 2010 and Statistica.
RESULTS AND DISCUSSION
In a l l ty p e s o f e x p er i m e nt s , we
registered only two types of songs: the signal
of males emitted in the presence of a female
and “tremulation” before copulation (see 1 to 5
in Fig. 1). The characteristics of the signals
are presented in Table 1. The first type of signal
consists of a series of pulses following each
other at approximately equal intervals. The
spect rum of the signal is noisy, wi thout
pronounced harmonics as shown 4, 5 and 7 in
Fig. 1. When emitting a signal, the insect can
move around the plant or remain motionless.
The signal was emitted only in the presence
of fe males and could prece de copul ati on
attempts. There are no visible movements of
the body now of signal emission; we assume
that it is emitted by the timbal organs.
Table 1. Characteristics of the Halyomorpha halys songs
Pu l se Pu l se Series Pulse Pu l se
dura tion pe ri od duration int er v al frequency
(ms) (ms) (s) (ms) (Hz)
M 418 . 7 753 . 55 37. 13 301. 85 172 . 19
SD 56. 76 36.79 1. 57 37.27 0. 04
M: Mean and SD: Standard deviation.
The second type of signal is emitted
immediately before copulation, and the male’s
body tremulation is noticeable. The signal is
a si n g l e vibra t i o n a l message witho u t a
pronounced division into pulses. Males, both
in pairs and in group encounters, actively
moved around the plant and tried to copulate
with females both after signal emission and
without it. At the same time, no protest signals
or response signals were found in females.
This behaviour is quite different from
that described earlier. Previous studies on
various species of bed bug s suggest that
212 Shestakov
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vibration signals are essential ele ments of
communication and an important element of
courtship behaviour. Therefore, it could be
assu med that change s in the structure of
communi c ation should greatly affe ct the
effectiveness of the interaction of the sexes.
Th i s pri n c i ple is funda me n tal in th e
development of population control measures
using insect communication signals (Polajnar
et al., 2019). The first logical suggestion why
the number of signals in the population from
the Krasnodar Territory differs greatly from that
described for the populations from Italy could
be related to the difference in the stages of
the life cycle. For example, if insects were
taken from clusters after wintering, or before
wintering. In this case, one could expect both
a decrease in the number of signals, and their
comple t e abs e nce . Ho weve r, the summer
generation obtained from larvae and ready for
reproduction was used for work. Those males
and females were fertile, did not copulate, and
must had a high motivation to form pairs. The
readiness of insects for mating was confirmed
not only by the active attempts of males to
copulate, but also by successful mating acts.
Therefore, the assumption of a difference in
the repertoire due to a possible difference in
the stage of the life cycle is not valid.
At the same time, the male signal and
“tremulation” have a similar structure to those
described in the article by Polajnar et al. (2016)
differing slightly in frequency structure. The
spectrum of signals recorded by us is noisier,
without pronounced harmonics. This can be
explained by the difference in the substrate
on which the signals were recorded, in contrast
to soybean seedlings, cherry laure l has a
denser, woody structure, the leaves are also
dense and leathery. It is known that different
substrates are a non-linear frequency filter,
which can distort the structure of the signal
in different plants (Michelsen et al., 1982;
Cokl, Virant-Dobelret, 2003). At the same
time, the amplitude-time pattern remained
practically unchange d, as did the carrier
frequency. It can be assumed that the response
signals of females are suppressed to a greater
extent on branches of laurel cherry than on
herbaceous crops. However, since the marbled
bug is polyphagous and feeds on a wide range
of herbaceous and woody plants, further
development of effective stimuli must consider
th e se diffe r e nces. Sin ce the reg istration
technique worked well in previous studies and
practically did not differ from that used by other
au tho rs, we assume tha t this opt i o n is
extremely unlikely.
The next assumption that can explain
such strong differences in the repertoire of
different populations may be since European
populations are well-established and have
been introduced into their stations for a long
time. The population in Sochi is located at the
ve ry borde r of the range and in active
movement to the north of the range, which
implies a constant search for the most suitable
food plants and the need to quickly adapt to
new conditions. In such unstable populations,
selection can go to a more active search for a
partner and the removal of “extra” patterns of
behavi our. The insects we colle cte d were
confined mainly to plantings of laurel cherry,
and according to some reports, marble bugs in
Sochi quite often infect citrus fruits. When
trying to transplant insects to other fodder
plan ts (raspb e rries, garde n che rries), all
individuals died before reaching the imaginal
molt. Therefore, we do not exclude the fact that
this population at the border of the range has
changed properties. However, this question
remains open.
CONCLUSION
Th e ma rm o r a t e d st i n k bu g is a
dangero us inva sive pe st that is actively
expanding its habitat. A number of studies
suggest that vibration songs can be a good
addition to traditional pest control methods. Our
work shows that it is necessary to take into
account differences in repertoire in different
Fig. 1. Oscillograms (1-2) and sonograms (4,5,7)
vibration songs of brown marmorated stink
bug, Halyomorpha halys Stål.
Brown marmorated stink bug vibration signals 213

geographical populations. In the future, we plan
to test the effectiveness of stimuli created on
the basis of the registered signals.
ACKNOWLEDGEMENT
This study was funded by a grant
Russian Foundation for Basic Research No. 20-
04-00553A (2020, Moscow, Russia).
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