Eared Moths Research Articles

Stealth echolocation in aerial hawking bats reflects a substrate gleaning ancestry

Predator-prey co-evolution can escalate into an evolutionary arms race.1 Examples of insect countermeasures to bat echolocation are well-known,2 but presumptive direct counter strategies in bats to insect anti-bat tactics are rare. The emission of very low-intensity calls by the hawking Barbastella barbastellus to circumvent high-frequency moth hearing is the most convincing countermeasure known.2,3 However, we demonstrate that stealth echolocation did not evolve through a high-intensity aerial hawking ancestor becoming quiet as previously hypothesized2,3,4 but from a gleaning ancestor transitioning into an obligate aerial hawker. Our ancestral state reconstructions show that the Plecotini ancestor likely gleaned prey using low-intensity calls typical of gleaning bats and that this ability-and associated traits-was subsequently lost in the barbastelle lineage. Barbastelles did not, however, revert to the oral, high-intensity call emission that other hawking bats use but retained the low-intensity nasal emission of closely related gleaning plecotines despite an extremely limited echolocation range. We further show that barbastelles continue to emit low-intensity calls even under adverse noise conditions and do not broaden the echolocation beam during the terminal buzz, unlike other vespertilionids attacking airborne prey.5,6 Together, our results suggest that barbastelles' echolocation is subject to morphological constraints prohibiting higher call amplitudes and beam broadening in the terminal buzz. We suggest that an abundance of eared prey allowed the co-opting and maintenance of low-intensity, nasal echolocation in today's obligate hawking barbastelle and that this unique foraging behavior7 persists because barbastelles remain a rare, acoustically inconspicuous predator to eared moths. VIDEO ABSTRACT.

Contrasting laboratory and field outcomes of bat-moth interactions.

Predator-prey interactions are important but difficult to study in the field. Therefore, laboratory studies are often used to examine the outcomes of predator-prey interactions. Previous laboratory studies have shown that moth hearing and ultrasound production can help prey avoid being eaten by bats. We report here that laboratory behavioural outcomes may not accurately reflect the outcomes of field bat-moth interactions. We tested the success rates of two bat species capturing moths with distinct anti-bat tactics using behavioural experiments. We compared the results with the dietary composition of field bats using next-generation DNA sequencing. Rhinolophus episcopus and Rhinolophus osgoodi had a lower rate of capture success when hunting for moths that produce anti-bat clicks than for silent eared moths and earless moths. Unexpectedly, the success rates of the bats capturing silent eared moths and earless moths did not differ significantly from each other. However, the field bats had a higher proportion of silent eared moths than that of earless moths and that of clicking moths in their diets. The difference between the proportions of silent eared moths and earless moths in the bat diets can be explained by the difference between their abundance in bat foraging habitats. These findings suggest that moth defensive tactics, bat countertactics and moth availability collectively shape the diets of insectivorous bats. This study illustrates the importance of using a combination of behavioural experiments and molecular genetic techniques to reveal the complex interactions between predators and prey in nature.

Achroia grisella’s ear as inspiration for acoustic sensing

Bio-inspiration, gathering insight from looking at nature, can be useful when trying to solve challenges innovatively. Insects are forced by their body size and the energy cost of a hearing organ to make many clever adaptations for hearing. Achroia grisella is a nocturnal moth with a hearing sense. It uses it both for avoiding predators and mating, the latter being an unconventional role among moths. The moth is capable of directionality that seems to be monoaural and dependent exclusively on morphology. We develop a model with the goal of 3D printing a structure whose behavior is similar to that of the moth ear, i.e., directionality at a low design cost. Starting from a simple circular plate model, we progressively incorporate more complex elements to improve the resemblance to the natural system. Equations, simulations, and 3D printed devices' frequency responses (measured through Laser Doppler Vibrometry) are compared. The directionality of the printed devices is analyzed.

Finding each other: Achroia grisella’s oddly simple directional ear

Achroia grisella is a pyraloid moth capable of directional hearing. It is generally accepted that hearing arose in moths to avoid bats, their main predators, and, in this particular case, was later repurposed into a rudimentary mating tool. The male emits ultrasonic signals of a wavelength several times larger than the distance between their tympana during its mating process. Notwithstanding, the female moths are capable of somewhat efficiently reaching the males after some zigzagging. Another remarkable feature is the simple structure of the ear, consisting of a cluster of just four receptor cells attached directly to the tympanic membrane. Moths with only one functioning ear have been seen to reach the males still, which implies the tympanum structure itself must confer the moth with monoaural directionality. A model is developed in COMSOL to explain the behavior of the moth ear. The complexity of the model is increased by improving the resemblance to the natural one, starting from a circular plate and progressing to, eventually, a damped elliptical plate with two different thicknesses and an attached point mass.

Light might suppress both types of sound-evoked antipredator flight in moths.

Urbanization exposes wild animals to increased levels of light, affecting particularly nocturnal animals. Artificial light at night might shift the balance of predator–prey interactions, for example, of nocturnal echolocating bats and eared moths. Moths exposed to light show less last‐ditch maneuvers in response to attacking close‐by bats. In contrast, the extent to which negative phonotaxis, moths’ first line of defense against distant bats, is affected by light is unclear. Here, we aimed to quantify the overall effect of light on both types of sound‐evoked antipredator flight, last‐ditch maneuvers and negative phonotaxis. We caught moths at two light traps, which were alternately equipped with loudspeakers that presented ultrasonic playbacks to simulate hunting bats. The light field was omnidirectional to attract moths equally from all directions. In contrast, the sound field was directional and thus, depending on the moth's approach direction, elicited either only negative phonotaxis, or negative phonotaxis and last‐ditch maneuvers. We did not observe an effect of sound playback on the number of caught moths, suggesting that light might suppress both types of antipredator flight, as either type would have caused a decline in the number of caught moths. As control, we confirmed that our playback was able to elicit evasive flight in moths in a dark flight room. Showing no effect of a treatment, however, is difficult. We discuss potential alternative explanations for our results, and call for further studies to investigate how light interferes with animal behavior.

Neural representation of bat predation risk and evasive flight in moths: A modelling approach

Most animals are at risk from multiple predators and can vary anti-predator behaviour based on the level of threat posed by each predator. Animals use sensory systems to detect predator cues, but the relationship between the tuning of sensory systems and the sensory cues related to predator threat are not well-studied at the community level. Noctuid moths have ultrasound-sensitive ears to detect the echolocation calls of predatory bats. Here, combining empirical data and mathematical modelling, we show that moth hearing is adapted to provide information about the threat posed by different sympatric bat species. First, we found that multiple characteristics related to the threat posed by bats to moths correlate with bat echolocation call frequency. Second, the frequency tuning of the most sensitive auditory receptor in noctuid moth ears provides information allowing moths to escape detection by all sympatric bats with similar safety margin distances. Third, the least sensitive auditory receptor usually responds to bat echolocation calls at a similar distance across all moth species for a given bat species. If this neuron triggers last-ditch evasive flight, it suggests that there is an ideal reaction distance for each bat species, regardless of moth size. This study shows that even a very simple sensory system can adapt to deliver information suitable for triggering appropriate defensive reactions to each predator in a multiple predator community.

Species‐specific strategies increase unpredictability of escape flight in eared moths

Abstract Many prey species overlap in time and space and are hunted by the same predators. A common anti‐predator behaviour is using evasive manoeuvres to escape an attacking predator. The escape‐tactic diversity hypothesis postulates that species‐specific differences in evasive behaviour will increase the overall unpredictability experienced by predators within a predator–prey community. Evolutionary, escape‐tactic diversity would be driven by the enhanced predator protection for each prey individual in the community. However, escape‐tactic diversity could also be a functional consequence of morphological differences that correlate with evasive capabilities. Echolocating bats and eared moths are a textbook example of predator–prey interactions. Moths exhibit evasive flight with diverse tactics; however, the variability of their evasive flight within and between species and individuals has never been quantified systematically. In addition, moth species show variation in size, which correlates with their flight capability. We recorded flight strength during tethered flight of eight sympatric moth species in response to the same level of simulated bat predation. Our method allowed us to record kinematic parameters that are correlated with evasive flight in a controlled way to investigate species‐specific differences in escape tactics. We show species‐specific and size‐independent differences in both overall flight strength and change of flight strength over time, supporting the escape‐tactic diversity hypothesis for eared moths. Additionally, we show strong interindividual differences in evasive flight within some species. This diversity in escape tactic between eared moths increases the overall unpredictability of evasive flight experienced by bat predators, likely providing increased protection against predatory bats for the single individual.

An experimental test of the allotonic frequency hypothesis to isolate the effects of light pollution on bat prey selection.

Artificial lights may be altering interactions between bats and moth prey. According to the allotonic frequency hypothesis (AFH), eared moths are generally unavailable as prey for syntonic bats (i.e., bats that use echolocation frequencies between 20 and 50kHz within the hearing range of eared moths) due to the moths' ability to detect syntonic bat echolocation. Syntonic bats therefore feed mainly on beetles, flies, true bugs, and non-eared moths. The AFH is expected to be violated around lights where eared moths are susceptible to exploitation by syntonic bats because moths' evasive strategies become less effective. The hypothesis has been tested to date almost exclusively in areas with permanent lighting, where the effects of lights on bat diets are confounded with other aspects of human habitat alteration. We undertook diet analysis in areas with short-term, localized artificial lighting to isolate the effects of artificial lighting and determine if syntonic and allotonic bats (i.e., bats that use echolocation frequencies outside the hearing range of eared moths) consumed more moths under conditions of artificial lights than in natural darkness. We found that syntonic bats increased their consumption of moth prey under experimentally lit conditions, likely owing to a reduction in the ability of eared moths to evade the bats. Eared moths may increase in diets of generalist syntonic bats foraging around artificial light sources, as opposed to allotonic species and syntonic species with a more specialized diet.

Continued source level reduction during attack in the low‐amplitude bat Barbastella barbastellus prevents moth evasive flight

Abstract Ears evolved independently in many insect taxa due to the selection pressure of echolocating bats. Eared moths perform evasive flight manoeuvres upon hearing approaching bats, thereby substantially increasing survival probability. Accordingly, eared moths constitute only a minor portion of many bats’ diets. In contrast, the barbastelle bat Barbastella barbastellus almost exclusively feeds on eared moths by emitting low‐amplitude stealth echolocation calls that are undetectable by distant moths. While closing in on the prey, however, the calls might become audible. We thus hypothesised that barbastelle bats lower the source level of their calls even further while closing in, such that the level at the moth's ear remains below the moth's hearing threshold and the moth fails to elicit its evasive manoeuvre. We tested these hypotheses by offering tethered moths to barbastelle bats in the wild and in captivity. We reconstructed the bats’ three‐dimensional flight paths based on time‐of‐arrival differences of the echolocation calls at four microphones, measured the received sound levels at the moth's position with an additional miniature microphone, and calculated call source levels. We show that barbastelle bats continuously reduced source level upon detecting a moth, thereby delaying the time and shortening the distance when the moth might detect the bat. The received level at the targeted moth never exceeded the moth's neuronal hearing threshold by more than 10 dB, which is likely sufficiently low to prevent moth evasive flight. Barbastelle bats emit low‐amplitude calls to counter moth hearing. Continued amplitude reduction during an attack extends the time before the prey becomes aware of the predator, if at all. In combination, these two strategies allowed barbastelle bats to access a food resource which is largely unavailable to competing species, thereby probably altering competition between sympatric predators. A plain language summary is available for this article.

Insect ear biomechanics: Passive and active processes that can inspire acoustic sensors

Biological systems provide an incredible wealth of archetypes that have emerged through evolutionary processes. Hearing organs are a good example of how different solutions, and adaptions, across different animal taxa, can often converge to solve similar sensory problems. Hearing has evolved independently multiple times across the insects, and the diversity of these biological solutions therefore provides a wealth of inspiration for the creation of novel acoustic sensors. Some biological solutions can be considered as purely passive mechanical constructs that accomplish some processing of the incoming sound. This talk will consider examples of this, including the frequency discrimination of the locust ear, and the directionality and wideband response of different moth ears. Several insects also display active hearing processes, whereby energy is used to actively change the hearing response, bearing some similarity to the processes found in the mammalian inner ear. This talk will thus discuss how some moths actively tune their ear, and how the mosquito ear actively utilizes gain and compression. Finally, the talk will discuss some examples of different acoustic sensors from the University of Strathclyde that take inspiration from these passive and active biological processes.

Simple Ears Inspire Frequency Agility in an Engineered Acoustic Sensor System

Standard microphones and ultrasonic devices are generally designed with a static and flat frequency response in order to address multiple acoustic applications. However, they may not be flexible or adaptable enough to deal with some requirements. For instance, when operated in noisy environments such devices may be vulnerable to wideband background noise which will require further signal processing techniques to remove it, generally relying on digital processor units. In this paper, we consider if microphones and ultrasonic devices could be designed to be sensitive only at selected frequencies of interest, whilst also providing flexibility in order to adapt to the different signals of interest and to deal with environmental demands. This research exploits the concept where the “transducer becomes part of the signal processing chain” by exploring feedback processes between mechanical and electrical mechanisms that together can enhance peripheral sound processing. This capability is present within a biological acoustic system, namely in the ears of certain moths. That was used as the model of inspiration for a smart acoustic sensor system which provides dynamic adaptation of its frequency response with amplitude and time dependence according to the input signal of interest.

Bidirectional Echolocation in the Bat Barbastella barbastellus: Different Signals of Low Source Level Are Emitted Upward through the Nose and Downward through the Mouth

The Barbastelle bat (Barbastella barbastellus) preys almost exclusively on tympanate moths. While foraging, this species alternates between two different signal types. We investigated whether these signals differ in emission direction or source level (SL) as assumed from earlier single microphone recordings. We used two different settings of a 16-microphone array to determine SL and sonar beam direction at various locations in the field. Both types of search signals had low SLs (81 and 82 dB SPL rms re 1 m) as compared to other aerial-hawking bats. These two signal types were emitted in different directions; type 1 signals were directed downward and type 2 signals upward. The angle between beam directions was approximately 70°. Barbastelle bats are able to emit signals through both the mouth and the nostrils. As mouth and nostrils are roughly perpendicular to each other, we conclude that type 1 signals are emitted through the mouth while type 2 signals and approach signals are emitted through the nose. We hypothesize that the “stealth” echolocation system of B. barbastellus is bifunctional. The more upward directed nose signals may be mainly used for search and localization of prey. Their low SL prevents an early detection by eared moths but comes at the expense of a strongly reduced detection range for the environment below the bat. The more downward directed mouth signals may have evolved to compensate for this disadvantage and may be mainly used for spatial orientation. We suggest that the possibly bifunctional echolocation system of B. barbastellus has been adapted to the selective foraging of eared moths and is an excellent example of a sophisticated sensory arms race between predator and prey.

Dynamics of the echolocation beam during prey pursuit in aerial hawking bats

In the evolutionary arms race between prey and predator, measures and countermeasures continuously evolve to increase survival on both sides. Bats and moths are prime examples. When exposed to intense ultrasound, eared moths perform dramatic escape behaviors. Vespertilionid and rhinolophid bats broaden their echolocation beam in the final stage of pursuit, presumably as a countermeasure to keep evading moths within their "acoustic field of view." In this study, we investigated if dynamic beam broadening is a general property of echolocation when catching moving prey. We recorded three species of emballonurid bats, Saccopteryx bilineata, Saccopteryx leptura, and Rhynchonycteris naso, catching airborne insects in the field. The study shows that S. bilineata and S. leptura maintain a constant beam shape during the entire prey pursuit, whereas R. naso broadens the beam by lowering the peak call frequency from 100 kHz during search and approach to 67 kHz in the buzz. Surprisingly, both Saccopteryx bats emit calls with very high energy throughout the pursuit, up to 60 times more than R. naso and Myotis daubentonii (a similar sized vespertilionid), providing them with as much, or more, peripheral "vision" than the vespertilionids, but ensonifying objects far ahead suggesting more clutter. Thus, beam broadening is not a fundamental property of the echolocation system. However, based on the results, we hypothesize that increased peripheral detection is crucial to all aerial hawking bats in the final stages of prey pursuit and speculate that beam broadening is a feature characterizing more advanced echolocation.

Unexpected dynamic up-tuning of auditory organs in day-flying moths.

In certain nocturnal moth species the frequency range of best hearing shifts to higher frequencies during repeated sound stimulation. This could provide the moths with a mechanism to better detect approaching echolocating bats. However, such a dynamic up-tuning would be of little value for day-flying moths that use intra-specific acoustic communication. Here we examined if the ears of day-flying moths provide stable tuning during longer sound stimulation. Contrary to our expectations, dynamic up-tuning was found in the ear of the day-flying species Urania boisduvalii and Empyreuma pugione. Audiograms were measured with distortion-product otoacoustic emissions (DPOAEs). The level of the dominant distortion product (i.e. 2f1-f2) varied as a function of time by as much as 45dB during ongoing acoustic stimulation, showing a systematic decrease at low frequencies and an increase at high frequencies. As a consequence, within about 2s of acoustic stimulation, the DPOAEs audiogram shifted from low to high frequencies. Despite the up-tuning, the range of best audition still fell within the frequency band of the species-specific communication signals, suggesting that intra-specific communication should not be affected adversely. Up-tuning could be an ancestral condition in moth ears that in day-flying moths does not underlie larger selection pressure.

Simple ears – flexible behavior: Information processing in the moth auditory pathway

Abstract Lepidoptera evolved tympanic ears in response to echolocating bats. Comparative studies have shown that moth ears evolved many times independently from chordotonal organs. With only 1 to 4 receptor cells, they are one of the simplest hearing organs. The small number of receptors does not imply simplicity, neither in behavior nor in the neural circuit. Behaviorally, the response to ultrasound is far from being a simple reflex. Moths’ escape behavior is modulated by a variety of cues, especially pheromones, which can alter the auditory response. Neurally the receptor cell(s) diverges onto many interneurons, enabling parallel processing and feature extraction. Ascending interneurons and sound-sensitive brain neurons innervate a neuropil in the ventrolateral protocerebrum. Further, recent electrophysiological data provides the first glimpses into how the acoustic response is modulated as well as how ultrasound influences the other senses. So far, the auditory pathway has been studied in noctuids. The findings agree well with common computational principles found in other insects. However, moth ears also show unique mechanical and neural adaptation. Here, we first describe the variety of moths’ auditory behavior, especially the co-option of ultrasonic signals for intraspecific communication. Second, we describe the current knowledge of the neural pathway gained from noctuid moths. Finally, we argue that Galleriinae which show negative and positive phonotaxis, are an interesting model species for future electrophysiological studies of the auditory pathway and multimodal sensory integration, and so are ideally suited for the study of the evolution of behavioral mechanisms given a few receptors.

Stacking the odds: light pollution may shift the balance in an ancient predator–prey arms race

Summary Artificial night lighting threatens to disrupt strongly conserved light‐dependent processes in animals and may have cascading effects on ecosystems as species interactions become altered. Insectivorous bats and their prey have been involved in a nocturnal, co‐evolutionary arms race for millions of years. Lights may interfere with anti‐bat defensive behaviours in moths, and disrupt a complex and globally ubiquitous interaction between bats and insects, ultimately leading to detrimental consequences for ecosystems on a global scale. We combined experimental and mathematical approaches to determine effects of light pollution on a free‐living bat–insect community. We compared prey selection by Cape serotine bats Neoromicia capensis in naturally unlit and artificially lit conditions using a manipulative field experiment, and developed a probabilistic model based on a suite of prey‐selection factors to explain differences in observed diet. Moth consumption by N. capensis was low under unlit conditions (mean percentage volume ± SD: 5·91 ± 6·25%), while moth consumption increased sixfold (mean percentage volume ± SD: 35·42 ± 17·90%) under lit conditions despite a decrease in relative moth abundance. Predictive prey‐selection models that included high‐efficacy estimates for eared‐moth defensive behaviour found most support given diet data for bats in unlit conditions. Conversely, models that estimated eared‐moth defensive behaviour as absent or low found more support given diet data for bats in lit conditions. Our models therefore suggest the increase in moth consumption was a result of light‐induced, decreased eared‐moth defensive behaviour. Policy implications. In the current context of unyielding growth in global light pollution, we predict that specialist moth‐eating bats and eared moths will face ever‐increasing challenges to survival through increased resource competition and predation risk, respectively. Lights should be developed to be less attractive to moths, with the goal of reducing effects on moth behaviour. Unfortunately, market preference for broad‐spectrum lighting and possible effects on other taxa make development of moth‐friendly lighting improbable. Mitigation should therefore focus on the reduction of temporal, spatial and luminance redundancy in outdoor lighting. Restriction of light inside nature reserves and urban greenbelts can help maintain dark refugia for moth‐eating bats and moths, and may become important for their persistence.

Intraspecific acoustic communication and mechanical sensitivity of the tympanal ear of the gypsy moth Lymantria dispar

AbstractTympanal ears of female gypsy moths Lymantria dispar dispar (L.) (Lepidoptera: Erebidae: Lymantriinae) are reportedly more sensitive than ears of conspecific males to sounds below 20 kHz. The hypothesis is tested that this differential sensitivity is a result of sex‐specific functional roles of sound during sexual communication, with males sending and females receiving acoustic signals. Analyses of sounds produced by flying males reveal a 33‐Hz wing beat frequency and 14‐kHz associated clicks, which remain unchanged in the presence of female sex pheromone. Females exposed to playback sounds of flying conspecific males respond with wing raising, fluttering and walking, generating distinctive visual signals that may be utilized by mate‐seeking males at close range. By contrast, females exposed to playback sounds of flying heterospecific males (Lymantria fumida Butler) do not exhibit the above behavioural responses. Laser Doppler vibrometry reveals that female tympana are particularly sensitive to frequencies in the range produced by flying conspecific males, including the 33‐Hz wing beat frequency, as well as the 7‐kHz fundamental frequency and 14‐kHz dominant frequency of associated clicks. These results support the hypothesis that the female L. dispar ear is tuned to sounds of flying conspecific males. Based on previous findings and the data of the present study, sexual communication in L. dispar appears to proceed as: (i) females emitting sex pheromone that attracts males; (ii) males flying toward calling females; and (iii) sound signals from flying males at close range inducing movement in females, which, in turn, provides visual signals that could orient males toward females.

Sensory escape from a predator–prey arms race: Low amplitude biosonar beats moth hearing

Ultrasound-sensitive ears evolved in many nocturnal insects, including some moths, to detect bat echolocation calls and evade capture. Although there is evidence that some bats emit echolocation calls that are inconspicuous to eared moths, it is difficult to determine whether this was an adaptation to moth hearing or originally evolved for a different purpose. Here we present the first example of an echolocation counterstrategy to overcome prey hearing at the cost of reduced detection distance, providing an example of a predator outcompeting its prey despite the life-dinner-principle. Aerial-hawking bats generally emit high-amplitude echolocation calls to maximize detection range. Using comparative acoustic flight-path tracking of free-flying bats, we show that the barbastelle, Barbastella barbastellus, emits calls that are 10 to 100 times lower in amplitude than those of other aerial hawking bats. Model calculations demonstrate that only bats emitting such low-amplitude calls hear moth echoes before their calls are conspicuous to moths. We confirm that the barbastelle remains undetected by moths until close and preys mainly on eared moths, using moth neurophysiology in the field and fecal DNA analysis. This adaptive stealth echolocation allows the barbastelle to access food resources that are difficult to catch for high-intensity bats.

Moth hearing and sound communication.

Active echolocation enables bats to orient and hunt the night sky for insects. As a counter-measure against the severe predation pressure many nocturnal insects have evolved ears sensitive to ultrasonic bat calls. In moths bat-detection was the principal purpose of hearing, as evidenced by comparable hearing physiology with best sensitivity in the bat echolocation range, 20-60 kHz, across moths in spite of diverse ear morphology. Some eared moths subsequently developed sound-producing organs to warn/startle/jam attacking bats and/or to communicate intraspecifically with sound. Not only the sounds for interaction with bats, but also mating signals are within the frequency range where bats echolocate, indicating that sound communication developed after hearing by "sensory exploitation". Recent findings on moth sound communication reveal that close-range (~ a few cm) communication with low-intensity ultrasounds "whispered" by males during courtship is not uncommon, contrary to the general notion of moths predominantly being silent. Sexual sound communication in moths may apply to many eared moths, perhaps even a majority. The low intensities and high frequencies explain that this was overlooked, revealing a bias towards what humans can sense, when studying (acoustic) communication in animals.

Tone-Deaf Ears in Moths May Limit the Acoustic Detection of Two-Tone Bats

Frequency alternation in the echolocation of insectivorous bats has been interpreted in relation to ranging and duty cycle, i.e. advantages for echolocation. The shifts in frequency of the calls of these so-called two-tone bats, however, may also play its role in the success of their hunting behavior for a preferred prey, the tympanate moth. How the auditory receptors (e.g. the A₁ and A₂ cells) in the moth's ear detect such frequency shifts is currently unknown. Here, we measured the auditory responses of the A₁ cell in the noctuid Spodoptera frugiperda to the echolocation hunting sequence of Molossus molossus, a two-tone bat. We also manipulated the bat calls to control for the frequency shifts by lowering the frequency band of the search and approach calls. The firing response of the A₁ receptor cell significantly decreases with the shift to higher frequencies during the search and approach phases of the hunting sequence of M. molossus; this could be explained by the receptor's threshold curve. The frequency dependence of the decrease in the receptor's response is supported by the results attained with the manipulated sequence: search and approach calls with the same minimum frequency are detected by the moth at the same threshold intensity. The two-tone bat M. molossus shows a call frequency alternation behavior that may enable it to overcome moth audition even in the mid-frequency range (i.e. 20-50 kHz) where moths hear best.