Mechanics of a 'simple' ear: tympanal vibrations in noctuid moths.

Anatomically, the ears of moths are considered to be among the simplest ears found in animals. Microscanning laser vibrometry was used to examine the surface vibrations of the entire tympanal region of the ears of the noctuid moths Agrotis exclamationis, Noctua pronuba, Xestia c-nigrum and Xestia triangulum. During stimulation with ultrasound at intensities known to activate receptor neurones, the tympanum vibrates with maximum deflection amplitudes at the location where the receptor cells attach. In the reportedly heterogeneous tympana of noctuid moths, this attachment site is an opaque zone that is surrounded by a transparent, thinner cuticular region. In response to sound pressure, this region moves relatively little compared with the opaque zone. Thus, the deflections of the moth tympanic membrane are not those of a simple circular drum. The acoustic sensitivity of the ear of N. pronuba, as measured on the attachment site, is 100+/-14 nm Pa(-1) (N=10), corresponding to tympanal motion of a mere 200 pm at sound pressure levels near the neural threshold.

Extinction of the acoustic startle response in moths endemic to a bat-free habitat.

Most moths use ears solely to detect the echolocation calls of hunting, insectivorous bats and evoke evasive flight manoeuvres. This singularity of purpose predicts that this sensoribehavioural network will regress if the selective force that originally maintained it is removed. We tested this with noctuid moths from the islands of Tahiti and Moorea, sites where bats have never existed and where an earlier study demonstrated that the ears of endemic species resemble those of adventives although partially reduced in sensitivity. To determine if these moths still express the anti-bat defensive behaviour of acoustic startle response (ASR) we compared the nocturnal flight times of six endemic to six adventive species in the presence and absence of artificial bat echolocation sounds. Whereas all of the adventive species reduced their flight times when exposed to ultrasound, only one of the six endemic species did so. These differences were significant when tested using a phylogenetically based pairwise comparison and when comparing effect sizes. We conclude that the absence of bats in this habitat has caused the neural circuitry that normally controls the ASR behaviour in bat-exposed moths to become decoupled from the functionally vestigial ears of endemic Tahitian moths.

Auditory sensitivity of Hawaiian moths (Lepidoptera: Noctuidae) and selective predation by the Hawaiian hoary bat (Chiroptera: Lasiurus cinereus semotus).

The islands of Hawai'i offer a unique opportunity for studying the auditory ecology of moths and bats since this habitat has a single species of bat, the Hawaiian hoary bat (Lasiurus cinereus semotus), which exerts the entire predatory selection pressure on the ears of sympatric moths. I compared the moth wings discarded by foraging bats with the number of surviving moths on the island of Kaua'i and concluded that the endemic noctuid Haliophyle euclidias is more heavily preyed upon than similar-sized endemic (e.g. Agrotis diplosticta) and adventive (Agrotis ipsilon and Pseudaletia unipuncta) species. Electrophysiological examinations indicated that, compared with species less preyed upon, H. euclidias has lower auditory sensitivities to the bat's social and echolocation calls, which will result in shorter detection distances of the bat. The poor ears of H. euclidias suggest that this moth coevolved with the bat using non-auditory defences that resulted in auditory degeneration. This moth now suffers higher predation because it is drawn away from its normal habitat by the man-made lights that are exploited by the bat.

Sound production and hearing in the blue cracker butterfly Hamadryas feronia (Lepidoptera, nymphalidae) from Venezuela.

Certain species of Hamadryas butterflies are known to use sounds during interactions with conspecifics. We have observed the behaviour associated with sound production and report on the acoustic characteristics of these sounds and on the anatomy and physiology of the hearing organ in one species, Hamadryas feronia, from Venezuela. Our observations confirm previous reports that males of this species will take flight from their tree perch when they detect a passing conspecific (male or female) and, during the chase, produce clicking sounds. Our analyses of both hand-held males and those flying in the field show that the sounds are short (approximately 0.5 s) trains of intense (approximately 80-100 dB SPL at 10 cm) and brief (2-3 ms) double-component clicks, exhibiting a broad frequency spectrum with a peak energy around 13-15 kHz. Our preliminary results on the mechanism of sound production showed that males can produce clicks using only one wing, thus contradicting a previous hypothesis that it is a percussive mechanism. The organ of hearing is believed to be Vogel's organ, which is located at the base of the forewing subcostal and cubital veins. Vogel's organ consists of a thinned region of exoskeleton (the tympanum) bordered by a rigid chitinous ring; associated with its inner surface are three chordotonal sensory organs and enlarged tracheae. The largest chordotonal organ attaches to a sclerite positioned near the center of the eardrum and possesses more than 110 scolopidial units. The two smaller organs attach to the perimeter of the membrane. Extracellular recordings from the nerve branch innervating the largest chordotonal organ confirm auditory sensitivity with a threshold of 68 dB SPL at the best frequency of 1.75 kHz. Hence, the clicks with peak energy around 14 kHz are acoustically mismatched to the best frequencies of the ear. However, the clicks are broad-banded and even at 1-2 kHz, far from the peak frequency, the energy is sufficient such that the butterflies can easily hear each other at the close distances at which they interact (less than 30 cm). In H. feronia, Vogel's organ meets the anatomical and functional criteria for being recognized as a typical insect tympanal ear.

Day-flying butterflies remain day-flying in a Polynesian, bat-free habitat.

To test the theory that insectivorous bats have selected for diurnality in earless butterflies I compared the nocturnal flight patterns of three species of nymphalid butterflies on the bat-free Pacific island of Moorea with those of three nymphalids in the bat-inhabited habitat of Queensland, Australia. Nocturnal flight, measured as the ratio of deep night (1 h following sunset to 1 h preceding sunrise) to twilight night (1 h before sunset to 30 min after sunrise) activity did not differ significantly between the two locations, nor did the percentage of individuals active and I conclude that living in a bat-released habitat has not produced nocturnal flight in these insects. This result is surprising considering the potential advantages of escaping diurnally active predators and suggests that physiological adaptations (e.g. thermoregulation and/or vision) currently constrain these insects to diurnal flight. Since taxonomic records suggest that gene flow does not exist with bat-exposed conspecifics, I suggest that insufficient time has elapsed since these species migrated to Moorea to have resulted in major phenotypic changes such as diel flight preferences.

Bat-deafness in day-flying moths (Lepidoptera, Notodontidae, Dioptinae).

Assuming that bat-detection is the primary function of moth ears, the ears of moths that are no longer exposed to bats should be deaf to echolocation call frequencies. To test this, we compared the auditory threshold curves of 7 species of Venezuelan day-flying moths (Notodontidae: Dioptinae) to those of 12 sympatric species of nocturnal moths (Notodontidae: Dudusinae, Noctuidae and Arctiidae). Whereas 2 dioptines (Josia turgida, Zunacetha annulata) revealed normal ears, 2 (J. radians, J. gopala) had reduced hearing at bat-specific frequencies (20-80 kHz) and the remaining 3 (Thirmida discinota, Polypoetes circumfumata and Xenorma cytheris) revealed pronounced to complete levels of high-frequency deafness. Although the bat-deaf ears of dioptines could function in other purposes (e.g., social communication), the poor sensitivities of these species even at their best frequencies suggest that these moths represent a state of advanced auditory degeneration brought about by their diurnal life history. The phylogeny of the Notodontidae further suggests that this deafness is a derived (apomorphic) condition and not a retention of a primitive (pleisiomorphic), insensitive state.

Neurometamorphosis of the ear in the gypsy moth, Lymantria dispar, and its homologue in the earless forest tent caterpillar moth, Malacosoma disstria.

The adult gypsy moth, Lymantria dispar (Lymantriidae: Noctuoidea) has a pair of metathoracic tympanic ears that each contain a two-celled auditory chordotonal organ (CO). The earless forest tent caterpillar moth, Malacosoma disstria (Lasiocampidae: Bombycoidea), has a homologous pair of three-celled, nonauditory hindwing COs in their place. The purpose of our study was to determine whether the adult CO in both species arises from a preexisting larval organ or if it develops as a novel structure during metamorphosis. We describe the larval metathoracic nervous system of L. dispar and M. disstria, and identify a three-celled chordotonal organ in the anatomically homologous site as the adult CO. If the larval CO is severed from the homologue of the adult auditory nerve (IIIN1b1) in L. dispar prior to metamorphosis, the adult develops an ear lacking an auditory organ. Axonal backfills of the larval IIIN1b1 nerve in both species reveal three chordotonal sensory neurons and one nonchordotonal multipolar cell. The axons of these cells project into tracts of the central nervous system putatively homologous with those of the auditory pathway in adult L. dispar. Following metamorphosis, M. disstria moths retain all four cells (three CO and one multipolar) while L. dispar adults possess two cells that service the auditory CO and one nonauditory, multipolar cell. We conclude that the larval IIIN1b1 CO is the precursor of both the auditory organ in L. dispar and the putative proprioceptor CO in M. disstria and represents the premetamorphic condition of these insects. The implications of our results in understanding the evolution of the ear in the Lepidoptera and insects in general are discussed.

The closed-loop nature of the tymbal response in the dogbane tiger moth, Cycnia tenera (Lepidoptera, Arctiidae).

The dogbane tiger moth, Cycnia tenera, emits ultrasonic sounds by rhythmically buckling a pair of tymbals when stimulated by pulsed sounds resembling bat echolocation. We monitored the central pattern generator governing this response by recording the motor output of the tymbal branch of the metathoracic leg nerve. The rhythm of the tymbal motor pattern can be altered midway (500 m/sec from its initiation) by changing the period and, to a lesser degree, the intensity of the stimulus. The tymbal response of C. tenera is therefore closed-looped to stimulus pulse period and intensity. Our results suggest that C. tenera relies less on the changes in an attacking bat's echolocation intensity when responding with this behaviour because this acoustic parameter may be a more unreliable indicator of the proximity of the bat than its echolocation call period.

Jamming bat echolocation: the dogbane tiger moth Cycnia tenera times its clicks to the terminal attack calls of the big brown bat Eptesicus fuscus.

Certain tiger moths emit high-frequency clicks to an attacking bat, causing it to break off its pursuit. The sounds may either orient the bat by providing it with information that it uses to make an attack decision (aposematism) or they may disorient the bat by interrupting the normal flow of echo information required to complete a successful capture (startle, jamming). At what point during a bat's attack does an arctiid emit its clicks? If the sounds are aposematic, the moth should emit them early in the attack echolocation sequence in order to allow the bat time to understand their meaning. If, however, the sounds disrupt the bat's echo-processing behaviour, one would expect them to be emitted later in the attack to maximize their confusion effects. To test this, we exposed dogbane tiger moths (Cycnia tenera) to a recording of the echolocation sequence emitted by a big brown bat (Eptesicus fuscus) as it attacked a stationary target. Our results demonstrate that, at normal echolocation intensities, C. tenera does not respond to approach calls but waits until the terminal phase of the attack before emitting its clicks. This timing is evident whether the moth is stationary or flying and is largely independent of the intensity of the echolocation calls. These results support the hypothesis of a jamming effect (e.g. 'phantom echoes') and suggest that, to determine experimentally the effects of arctiid clicks on bats, it is important that the bats be tested under conditions that simulate the natural context in which this defence operates.

The gleaning attacks of the northern long-eared bat, Myotis septentrionalis, are relatively inaudible to moths.

This study empirically tests the prediction that the echolocation calls of gleaning insectivorous bats (short duration, high frequency, low intensity) are acoustically mismatched to the ears of noctuid moths and are less detectable than those of aerially hawking bats. We recorded auditory receptor cell action potentials elicited in underwing moths (Catocala spp.) by echolocation calls emitted during gleaning attacks by Myotis septentrionalis (the northern long-eared bat) and during flights by the aerial hawker Myotis lucifugus (the little brown bat). The moth ear responds inconsistently and with fewer action potentials to the echolocation calls emitted by the gleaner, a situation that worsened when the moth's ear was covered by its wing (mimicking a moth resting on a surface). Calls emitted by the aerial-hawking bat elicited a significantly stronger spiking response from the moth ear. Moths with their ears covered by their wings maintained their relative hearing sensitivity at their best frequency range, the range used by most aerial insectivorous bats, but showed a pronounced deafness in the frequency range typically employed by gleaning bats. Our results (1) support the prediction that the echolocation calls of gleaners are acoustically inconspicuous to the ears of moths (and presumably other nocturnal tympanate insects), leaving the moths particularly vulnerable to predation, and (2) suggest that gleaners gain a foraging advantage against eared prey.

The evolutionary biology of insect hearing.

Few areas of science have experienced such a blending of laboratory and field perspectives as the study of hearing. The disciplines of sensory ecology and neuroethology interpret the morphology and physiology of ears in the adaptive context in which this sense organ functions. Insects, with their enormous diversity, are valuable candidates for the study of how tympanal ears have evolved and how they operate today in different habitats.

The mechanoreceptive origin of insect tympanal organs: a comparative study of similar nerves in tympanate and atympanate moths.

A chordotonal organ occurring in the posterior metathorax of an atympanate moth, Actias luna (L.) (Bombycoidea: Saturniidae), appears to be homologous to the tympanal organ of the noctuoid moth. The peripheral anatomy of the metathoracic nerve branch, IIIN1b1 was examined in Actias luna with cobalt-lysine and Janus Green B, and compared to its counterpart, IIIN1b (the tympanal branch), in Feltia heralis (Grt.) (Noctuoidea: Noctuidae). The peripheral projections of IIIN1b1 were found to be similar in both species, dividing into three branches, the second (IIIN1b1b) ending as a chordotonal organ. The atympanate organ possesses three sensory cell bodies and three scolopales, and is anchored peripherally via an attachment strand to the undifferentiated membranous region underlying the hindwing alula, which corresponds to the tympanal region of the noctuoid metathorax. Extracellular recordings of the IIIN1b1 nerve in Actias luna revealed a large spontaneously active unit which fired in a regular pattern (corresponding to the noctuoid B cell) and smaller units (corresponding to the noctuoid acoustic A cells) which responded phasically to low frequency sounds (2 kHz) played at high intensities (83-96 dB, SPL) and also responded phasically to raising and lowering movements of the hindwing. We suggest that the chordotonal organ in Actias luna represents the evolutionary prototype to the noctuoid tympanal organ, and that it acts as a proprioceptor monitoring hindwing movements. This system, in its simplicity (consisting of only a few neurons) could be a useful model for examining the changes to the nervous system (both central and peripheral) that accompanied the evolutionary development of insect tympanal organs.

Functional organization of the arctiid moth tymbal (insecta, lepidoptera).

The exoskeletal morphology, muscular organization, and innervation patterns of the tymbals of seven sound-producing species of tiger moths (Arctiidae) were compared with the undifferentiated episterna of two silent species. At least three muscles are involved in sound production: the tymbal muscle, pv2, and the accessory muscles, pvl and/or pv6. All of the tymbal muscles are innervated by the IIIN2a branch of the metathoracic leg nerve, which contains two axons larger than the others. Backfills of the tymbal branch of the IIIN2a reveal a medial sensory neuropil and a population of five ipsilateral motor neurons whose somata are clustered into three groups along the anterior edge of the metathoracic ganglion. The dendritic arborizations of the motor neurons extend to the ganglionic midline but are separate from one part of the auditory neuropil observed in other noctuoids. The study concludes that the arctiid tymbal reveals only minor modifications (e.g., cuticle thinning) of the episterna of silent moths and represents a primitive form of the tymbal compared to those of the Cicadidae.

Information processing at a central synapse suggests a noise filter in the auditory pathway of the noctuid moth.

1. The central projections of the A1 afferent were confirmed via intracellular recording and staining with Lucifer Yellow in the pterothoracic ganglion of the noctuid moths, Agrotis infusa and Apamea amputatrix (Fig. 1). Simultaneous recordings of the A1 afferent in the tympanal nerve (extracellularly) and in the pterothoracic ganglion (intracellularly) confirm the identity of the stained receptor as being the A1 cell. 2. The major postsynaptic arborizations of interneurone 501 in the pterothoracic ganglion were also demonstrated via intracellular recording and staining (Fig. 2). Simultaneous recordings of the A1 afferent (extracellularly) and neurone 501 (intracellularly) revealed that each A1 spike evokes a constant short latency EPSP in the interneurone (Fig. 2Bi). Neurone 501 receives only monaural input from the A1 afferent on its soma side as demonstrated by electrical stimulation of each afferent nerve (Fig. 2Bii). EPSPs evoked in neurone 501 by high frequency (100 Hz) electrical stimulation of the afferent nerve did not decrement (Fig. 2Biii). These data are consistent with a monosynaptic input to neurone 501 from the A1 afferent. 3. The response of neurone 501 to a sound stimulus presented at an intensity near the upper limit of its linear response range (30 ms, 16 kHz, 80 dB SPL) was a plateau-like depolarization, with tonic spiking activity which continued beyond the end of the tone. The instantaneous spike frequency of the response was as high as 800 Hz, and was maintained at above 600 Hz for the duration of the tone (Fig. 3). 4. The relationship between the instantaneous spike frequency in the A1 afferent and that recorded simultaneously in neurone 501 is linear over the entire range of A1 spike frequencies evoked by white noise sound stimuli (Fig. 4). Similarly, the relationship between instantaneous spike frequency in the A1 afferent and the mean depolarization evoked in neurone 501 is also linear for all A1 spike frequencies tested (Fig. 5). No summation of EPSPs occurred for A1 spike frequencies below 100 Hz.(ABSTRACT TRUNCATED AT 250 WORDS)