High duty cycle moth sounds jam bat echolocation: bats counter with compensatory changes in buzz duration.

Tiger moth species vary greatly in the number of clicks they produce and the resultant duty cycle. Signals with higher duty cycles are expected to more effectively interfere with bat sonar. However, little is known about the minimum duty cycle of tiger moth signals for sonar jamming. Is there a threshold that allows us to classify moths as acoustically aposematic versus sonar jammers based on their duty cycles? We performed playback experiments with three wild-caught adult male bats, Eptesicus fuscus. Bat attacks on tethered moths were challenged using acoustic signals of Bertholdia trigona with modified duty cycles ranging from 0 to 46%. We did not find evidence for a duty cycle threshold; rather, the ability to jam the bat's sonar was a continuous function of duty cycle consistent with a steady increase in the number of clicks arriving during a critical signal processing time window just prior to the arrival of an echo. The proportion of successful captures significantly decreased as the moth duty cycle increased. Our findings suggest that moths cannot be unambiguously classified as acoustically aposematic or sonar jammers based solely on duty cycle. Bats appear to compensate for sonar jamming by lengthening the duration of their terminal buzz and they are more successful in capturing moths when they do so. In contrast to previous findings for bats performing difficult spatial tasks, the number of sonar sound groups decreased in response to high duty cycles and did not affect capture success.

Spontaneous motor-behavior abnormalities in two Drosophila models of neurodevelopmental disorders.

Mutations in hundreds of genes cause neurodevelopmental disorders with abnormal motor behavior alongside cognitive deficits. Boys with fragile X syndrome (FXS), a leading monogenic cause of intellectual disability, often display repetitive behaviors, a core feature of autism. By direct observation and manual analysis, we characterized spontaneous-motor-behavior phenotypes of Drosophila dfmr1 mutants, an established model for FXS. We recorded individual 1-day-old adult flies, with mature nervous systems and prior to the onset of aging, in small arenas. We scored behavior using open-source video-annotation software to generate continuous activity timelines, which were represented graphically and quantitatively. Young dfmr1 mutants spent excessive time grooming, with increased bout number and duration; both were rescued by transgenic wild-type dfmr1+. By two grooming-pattern measures, dfmr1-mutant flies showed elevated repetitions consistent with perseveration, which is common in FXS. In addition, the mutant flies display a preference for grooming posterior body structures, and an increased rate of grooming transitions from one site to another. We raise the possibility that courtship and circadian rhythm defects, previously reported for dfmr1 mutants, are complicated by excessive grooming. We also observed significantly increased grooming in CASK mutants, despite their dramatically decreased walking phenotype. The mutant flies, a model for human CASK-related neurodevelopmental disorders, displayed consistently elevated grooming indices throughout the assay, but transient locomotory activation immediately after placement in the arena. Based on published data identifying FMRP-target transcripts and functional analyses of mutations causing human genetic neurodevelopmental disorders, we propose the following proteins as candidate mediators of excessive repetitive behaviors in FXS: CaMKIIα, NMDA receptor subunits 2A and 2B, NLGN3, and SHANK3. Together, these fly-mutant phenotypes and mechanistic insights provide starting points for drug discovery to identify compounds that reduce dysfunctional repetitive behaviors.

Characteristics of tiger moth (Erebidae: Arctiinae) anti-bat sounds can be predicted from tymbal morphology.

BACKGROUND: Acoustic signals are used by many animals to transmit information. Variation in the acoustic characteristics of these signals often covaries with morphology and can relay information about an individual's fitness, sex, species, and/or other characteristics important for both mating and defense. Tiger moths (Lepidoptera: Erebidae: Arctiinae) use modified cuticular plates called "tymbal organs" to produce ultrasonic clicks which can aposematically signal their toxicity, mimic the signals of other species, or, in some cases, disrupt bat echolocation. The morphology of the tymbal organs and the sounds they produce vary greatly between species, but it is unclear how the variation in morphology gives rise to the variation in acoustic characteristics. This is the first study to determine how the morphological features of tymbals can predict the acoustic characteristics of the signals they produce. RESULTS: We show that the number of striations on the tymbal surface (historically known as "microtymbals") and, to a lesser extent, the ratio of the projected surface area of the tymbal to that of the thorax have a strong, positive correlation with the number of clicks a moth produces per unit time. We also found that some clades have significantly different regression coefficients, and thus the relationship between microtymbals and click rate is also dependent on the shared ancestry of different species. CONCLUSIONS: Our predictive model allows the click rates of moths to be estimated using preserved material (e.g., from museums) in cases where live specimens are unavailable. This has the potential to greatly accelerate our understanding of the distribution of sound production and acoustic anti-bat strategies employed by tiger moths. Such knowledge will generate new insights into the evolutionary history of tiger moth anti-predator defenses on a global scale.

The jamming avoidance response in echolocating bats.

Bats face many sources of acoustic interference in their natural environments, including other bats and potential prey items that affect their ability to interpret the returning echoes of their biosonar signals. To be able to navigate and forage successfully, bats must be able to counteract this interference and one of the ways they achieve this is by altering the various parameters of their echolocation. We describe these changes in signal design within the context of a modified definition of the jamming avoidance response originally applied to the signal changes of weakly electric fish. Both of these groups use active sensory systems that exhibit similarities in function but we take this opportunity to highlight major differences each groups' response to signal interference. These discrepancies form the basis of our need for an expanded description of the jamming avoidance response in echolocating bats.

Active acoustic interference elicits echolocation changes in heterospecific bats.

Echolocating bats often forage in the presence of both conspecific and heterospecific individuals, which have the potential to produce acoustic interference. Recent studies have shown that at least one bat species, the Brazilian free-tailed bat (Tadarida brasiliensis), produces specialized social signals that disrupt the sonar of conspecific competitors. We herein discuss the differences between passive and active jamming signals and test whether heterospecific jamming occurs in species overlapping spatiotemporally, as well as whether such interference elicits a jamming avoidance response. We compare the capture rates of tethered moths and the echolocation parameters of big brown bats (Eptesicus fuscus) challenged with the playback of the jamming signal normally produced by Brazilian free-tailed bats and playback of deconstructed versions of this signal. There were no differences in the capture rates of targets with and without the jamming signal, although significant changes in both spectral and temporal features of the bats' echolocation were observed. These changes are consistent with improvements of the signal-to-noise ratio in the presence of acoustic interference. Accordingly, we propose to expand the traditional definition of the jamming avoidance response, stating that echolocation changes in response to interference should decrease similarity between the two signals, to include any change that increases the ability to separate returning echoes from active jamming stimuli originating from conspecific and heterospecific organisms. Flexibility in echolocation is an important characteristic for overcoming various forms of acoustic interference and may serve a purpose in interspecific interactions as well as intraspecific ones.

How moths escape bats: predicting outcomes of predator-prey interactions.

What determines whether fleeing prey escape from attacking predators? To answer this question, biologists have developed mathematical models that incorporate attack geometries, pursuit and escape trajectories, and kinematics of predator and prey. These models have rarely been tested using data from actual predator-prey encounters. To address this problem, we recorded multi-camera infrared videography of bat-insect interactions in a large outdoor enclosure. We documented 235 attacks by four Myotis volans bats on a variety of moths. Bat and moth flight trajectories from 50 high-quality attacks were reconstructed in 3-D. Despite having higher maximum velocity, deceleration and overall turning ability, bats only captured evasive prey in 69 of 184 attacks (37.5%); bats captured nearly all moths not evading attack (50 of 51; 98%). Logistic regression indicated that prey radial acceleration and escape angle were the most important predictors of escape success (44 of 50 attacks correctly classified; 88%). We found partial support for the turning gambit mathematical model; however, it underestimated the escape threshold by 25% of prey velocity and did not account for prey escape angle. Whereas most prey escaping strikes flee away from predators, moths typically escaped chasing bats by turning with high radial acceleration toward 'safety zones' that flank the predator. This strategy may be widespread in prey engaged in chases. Based on these findings, we developed a novel geometrical model of predation. We discuss implications of this model for the co-evolution of predator and prey kinematics and pursuit and escape strategies.

Acoustic Aposematism and Evasive Action in Select Chemically Defended Arctiine (Lepidoptera: Erebidae) Species: Nonchalant or Not?

Tiger moths (Erebidae: Arctiinae) have experienced intense selective pressure from echolocating, insectivorous bats for over 65 million years. One outcome has been the evolution of acoustic signals that advertise the presence of toxins sequestered from the moths' larval host plants, i.e. acoustic aposematism. Little is known about the effectiveness of tiger moth anti-bat sounds in their natural environments. We used multiple infrared cameras to reconstruct bat-moth interactions in three-dimensional (3-D) space to examine how functional sound-producing organs called tymbals affect predation of two chemically defended tiger moth species: Pygarctia roseicapitis (Arctiini) and Cisthene martini (Lithosiini). P. roseicapitis and C. martini with intact tymbals were 1.8 and 1.6 times less likely to be captured by bats relative to those rendered silent. 3-D flight path and acoustic analyses indicated that bats actively avoided capturing sound-producing moths. Clicking behavior differed between the two tiger moth species, with P. roseicapitis responding in an earlier phase of bat attack. Evasive flight behavior in response to bat attacks was markedly different between the two tiger moth species. P. roseicapitis frequently paired evasive dives with aposematic sound production. C. martini were considerably more nonchalant and employed evasion in fewer interactions. Our results show that acoustic aposematism is effective at deterring bat predation in a natural context and that this strategy is likely to be the ancestral function of tymbal organs within the Arctiinae.

Bats jamming bats: food competition through sonar interference.

Communication signals are susceptible to interference ("jamming") from conspecifics and other sources. Many active sensing animals, including bats and electric fish, alter the frequency of their emissions to avoid inadvertent jamming from conspecifics. We demonstrated that echolocating bats adaptively jam conspecifics during competitions for food. Three-dimensional flight path reconstructions and audio-video field recordings of foraging bats (Tadarida brasiliensis) revealed extended interactions in which bats emitted sinusoidal frequency-modulated ultrasonic signals that interfered with the echolocation of conspecifics attacking insect prey. Playbacks of the jamming call, but not of control sounds, caused bats to miss insect targets. This study demonstrates intraspecific food competition through active disruption of a competitor's sensing during food acquisition.

Optimal predator risk assessment by the sonar-jamming arctiine moth Bertholdia trigona.

Nearly all animals face a tradeoff between seeking food and mates and avoiding predation. Optimal escape theory holds that an animal confronted with a predator should only flee when benefits of flight (increased survival) outweigh the costs (energetic costs, lost foraging time, etc.). We propose a model for prey risk assessment based on the predator's stage of attack. Risk level should increase rapidly from when the predator detects the prey to when it commits to the attack. We tested this hypothesis using a predator--the echolocating bat--whose active biosonar reveals its stage of attack. We used a prey defense--clicking used for sonar jamming by the tiger moth Bertholdia trigona--that can be readily studied in the field and laboratory and is enacted simultaneously with evasive flight. We predicted that prey employ defenses soon after being detected and targeted, and that prey defensive thresholds discriminate between legitimate predatory threats and false threats where a nearby prey is attacked. Laboratory and field experiments using playbacks of ultrasound signals and naturally behaving bats, respectively, confirmed our predictions. Moths clicked soon after bats detected and targeted them. Also, B. trigona clicking thresholds closely matched predicted optimal thresholds for discriminating legitimate and false predator threats for bats using search and approach phase echolocation--the period when bats are searching for and assessing prey. To our knowledge, this is the first quantitative study to correlate the sensory stimuli that trigger defensive behaviors with measurements of signals provided by predators during natural attacks in the field. We propose theoretical models for explaining prey risk assessment depending on the availability of cues that reveal a predator's stage of attack.

Sonar jamming in the field: effectiveness and behavior of a unique prey defense.

Bats and insects provide a model system for integrating our understanding of predator-prey ecology, animal behavior and neurophysiology. Previous field studies of bat-insect interactions have been limited by the technological challenges involved with studying nocturnal, volant animals that use ultrasound and engage in battles that frequently last a fraction of a second. We overcame these challenges using a robust field methodology that included multiple infrared cameras calibrated for three-dimensional reconstruction of bat and moth flight trajectories and four ultrasonic microphones that provided a spatial component to audio recordings. Our objectives were to document bat-moth interactions in a natural setting and to test the effectiveness of a unique prey defense - sonar jamming. We tested the effect of sonar jamming by comparing the results of interactions between bats and Grote's tiger moth, Bertholdia trigona, with their sound-producing organs either intact or ablated. Jamming was highly effective, with bats capturing more than 10 times as many silenced moths as clicking moths. Moths frequently combined their acoustic defense with two separate evasive maneuvers: flying away from the bat and diving. Diving decreased bat capture success for both clicking and silenced moths, while flying away did not. The diving showed a strong directional component, a first for insect defensive maneuvers. We discuss the timing of B. trigona defensive maneuvers - which differs from that of other moths - in the context of moth auditory neuroethology. Studying bat-insect interactions in their natural environment provides valuable information that complements work conducted in more controlled settings.

Sound strategies: the 65-million-year-old battle between bats and insects.

The intimate details regarding the coevolution of bats and moths have been elucidated over the past 50 years. The bat-moth story began with the evolution of bat sonar, an exquisite ultrasonic system for tracking prey through the night sky. Moths countered with ears tuned to the high frequencies of bat echolocation and with evasive action through directed turns, loops, spirals, drops, and power dives. Some bat species responded by moving the frequency and intensity of their echolocation cries away from the peak sensitivity of moth ears, and the arms race was on. Tiger moths countered by producing anti-bat sounds. Do the sounds advertise moth toxicity, similar to the bright coloration of butterflies; do they startle the bat, giving the moth a momentary advantage in their aerobatic battle; or do they jam the sonar of the bat? The answer is yes. They do all and more in different situations and in different species. Any insect that flies at night must deal with bat predation. Beetles, mantids, true crickets, mole crickets, katydids, green lacewings, and locusts have anti-bat strategies, and we have just scratched the surface. In an exciting new twist, researchers are taking the technologies developed in the laboratory back into the field, where they are poised to appreciate the full richness of this remarkable predator-prey interaction.

How do tiger moths jam bat sonar?

The tiger moth Bertholdia trigona is the only animal in nature known to defend itself by jamming the sonar of its predators - bats. In this study we analyzed the three-dimensional flight paths and echolocation behavior of big brown bats (Eptesicus fuscus) attacking B. trigona in a flight room over seven consecutive nights to determine the acoustic mechanism of the sonar-jamming defense. Three mechanisms have been proposed: (1) the phantom echo hypothesis, which states that bats misinterpret moth clicks as echoes; (2) the ranging interference hypothesis, which states that moth clicks degrade the bats' precision in determining target distance; and (3) the masking hypothesis, which states that moth clicks mask the moth echoes entirely, making the moth temporarily invisible. On nights one and two of the experiment, the bats appeared startled by the clicks; however, on nights three through seven, the bats frequently missed their prey by a distance predicted by the ranging interference hypothesis (∼15-20 cm). Three-dimensional simulations show that bats did not avoid phantom targets, and the bats' ability to track clicking prey contradicts the predictions of the masking hypothesis. The moth clicks also forced the bats to reverse their stereotyped pattern of echolocation emissions during attack, even while bats continued pursuit of the moths. This likely further hinders the bats' ability to track prey. These results have implications for the evolution of sonar jamming in tiger moths, and we suggest evolutionary pathways by which sonar jamming may have evolved from other tiger moth defense mechanisms.

Tiger moth jams bat sonar.

In response to sonar-guided attacking bats, some tiger moths make ultrasonic clicks of their own. The lepidopteran sounds have previously been shown to alert bats to some moths' toxic chemistry and also to startle bats unaccustomed to sonic prey. The moth sounds could also interfere with, or "jam," bat sonar, but evidence for such jamming has been inconclusive. Using ultrasonic recording and high-speed infrared videography of bat-moth interactions, we show that the palatable tiger moth Bertholdia trigona defends against attacking big brown bats (Eptesicus fuscus) using ultrasonic clicks that jam bat sonar. Sonar jamming extends the defensive repertoire available to prey in the long-standing evolutionary arms race between bats and insects.

Naïve bats discriminate arctiid moth warning sounds but generalize their aposematic meaning.

Naïve red (Lasiurus borealis Müller) and big brown (Eptesicus fuscus Beauvois) bats quickly learn to avoid noxious sound-producing tiger moths. After this experience with a model tiger moth, bats generalize the meaning of these prey-generated sounds to a second tiger moth species producing a different call. Here we describe the three-dimensional kinematic and bioacoustic details of this behaviour, first, as naïve bats learn to deal with an unpalatable model tiger moth and subsequently, as they avoid acoustic mimics. The tiger moths' first clicks influenced the bats' echolocation behaviour and the percentage of interactions that included terminal buzzes was associated with capture and investigatory behaviour. When the mimic was introduced, the bats decreased both their minimum distance to the tiger moth and the time at which they broke off their attack compared with their exposure to the model on the night before. These kinematic signatures closely match the bats' behaviour on their first night of experience with the model. Minimum distances and time of pursuit cessation increased again by the last night of the mimic's presentation. These kinematic and bioacoustic results show that although naïve bats generalize the meaning of aposematic tiger moth calls, they discriminate the prey-generated signals as different and investigate. Extrapolating to experienced bats, these results suggest that acoustic predators probably exert potent and fine-scaled selective forces on acoustic mimicry complexes.

Has vertebrate chemesthesis been a selective agent in the evolution of arthropod chemical defenses?

Arthropods use a variety of chemical substances to repel potential predators, but how did they arrive at the suite of chemicals that they use? One way to explore this question is to map chemically defended arthropod species in a multidimensional "compound" space. Clustering within this space indicates species that share similar combinations of chemical compounds and can reflect a phylogenetic signal, common biochemical pathways, or both. More important for this study, clustering can help to identify allomone targets. We herein compare common arthropod allomones with known vertebrate trigeminal irritants. We argue that the degree of overlap between these two groups of compounds indicates that chemesthesis was an important determining factor in the evolution of many arthropod allomones. The multidimensional scaling methods used may also allow the identification of new irritant receptors.

Morphogenetic effects of alkaloidal metabolites on the development of the coremata in the salt marsh moth, Estigmene acrea (Dru.) (Lepidoptera: Arctiidae).

Pyrrolizidine alkaloids (PAs) play a fundamental role in the sexual biology of the salt marsh moth Estigmene acrea. They are precursors for the male courtship pheromone hydroxydanaidal and they stimulate the growth and development of male pheromone-disseminating organs called coremata. Yet larval Estigmene are polyphagous and feed only sporadically on PA-containing plants and those they utilize contain different classes of PAs. The various PAs ingested are hydrolyzed to the common necine metabolite retronecine and re-esterified to insect-specific alkaloids from which the male pheromone hydroxydanaidal is synthesized. Given this complex metabolic pathway, we investigated the role of retronecine and the insect-specific alkaloids that stem from it as morphogens stimulating corematal growth. Retronecine fed to terminal instar larvae in a standard caterpillar diet stimulated corematal growth. It also stimulated corematal growth when it was injected into the hemolymph of larvae. These results indicate that this common PA metabolite, and/or the insect specific alkaloids produced from it, function as corematal morphogens. The parental forms (alkaloids ingested from the plant) are not strictly necessary for corematal growth. Stimulation of the PA receptors on the galea and ingestion process are also not critical to corematal development. Since the insect-specific alkaloids are the direct precursors for the male courtship pheromone, it is argued that their level is the best indicator of the ultimate pheromone titer and would provide the most accurate developmental signal. The effects of alkaloidal metabolites as morphogens in E. acrea are compared to those for the South Asian arctiines Creatonotus gangis and C. transiens in which the developmental role of PAs was first discovered.

Acoustic mimicry in a predator-prey interaction.

Mimicry of visual warning signals is one of the keystone concepts in evolutionary biology and has received substantial research attention. By comparison, acoustic mimicry has never been rigorously tested. Visualizing bat-moth interactions with high-speed, infrared videography, we provide empirical evidence for acoustic mimicry in the ultrasonic warning sounds that tiger moths produce in response to echolocating bats. Two species of sound-producing tiger moths were offered successively to naïve, free-flying red and big brown bats. Noctuid and pyralid moth controls were also offered each night. All bats quickly learned to avoid the noxious tiger moths first offered to them, associating the warning sounds with bad taste. They then avoided the second sound-producing species regardless of whether it was chemically protected or not, verifying both Müllerian and Batesian mimicry in the acoustic modality. A subset of the red bats subsequently discovered the palatability of the Batesian mimic, demonstrating the powerful selective force these predators exert on mimetic resemblance. Given these results and the widespread presence of tiger moth species and other sound-producing insects that respond with ultrasonic clicks to bat attack, acoustic mimicry complexes are likely common components of the acoustic landscape.

If you've got it, flaunt it: ingested alkaloids affect corematal display behavior in the salt marsh moth, Estigmene acrea.

Plant-derived pyrrolizidine alkaloids play an important role in the biology of the salt marsh moth, Estigmene acrea (Lepidoptera: Arctiidae). They are phagostimulants for larvae and they stimulate the growth and development of adult male androconial organs called coremata. In this study, we have shown that the pyrrolizidine alkaloid monocrotaline N-oxide (MNO) fed to larvae also affects the courtship behavior of adult males. Males fed a diet containing MNO display their coremata while males fed on the same diet without alkaloid rarely display. This explains why it has been difficult to replicate field observations of the "lekking" behavior of this species in the laboratory where animals are frequently raised on commercially available diets devoid of alkaloids. Corematal inflation was observed in isolated males and in laboratory leks. The effect of larvae feeding on pyrrolizidine alkaloid on the reproductive behavior of adults suggests that this substance may modify the development of the moth's nervous system and contribute to their unusual dual mating strategies. MNO was also shown to be an adequate precursor for the production of the courtship pheromone hydroxydanaidal.

Sound strategy: acoustic aposematism in the bat-tiger moth arms race.

The night sky is the venue for an ancient arms race. Insectivorous bats with their ultrasonic sonar exert an enormous selective pressure on nocturnal insects. In response insects have evolved the ability to hear bat cries, to evade their hunting maneuvers, and some, the tiger moths (Arctiidae), to utter an ultrasonic reply. We here determine what it is that tiger moths "say" to bats. We chose four species of arctiid moths, Cycnia tenera, Euchaetes egle, Utetheisa ornatrix, and Apantesis nais, that naturally differ in their levels of unpalatability and their ability to produce sound. Moths were tethered and offered to free-flying naive big brown bats, Eptesicus fuscus. The ability of the bats to capture each species was compared to their ability to capture noctuid, geometrid, and wax moth controls over a learning period of 7 days. We repeated the experiment using the single arctiid species E. egle that through diet manipulation and simple surgery could be rendered palatable or unpalatable and sound producing or mute. We again compared the capture rates of these categories of E. egle to control moths. Using both novel learning approaches we have found that the bats only respond to the sounds of arctiids when they are paired with defensive chemistry. The sounds are in essence a warning to the bats that the moth is unpalatable-an aposematic signal.

Interspecific differences in Aethia spp. auklet odorants and evidence for chemical defense against ectoparasites.

The true auklets (Genus Aethia) are small planktivorous seabirds of the Bering Sea and North Pacific. Two species, the crested and whiskered auklets produce volatile citrus-like odorants. We here show that the whiskered auklet odorant is composed predominantly of two odd-numbered aldehydes (heptanal and nonanal) with no detectable unsaturated aldehydes. By comparison the crested auklet odorant is dominated by even-numbered aldehydes, both saturated and monounsaturated, ranging in size from 6 to 12 carbons. This is evidence of species-specific acquisition or biosynthetic pathways. We clarify the chemistry of the crested auklet odorant. We cite evidence that the C-12:1 aldehyde in crested auklets is actually two isomers, (Z)-4-dodecenal and (Z)-6-dodecenal. We also report on experimental evidence that aldehyde constituents kill and repel ectoparasites. Efficacy of the aldehydes may increase when they are combined in a mixture. The repellency of the mixture increases with chemical concentration. This suggests that individuals with higher chemical production are likely to repel ectoparasites more effectively.