Acoustic camouflage increases with body size and changes with bat echolocation frequency range in a community of nocturnally active Lepidoptera.

Body size is an important trait in predator-prey dynamics as it is often linked to detection, as well as the success of capture or escape. Larger prey, for example, often runs higher risk of detection by their predators, which imposes stronger selection on their anti-predator traits compared to smaller prey. Nocturnal Lepidoptera (moths) vary strongly in body size, which has consequences for their predation risk, as bigger moths return stronger echoes for echolocating bats. To compensate for increased predation risk, larger moths are therefore expected to have improved anti-predator defences. Moths are covered by different types of scales, which for a few species are known to absorb ultrasound, thus providing acoustic camouflage. Here, we assessed whether moths differ in their acoustic camouflage in a size-dependent way by focusing on their body scales and the different frequency ranges used by bats. We used a sonar head to measure 3D echo scans of a total of 111 moth specimens across 58 species, from eight different families of Lepidoptera. We scanned all the specimens and related their echo-acoustic target strength to various body size measurements. Next, we removed the scales covering the thorax and abdomen and scanned a subset of specimens again to assess the sound absorptive properties of these scales. Comparing intact specimens with descaled specimens, we found almost all species to absorb ultrasound, reducing detection risk on average by 8%. Furthermore, the sound absorptive capacities of body scales increased with body size suggesting that larger species benefit more from acoustic camouflage. The size-dependent effect of camouflage was in particular pronounced for the higher frequencies (above 29 kHz), with moth species belonging to large-bodied families consequently demonstrating similar target strengths compared to species from small-bodied families. Finally, we found the families to differ in frequency range that provided the largest reduction in detection risk, which may be related to differences in predation pressure and predator communities of these families. In general, our findings have important implications for predator-prey interactions across eco-evolutionary timescales and may suggest that acoustic camouflage played a role in body size evolution of nocturnally active Lepidoptera.

An ultrasound-absorbing inflorescence zone enhances echo-acoustic contrast of bat-pollinated cactus flowers.

Flowering plants have evolved an extraordinary variety of signalling traits to attract their pollinators. Most flowers rely on visual and chemical signals, but some bat-pollinated plants have evolved passive acoustic floral signals. All known acoustic flower signals rely on the same principle of increased sonar reflectivity. Here, we describe a novel mechanism that relies on increased absorption of the area surrounding the flower. In a bat-pollinated cactus (Espostoa frutescens) we found a hairy inflorescence zone, a so-called cephalium. Flowers solely emerge out of this zone. We measured the echoes of cephalia, flowers and unspecialized column surfaces and recorded echolocation calls of approaching bats. We found that the cephalium acts as a strong ultrasound absorber, attenuating the sound by -14 dB. The absorption was highest around the echolocation call frequencies of approaching bats. Our results indicate that, instead of making flowers more reflective, plants can also evolve structures to attenuate the background echo, thereby enhancing the acoustic contrast with the target.

Adding colour-realistic video images to audio playbacks increases stimulus engagement but does not enhance vocal learning in zebra finches.

Bird song and human speech are learned early in life and for both cases engagement with live social tutors generally leads to better learning outcomes than passive audio-only exposure. Real-world tutor-tutee relations are normally not uni- but multimodal and observations suggest that visual cues related to sound production might enhance vocal learning. We tested this hypothesis by pairing appropriate, colour-realistic, high frame-rate videos of a singing adult male zebra finch tutor with song playbacks and presenting these stimuli to juvenile zebra finches (Taeniopygia guttata). Juveniles exposed to song playbacks combined with video presentation of a singing bird approached the stimulus more often and spent more time close to it than juveniles exposed to audio playback only or audio playback combined with pixelated and time-reversed videos. However, higher engagement with the realistic audio-visual stimuli was not predictive of better song learning. Thus, although multimodality increased stimulus engagement and biologically relevant video content was more salient than colour and movement equivalent videos, the higher engagement with the realistic audio-visual stimuli did not lead to enhanced vocal learning. Whether the lack of three-dimensionality of a video tutor and/or the lack of meaningful social interaction make them less suitable for facilitating song learning than audio-visual exposure to a live tutor remains to be tested.

Acoustic traits of bat-pollinated flowers compared to flowers of other pollination syndromes and their echo-based classification using convolutional neural networks.

Bat-pollinated flowers have to attract their pollinators in absence of light and therefore some species developed specialized echoic floral parts. These parts are usually concave shaped and act like acoustic retroreflectors making the flowers acoustically conspicuous to the bats. Acoustic plant specializations only have been described for two bat-pollinated species in the Neotropics and one other bat-dependent plant in South East Asia. However, it remains unclear whether other bat-pollinated plant species also show acoustic adaptations. Moreover, acoustic traits have never been compared between bat-pollinated flowers and flowers belonging to other pollination syndromes. To investigate acoustic traits of bat-pollinated flowers we recorded a dataset of 32320 flower echoes, collected from 168 individual flowers belonging to 12 different species. 6 of these species were pollinated by bats and 6 species were pollinated by insects or hummingbirds. We analyzed the spectral target strength of the flowers and trained a convolutional neural network (CNN) on the spectrograms of the flower echoes. We found that bat-pollinated flowers have a significantly higher echo target strength, independent of their size, and differ in their morphology, specifically in the lower variance of their morphological features. We found that a good classification accuracy by our CNN (up to 84%) can be achieved with only one echo/spectrogram to classify the 12 different plant species, both bat-pollinated and otherwise, with bat-pollinated flowers being easier to classify. The higher classification performance of bat-pollinated flowers can be explained by the lower variance of their morphology.

Bio-acoustic tracking and localization using heterogeneous, scalable microphone arrays.

Microphone arrays are an essential tool in the field of bioacoustics as they provide a non-intrusive way to study animal vocalizations and monitor their movement and behavior. Microphone arrays can be used for passive localization and tracking of sound sources while analyzing beamforming or spatial filtering of the emitted sound. Studying free roaming animals usually requires setting up equipment over large areas and attaching a tracking device to the animal which may alter their behavior. However, monitoring vocalizing animals through arrays of microphones, spatially distributed over their habitat has the advantage that unrestricted/unmanipulated animals can be observed. Important insights have been achieved through the use of microphone arrays, such as the convergent acoustic field of view in echolocating bats or context-dependent functions of avian duets. Here we show the development and application of large flexible microphone arrays that can be used to localize and track any vocalizing animal and study their bio-acoustic behavior. In a first experiment with hunting pallid bats the acoustic data acquired from a dense array with 64 microphones revealed details of the bats' echolocation beam in previously unseen resolution. We also demonstrate the flexibility of the proposed microphone array system in a second experiment, where we used a different array architecture allowing to simultaneously localize several species of vocalizing songbirds in a radius of 75 m. Our technology makes it possible to do longer measurement campaigns over larger areas studying changing habitats and providing new insights for habitat conservation. The flexible nature of the technology also makes it possible to create dense microphone arrays that can enhance our understanding in various fields of bioacoustics and can help to tackle the analytics of complex behaviors of vocalizing animals.

Activity Budget Comparisons Using Long-Term Observations of a Group of Bottlenose Dolphins (Tursiops truncatus) under Human Care: Implications for Animal Welfare.

Zoos and aquaria must provide optimal husbandry conditions and the highest welfare standards for their animals. How the welfare state of an animal or a group of animals can be precisely assessed is currently under debate, and new approaches are necessary to reliably evaluate changes in welfare. One particular measure that is easily applicable is behavioral observations. However, for dolphins and other cetaceans under human care, reliable behavior-based measures are rare. Using long-term observations of a group of bottlenose dolphins, we investigate how their activity budgets and different behaviors changed over time and are impacted by construction noise. Additionally, we investigate whether behavioral differences are also reflected in changes in the dolphins' performance during daily training sessions. Our results show that construction noise significantly alters the dolphins' behavior. Play behavior decreases during construction periods; most importantly, the frequency of fast swimming activities significantly increased, and at the same time, a decrease in training performance is found. Additionally, inter- and intraindividual behavioral differences are attributed to factors, such as age or weaning. Significant changes in a dolphin's activity budget can also pose potential welfare concerns. Thus, this study highlights the importance of regularly assessing and analyzing the behavior of dolphins under human care. Behavioral observations are essential welfare indicators and can-when complemented with other measures, such as assessment of training performance-provide zoo staff with important information about each individual's state of welfare.

Saliva and Blood Cortisol Measurement in Bottlenose Dolphins (Tursiops truncatus): Methodology, Application, and Limitations.

A central task of zoos and aquaria is the frequent and accurate assessment of their animals' welfare. Recently, important steps have been made, such as the introduction of animal welfare evaluation tools and welfare decision trees. To determine animal welfare, it is not only important to collect life history data, such as longevity and reproductive success, but also for experienced observers or caretakers to conduct behavioral observations on a regular basis to assess animals' emotional state. To physiologically validate welfare observations, glucocorticoid levels are usually assessed, as they are a common indicator of stress. While, for many animals, these levels can be easily determined via fecal or hair samples, for cetaceans, the levels are usually determined via blood samples. As blood samples cannot be taken very frequently and the process may cause stress to the animals (if the samples are not taken following medical training), other techniques, such as the measurement of health biomarkers (especially cortisol, which can be measured in saliva), have become the focus of cetacean stress research. However, there are two problems associated with saliva measurements in cetaceans: saliva might either be diluted with pool water or be contaminated by fodder fish, as frozen fish usually contains high levels of cortisol. In our study, we investigated how saliva cortisol levels are connected to blood cortisol levels and how saliva cortisol can be influenced by fodder fish. We examined saliva and blood samples in eleven bottlenose dolphins (Tursiops truncatus) kept in an outdoor and indoor facility in Germany. Furthermore, we assessed the cortisol levels of different kinds of fodder fish. Our data show that, although saliva cortisol values are elevated under stress and arousal, they seem not to be correlated with blood cortisol values. We also show that, after feeding, saliva cortisol values are increased up to 100-fold. Our results suggest that saliva cortisol measurements in dolphins have to be conducted and considered with care, as they can easily be contaminated. Moreover, it is important to use the right laboratory method in order to specifically detect cortisol; in our study, we conducted reliable tests, using LC-MS/MS.

Variation in plant leaf traits affects transmission and detectability of herbivore vibrational cues.

Many insects use plant-borne vibrations to obtain important information about their environment, such as where to find a mate or a prey, or when to avoid a predator. Plant species can differ in the way they vibrate, possibly affecting the reliability of information, and ultimately the decisions that are made by animals based on this information. We examined whether the production, transmission, and possible perception of plant-borne vibrational cues is affected by variation in leaf traits. We recorded vibrations of 69 Spodoptera exigua caterpillars foraging on four plant species that differed widely in their leaf traits (cabbage, beetroot, sunflower, and corn). We carried out a transmission and an airborne noise absorption experiment to assess whether leaf traits influence amplitude and frequency characteristics, and background noise levels of vibrational chewing cues. Our results reveal that species-specific leaf traits can influence transmission and potentially perception of herbivore-induced chewing vibrations. Experimentally-induced vibrations attenuated stronger on plants with thicker leaves. Amplitude and frequency characteristics of chewing vibrations measured near a chewing caterpillar were, however, not affected by leaf traits. Furthermore, we found a significant effect of leaf area, water content and leaf thickness-important plant traits against herbivory, on the vibrations induced by airborne noise. On larger leaves higher amplitude vibrations were induced, whereas on thicker leaves containing more water airborne noise induced higher peak frequencies. Our findings indicate that variation in leaf traits can be important for the transmission and possibly detection of vibrational cues.

Free-ranging bats combine three different cognitive processes for roost localization.

Animals have evolved different cognitive processes to localize crucial resources that are difficult to find. Relevant cognitive processes such as associative learning and spatial memory have commonly been studied in a foraging related context under controlled laboratory conditions. However, in natural environments, animals can use multiple cognitive processes to localize resources. In this field study, we used a pairwise choice experiment and automatic roost monitoring to assess how individually marked, free-ranging Bechstein's bats belonging to two different colonies use associative learning, spatial memory and social information when localizing suitable day roosts. To our knowledge, this study tests for the first time how associative learning, spatial memory and social information are used in the process of roost localization in bats under the natural conditions. We show that, when searching for new roosts, bats used associative learning to discriminate between suitable and unsuitable roosts. For re-localizing previously occupied roosts, bats used spatial memory rather than associative learning. Moreover, bats significantly improved the localization of suitable unfamiliar roosts and tended to increase their accuracy to re-localize previously occupied day roosts using social information. Our field experiments suggest that Bechstein's bats make hierarchical use of different cognitive processes when localizing day roosts. More generally, our study underlines that evaluating different cues under natural conditions is fundamental to understanding how natural selection has shaped the cognitive processes used for localizing resources.

Bioinspired sonar reflectors as guiding beacons for autonomous navigation.

Sonar sensors are universally applied in autonomous vehicles such as robots and driverless cars as they are inexpensive, energy-efficient, and provide accurate range measurements; however, they have some limitations. Their measurements can lead to ambiguous estimates and echo clutter can hamper target detection. In nature, echolocating bats experience similar problems when searching for food, especially if their food source is close to vegetation, as is the case for gleaning bats and nectar-feeding bats. However, nature has come up with solutions to overcome clutter problems and acoustically guide bats. Several bat-pollinated plants have evolved specially shaped floral parts that act as sonar reflectors, making the plants acoustically conspicuous. Here we show that artificial sonar beacons inspired by floral shapes streamline the navigation efficacy of sonar-guided robot systems. We developed floral-inspired reflector forms and demonstrate their functionality in 2 proof-of-principle experiments. First we show that the reflectors are easily recognized among dense clutter, and second we show that it is possible to discern different reflector shapes and use this identification to guide a robot through an unfamiliar environment. Bioinspired sonar reflectors could have a wide range of applications that could significantly advance sonar-guided systems.

Bats Actively Use Leaves as Specular Reflectors to Detect Acoustically Camouflaged Prey.

Filtering relevant signals from noisy sensory input is a crucial challenge for animals [1, 2]. Many bats are acoustic specialists relying on sound to find prey. They discern salient acoustic signals from irrelevant background masking noise. It has long been considered a sensory impossibility for bats to use solely echolocation for the detection of silent and motionless prey resting directly on foliage due to the masking effects of background echoes [3, 4]. Some bats, however, do successfully perform this seemingly impossible task [5], raising the question-what underlying acoustic and behavioral mechanisms do bats use to solve this conundrum? To address this question, we used biomimetic sonar to record high-resolution measurements of echoes from insects resting on leaves. Based on our echo recordings, we predicted optimal approach angles from which masking echoes can best be avoided. In behavioral experiments, we put these predictions to test. We recorded the prey approach behavior of wild bats in a flight cage equipped with an ultrasonic microphone synchronized with two high-speed cameras for 3D flightpath reconstructions. Bats approached prey from our predicted optimal oblique angles, using the leaf as a specular reflector to uncover previously acoustically hidden prey. Our findings disclose key behavioral and acoustic mechanisms enabling the detection of prey echoes that background clutter would otherwise mask. This work adds to the fundamental understanding of how bat echolocation strategies can override acoustic camouflage by silent, motionless prey, thus providing new insights into the evolutionary arms race between predators and their prey.

Classification of Sonar Targets in Air: A Neural Network Approach.

Ultrasonic sonar sensors are commonly used for contactless distance measurements in application areas such as automotive and mobile robotics. They can also be exploited to identify and classify sound-reflecting objects (targets), which may then be used as landmarks for navigation. In the presented work, sonar targets of different geometric shapes and sizes are classified with custom-engineered features. Artificial neural networks (ANNs) with multiple hidden layers are applied as classifiers and different features are tested as well as compared. We concentrate on features that are related to target strength estimates derived from pulse-compressed echoes. In doing so, one is able to distinguish different target geometries with a high rate of success and to perform tests with ANNs regarding their capabilities for size discrimination of targets with the same geometric shape. A comparison of achievable classifier performance with wideband and narrowband chirp excitation signals was conducted as well. The research indicates that our engineered features and excitation signals are suitable for the target classification task.

Acoustic communication in plant-animal interactions.

Acoustic communication is widespread and well-studied in animals but has been neglected in other organisms such as plants. However, there is growing evidence for acoustic communication in plant-animal interactions. While knowledge about active acoustic signalling in plants (i.e. active sound production) is still in its infancy, research on passive acoustic signalling (i.e. reflection of animal sounds) revealed that bat-dependent plants have adapted to the bats' echolocation systems by providing acoustic reflectors to attract their animal partners. Understanding the proximate mechanisms and ultimate causes of acoustic communication will shed light on an underestimated dimension of information transfer between plants and animals.

Sensory challenges for trawling bats: Finding transient prey on water surfaces.

Bats are able to identify obstacles and prey objects based exclusively on acoustic information acquired via echolocation. To assess the echo information potentially available to the trawling bat Noctilio leporinus, prey objects were ensonified with artificial bat calls and deduced echo target strengths (TS) of the reflected signals. The artificial calls consisted either of constant frequency (CF) or frequency modulated (FM) sounds. Detection distances were calculated for call intensities of N. leporinus emitted in the field and in confined space. Measurements of a transient target consisting of a brief water splash and subsequently expanding water ripples revealed that concentrically expanding water ripples can provide sufficiently loud echoes to be detected by trawling bats. Experiments with stationary targets revealed differences in TS depending on the type of signal used (CF or FM). A calculated maximum detection distance between 4.5 and 13.7 m for all measured targets indicates that prey detection in this very loud calling species occurs much earlier than suggested by estimations based on modifications in echolocation or flight behavior.

Bats Are Acoustically Attracted to Mutualistic Carnivorous Plants.

Mutualisms between plants and animals shape the world's ecosystems. In such interactions, achieving contact with the partner species is imperative. Plants regularly advertise themselves with signals that specifically appeal to the partner's perceptual preferences. For example, many plants have acquired traits such as brightly colored, fragrant flowers that attract pollinators with visual, olfactory, or--in the case of a few bat-pollinated flowers--even acoustic stimuli in the form of echo-reflecting structures. However, acoustic attraction in plants is rare compared to other advertisements and has never been found outside the pollination context and only in the Neotropics. We hypothesized that this phenomenon is more widespread and more diverse as plant-bat interactions also occur in the Paleotropics. In Borneo, mutualistic bats fertilize a carnivorous pitcher plant while roosting in its pitchers. The pitcher's orifice features a prolonged concave structure, which we predicted to distinctively reflect the bats' echolocation calls for a wide range of angles. This structure should facilitate the location and identification of pitchers even within highly cluttered surroundings. Pitchers lacking this structure should be less attractive for the bats. Ensonifications of the pitchers around their orifice revealed that this structure indeed acts as a multidirectional ultrasound reflector. In behavioral experiments where bats were confronted with differently modified pitchers, the reflector's presence clearly facilitated the finding and identification of pitchers. These results suggest that plants have convergently acquired reflectors in the Paleotropics and the Neotropics to acoustically attract bats, albeit for completely different ecological reasons.

Biosonar resolving power: echo-acoustic perception of surface structures in the submillimeter range.

The minimum distance for which two points still can be separated from each other defines the resolving power of a visual system. In an echo-acoustic context, the resolving power is usually measured as the smallest perceivable distance of two reflecting surfaces on the range axis and is found to be around half a millimeter for bats employing frequency modulated (FM) echolocation calls. Only few studies measured such thresholds with physical objects, most often bats were trained on virtual echoes i.e., echoes generated and played back by a computer; moreover, bats were sitting while they received the stimuli. In these studies differences in structure depth between 200 and 340 µm were found. However, these low thresholds were never verified for free-flying bats and real physical objects. Here, we show behavioral evidence that the echo-acoustic resolving power for surface structures in fact can be as low as measured for computer generated echoes and even lower, sometimes below 100 µm. We found this exceptional fine discrimination ability only when one of the targets showed spectral interferences in the frequency range of the bats' echolocation call while the other target did not. This result indicates that surface structure is likely to be perceived as a spectral quality rather than being perceived strictly in the time domain. Further, it points out that sonar resolving power directly depends on the highest frequency/shortest wavelength of the signal employed.

Floral acoustics: conspicuous echoes of a dish-shaped leaf attract bat pollinators.

The visual splendor of many diurnal flowers serves to attract visually guided pollinators such as bees and birds, but it remains to be seen whether bat-pollinated flowers have evolved analogous echo-acoustic signals to lure their echolocating pollinators. Here, we demonstrate how an unusual dish-shaped leaf displayed above the inflorescences of the vine Marcgravia evenia attracts bat pollinators. Specifically, this leaf's echoes fulfilled requirements for an effective beacon, that is, they were strong, multidirectional, and had a recognizable invariant echo signature. In behavioral experiments, presence of the leaves halved foraging time for flower-visiting bats.

High activity enables life on a high-sugar diet: blood glucose regulation in nectar-feeding bats.

High blood glucose levels caused by excessive sugar consumption are detrimental to mammalian health and life expectancy. Despite consuming vast quantities of sugar-rich floral nectar, nectar-feeding bats are long-lived, provoking the question of how they regulate blood glucose. We investigated blood glucose levels in nectar-feeding bats (Glossophaga soricina) in experiments in which we varied the amount of dietary sugar or flight time. Blood glucose levels increased with the quantity of glucose ingested and exceeded 25 mmol l(-1) blood in resting bats, which is among the highest values ever recorded in mammals fed sugar quantities similar to their natural diet. During normal feeding, blood glucose values decreased with increasing flight time, but only fell to expected values when bats spent 75 per cent of their time airborne. Either nectar-feeding bats have evolved mechanisms to avoid negative health effects of hyperglycaemia, or high activity is key to balancing blood glucose levels during foraging. We suggest that the coevolutionary specialization of bats towards a nectar diet was supported by the high activity and elevated metabolic rates of these bats. High activity may have conferred benefits to the bats in terms of behavioural interactions and foraging success, and is simultaneously likely to have increased their efficiency as plant pollinators.

Size discrimination of hollow hemispheres by echolocation in a nectar feeding bat.

Nectar feeding bats use echolocation to find their flowers in the dense growth of tropical rainforests, and such flowers have evolved acoustic features that make their echo more conspicuous to their pollinators. To shed light on the sensory and cognitive basis of echoacoustic object recognition we conducted a size discrimination experiment with the nectarivorous bat Glossophaga soricina and compared the bats' behavioural performance with the echoic features of the training objects. We chose a simple geometric form, the hollow hemisphere, as the training object because of its resemblance to the bell-shaped concave form of many bat flowers, as well as its special acoustic qualities. The hemispheres showed a characteristic echo pattern, which was constant over a wide range of angles of sound incidence. We found systematic size-dependent changes in the echo's temporal and spectral pattern as well as in amplitude. Bats were simultaneously confronted with seven different sizes of hollow hemispheres presented from their concave sides. Visits to one particular size were rewarded with sugar water, while we recorded the frequency of visits to the unrewarded hemispheres. We found that: (1) bats learned to discriminate between hemispheres of different size with ease; (2) the minimum size difference for discrimination was a constant percentage of the hemisphere's size (Weber fraction: approximately 16% of the radius); (3) the comparison of behavioural data and impulse response measurements of the objects' echoes yielded discrimination thresholds for mean intensity differences (1.3 dB), the temporal pattern (3-22 micros) and the change of spectral notch frequency (approximately 16%). We discuss the advantages of discrimination in the frequency and/or time domain.