Understanding park visitors' soundscape perception using subjective and objective measurement.

Environmental noise knows no boundaries, affecting even protected areas. Noise pollution, originating from both external and internal sources, imposes costs on these areas. It is associated with adverse health effects, while natural sounds contribute to cognitive and emotional improvements as ecosystem services. When it comes to parks, individual visitors hold unique perceptions of soundscapes, which can be shaped by various factors such as their motivations for visiting, personal norms, attitudes towards specific sounds, and expectations. In this study, we utilized linear models and geospatial data to evaluate how visitors' personal norms and attitudes, the park's acoustic environment, visitor counts, and the acoustic environment of visitors' neighborhoods influenced their perception of soundscapes at Muir Woods National Monument. Our findings indicate that visitors' subjective experiences had a greater impact on their perception of the park's soundscape compared to purely acoustic factors like sound level of the park itself. Specifically, we found that motivations to hear natural sounds, interference caused by noise, sensitivity to noise, and the sound levels of visitors' home neighborhoods influenced visitors' perception of the park's soundscape. Understanding how personal factors shape visitors' soundscape perception can assist urban and non-urban park planners in effectively managing visitor experiences and expectations.

Effects of wind turbine noise on songbird behavior during nonbreeding season.

Anthropogenic noise is one of the fastest growing, globally widespread pollutants, affecting countless species worldwide. Despite accumulating evidence of the negative impacts of wind turbines on wildlife, little is known about how the noise they generate affects ecological systems. Songbirds may be susceptible to noise pollution due to their reliance on vocal communication and thus, in this field study, we examined how songbirds are affected by wind turbine noise. We broadcasted noise produced by one wind turbine in a migratory stopover site during the nonbreeding season. Throughout the study, we repeatedly monitored the acoustic environment and songbird community before, during, and after the noise treatments with passive acoustic monitoring and mist netting. We employed generalized linear mixed effects models to assess the impact of experimental noise treatment on birds behavior and likelihood ratio tests to compare models with variables of interest with null models. The daily number of birds in the presence of wind turbine noise decreased by approximately 30% compared with the before and after phases. This reduction had a significant spatial pattern; the largest decrease was closer to the speaker and on its downwind side, fitting measured sound propagation. Although we found no impact on species diversity, two out of three most common species showed clear avoidance behavior: 45% and 36% decrease in abundance for the lesser whitethroat (Sylvia curruca) and Sardinian warbler (Sylvia melanocephala momus), respectively. In the after phase, there were lingering effects on the lesser whitethroat. The age structure of the lesser whitethroat population was affected because only juvenile birds showed avoidance behavior. No difference in avoidance extent was found between migratory and nonmigratory species, but the impacts of displacement on migrants during stopover are especially troubling from a conservation perspective. Our results stress the need to address the impacts of noise pollution on wildlife when planning noise-generating infrastructures, such as wind turbines, to allow for sustainable development without threatening already declining songbird populations.

El ruido antropogénico es uno de los contaminantes con mayor crecimiento y distribución a nivel mundial, por lo que afecta a incontables especies en todo el mundo. A pesar de acumular evidencia sobre el impacto negativo que tienen las turbinas eólicas sobre la fauna, se sabe muy poco sobre cómo el ruido que generan afecta a los sistemas ecológicos. Las aves canoras pueden ser susceptibles a la contaminación sonora ya que dependen de la comunicación vocal y, por lo tanto, en este estudio de campo, analizamos cómo les afecta el sonido producido por las turbinas eólicas. Transmitimos ruido producido por una turbina en un punto de parada migratorio durante la temporada no reproductiva. Durante el estudio, monitoreamos repetidas veces el entorno acústico y la comunidad de aves canoras antes, durante y después de los tratamientos de ruido con monitoreo acústico pasivo y redes de niebla. Empleamos modelos de efectos lineales mixtos generalizados para evaluar el impacto del ruido experimental sobre el comportamiento de las aves y pruebas de probabilidad de proporción para comparar los modelos con variables de interés con los modelos nulos. El número diario de aves en la presencia del ruido de turbinas eólicas disminuyó aproximadamente un 30% en comparación con las fases de antes y después. Esta reducción tuvo un patrón espacial significativo: la mayor disminución ocurrió más cerca a la bocina y en el lado de sotavento, lo que se ajusta a la medida de la propagación del sonido. Aunque no encontramos impacto alguno sobre la diversidad de especies, dos de tres de las especies más comunes mostraron un comportamiento de evasión evidente: 45% y 36% de disminución en la abundancia de Sylvia curruca y Sylvia melanocephala momus, respectivamente. Durante la fase posterior al ruido, observamos efectos prolongados en S. curruca. La composición de edades de la población de S. curruca se vio afectada porque sólo los individuos juveniles mostraron un comportamiento de evasión. No encontramos una diferencia en el grado de evasión entre las especies migratorias y no migratorias, pero el impacto del traslado sobre las migrantes durante el punto de parada es de preocupación especial desde una perspectiva de conservación. Nuestros resultados acentúan la necesidad de abordar el impacto de la contaminación sonora sobre la fauna cuando se planean estructuras que producen ruido, como las turbinas eólicas, para permitir el desarrollo sustentable sin amenazar a las poblaciones de aves canoras que ya están en declive. Efectos del ruido de turbinas eólicas sobre el comportamiento de las aves canoras durante la temporada no reproductiva.

EcoCountHelper: an R package and analytical pipeline for the analysis of ecological count data using GLMMs, and a case study of bats in Grand Teton National Park.

Here we detail the use of an R package, 'EcoCountHelper', and an associated analytical pipeline aimed at making generalized linear mixed-effects model (GLMM)-based analysis of ecological count data more accessible. We recommend a GLMM-based analysis workflow that allows the user to (1) employ selection of distributional forms (Poisson vs negative binomial) and zero-inflation (ZIP and ZINB, respectively) using AIC and variance-mean plots, (2) examine models for goodness-of-fit using simulated residual diagnostics, (3) interpret model results via easy to understand outputs of changes in predicted responses, and (4) compare the magnitude of predictor variable effects via effects plots. Our package uses a series of easy-to-use functions that can accept both wide- and long-form multi-taxa count data without the need for programming experience. To demonstrate the utility of this approach, we use our package to model acoustic bat activity data relative to multiple landscape characteristics in a protected area (Grand Teton National Park), which is threatened by encroaching disease-white nose syndrome. Global threats to bat conservation such as disease and deforestation have prompted extensive research to better understand bat ecology. Notwithstanding these efforts, managers operating on lands crucial to the persistence of bat populations are often equipped with too little information regarding local bat activity to make informed land-management decisions. In our case study in the Tetons, we found that an increased prevalence of porous buildings increases activity levels of Eptesicus fuscus and Myotis volans; Myotis lucifugus activity decreases as distance to water increases; and Myotis volans activity increases with the amount of forested area. By using GLMMs in tandem with 'EcoCountHelper', managers without advanced programmatic or statistical expertise can assess the effects of landscape characteristics on wildlife in a statistically-robust framework.

Anti-bat ultrasound production in moths is globally and phylogenetically widespread.

Warning signals are well known in the visual system, but rare in other modalities. Some moths produce ultrasonic sounds to warn bats of noxious taste or to mimic unpalatable models. Here, we report results from a long-term study across the globe, assaying moth response to playback of bat echolocation. We tested 252 genera, spanning most families of large-bodied moths, and document anti-bat ultrasound production in 52 genera, with eight subfamily origins described. Based on acoustic analysis of ultrasonic emissions and palatability experiments with bats, it seems that acoustic warning and mimicry are the raison d'être for sound production in most moths. However, some moths use high-duty-cycle ultrasound capable of jamming bat sonar. In fact, we find preliminary evidence of independent origins of sonar jamming in at least six subfamilies. Palatability data indicate that jamming and warning are not mutually exclusive strategies. To explore the possible organization of anti-bat warning sounds into acoustic mimicry rings, we intensively studied a community of moths in Ecuador and, using machine-learning approaches, found five distinct acoustic clusters. While these data represent an early understanding of acoustic aposematism and mimicry across this megadiverse insect order, it is likely that ultrasonically signaling moths comprise one of the largest mimicry complexes on earth.

A stochastic simulation model for assessing the masking effects of road noise for wildlife, outdoor recreation, and bioacoustic monitoring.

Traffic noise is one of the leading causes of reductions in animal abundances near roads. Acoustic masking of conspecific signals and adventitious cues is one mechanism that likely causes animals to abandon loud areas. However, masking effects can be difficult to document in situ and the effects of infrequent noise events may be impractical to study. Here, we present the Soundscapes model, a stochastic individual-based model that dynamically models the listening areas of animals searching for acoustic resources ("searchers"). The model also studies the masking effects of noise for human detections of the searchers. The model is set in a landscape adjacent to a road. Noise produced by vehicles traveling on that road is represented by calibrated spectra that vary with speed. Noise propagation is implemented using ISO-9613 procedures. We present demonstration simulations that quantify declines in searcher efficiency and human detection of searchers at relatively low traffic volumes, fewer than 50 vehicles per hour. Traffic noise is pervasive, and the Soundscapes model offers an extensible tool to study the effects of noise on bioacoustics monitoring, point-count surveys, the restorative value of natural soundscapes, and auditory performance in an ecological context.

Experimentally broadcast ocean surf and river noise alters birdsong.

Anthropogenic noise and its effects on acoustic communication have received considerable attention in recent decades. Yet, the natural acoustic environment's influence on communication and its role in shaping acoustic signals remains unclear. We used large-scale playbacks of ocean surf in coastal areas and whitewater river noise in riparian areas to investigate how natural sounds influences song structure in six songbird species. We recorded individuals defending territories in a variety of acoustic conditions across 19 study sites in California and 18 sites in Idaho. Acoustic characteristics across the sites included naturally quiet 'control' sites, 'positive control' sites that were adjacent to the ocean or a whitewater river and thus were naturally noisy, 'phantom' playback sites that were exposed to continuous broadcast of low-frequency ocean surf or whitewater noise, and 'shifted' playback sites with continuous broadcast of ocean surf or whitewater noise shifted up in frequency. We predicted that spectral and temporal song structure would generally correlate with background sound amplitude and that signal features would differ across site types based on the spectral profile of the acoustic environment. We found that the ways in which song structure varied with background acoustics were quite variable from species to species. For instance, in Idaho both the frequency bandwidth and duration of lazuli bunting (Passerina amoena) and song sparrow (Melospiza melodia) songs decreased with elevated background noise, but these song features were unrelated to background noise in the warbling vireo (Vireo gilvus), which tended to increase both the minimum and maximum frequency of songs with background noise amplitude. In California, the bandwidth of the trill of white-crowned sparrow (Zonotrichia leucophrys) song decreased with background noise amplitude, matching results of previous studies involving both natural and anthropogenic noise. In contrast, wrentit (Chamaea fasciata) song bandwidth was positively related to the amplitude of background noise. Although responses were quite heterogeneous, song features of all six species varied with amplitude and/or frequency of background noise. Collectively, these results provide strong evidence that natural soundscapes have long influenced vocal behavior. More broadly, the evolved behavioral responses to the long-standing challenges presented by natural sources of noise likely explain the many responses observed for species communicating in difficult signal conditions presented by human-made noise.

Hidden Phylogenomic Signal Helps Elucidate Arsenurine Silkmoth Phylogeny and the Evolution of Body Size and Wing Shape Trade-Offs.

One of the key objectives in biological research is understanding how evolutionary processes have produced Earth's diversity. A critical step toward revealing these processes is an investigation of evolutionary tradeoffs-that is, the opposing pressures of multiple selective forces. For millennia, nocturnal moths have had to balance successful flight, as they search for mates or host plants, with evading bat predators. However, the potential for evolutionary trade-offs between wing shape and body size are poorly understood. In this study, we used phylogenomics and geometric morphometrics to examine the evolution of wing shape in the wild silkmoth subfamily Arsenurinae (Saturniidae) and evaluate potential evolutionary relationships between body size and wing shape. The phylogeny was inferred based on 782 loci from target capture data of 42 arsenurine species representing all 10 recognized genera. After detecting in our data one of the most vexing problems in phylogenetic inference-a region of a tree that possesses short branches and no "support" for relationships (i.e., a polytomy), we looked for hidden phylogenomic signal (i.e., inspecting differing phylogenetic inferences, alternative support values, quartets, and phylogenetic networks) to better illuminate the most probable generic relationships within the subfamily. We found there are putative evolutionary trade-offs between wing shape, body size, and the interaction of fore- and hindwing (HW) shape. Namely, body size tends to decrease with increasing HW length but increases as forewing (FW) shape becomes more complex. Additionally, the type of HW (i.e., tail or no tail) a lineage possesses has a significant effect on the complexity of FW shape. We outline possible selective forces driving the complex HW shapes that make Arsenurinae, and silkmoths as a whole, so charismatic. [Anchored hybrid enrichment; Arsenurinae; geometric morphometrics; Lepidoptera; phylogenomics; Saturniidae.].

Adaptive shifts underlie the divergence in wing morphology in bombycoid moths.

The evolution of flapping flight is linked to the prolific success of insects. Across Insecta, wing morphology diversified, strongly impacting aerodynamic performance. In the presence of ecological opportunity, discrete adaptive shifts and early bursts are two processes hypothesized to give rise to exceptional morphological diversification. Here, we use the sister-families Sphingidae and Saturniidae to answer how the evolution of aerodynamically important traits is linked to clade divergence and through what process(es) these traits evolve. Many agile Sphingidae evolved hover feeding behaviours, while adult Saturniidae lack functional mouth parts and rely on a fixed energy budget as adults. We find that Sphingidae underwent an adaptive shift in wing morphology coincident with life history and behaviour divergence, evolving small high aspect ratio wings advantageous for power reduction that can be moved at high frequencies, beneficial for flight control. By contrast, Saturniidae, which do not feed as adults, evolved large wings and morphology which surprisingly does not reduce aerodynamic power, but could contribute to their erratic flight behaviour, aiding in predator avoidance. We suggest that after the evolution of flapping flight, diversification of wing morphology can be potentiated by adaptative shifts, shaping the diversity of wing morphology across insects.

Assessing the Vulnerabilities of Vertebrate Species to Light and Noise Pollution: Expert Surveys Illuminate the Impacts on Specialist Species.

Global expansion of lighting and noise pollution alters how animals receive and interpret environmental cues. However, we lack a cross-taxon understanding of how animal traits influence species vulnerability to this growing phenomenon. This knowledge is needed to improve the design and implementation of policies that mitigate or reduce sensory pollutants. We present results from an expert knowledge survey that quantified the relative influence of 21 ecological, anatomical, and physiological traits on the vulnerability of terrestrial vertebrates to elevated levels of anthropogenic lighting and noise. We aimed not only to quantify the importance of threats and the relative influence of traits as viewed by sensory and wildlife experts, but to examine knowledge gaps based on the variation in responses. Identifying traits that had less consensus can guide future research for strengthening ecologists' and conservation biologists' understanding of sensory abilities. Our findings, based on 280 responses of expert opinion, highlight the increasing recognition among experts that sensory pollutants are important to consider in management and conservation decisions. Participant responses show mounting threats to species with narrow niches; especially habitat specialists, nocturnal species, and those with the greatest ability to differentiate environmental visual and auditory cues. Our results call attention to the threat specialist species face and provide a generalizable understanding of which species require additional considerations when developing conservation policies and mitigation strategies in a world altered by expanding sensory pollutant footprints. We provide a step-by-step example for translating these results to on-the-ground conservation planning using two species as case studies.

Artificial night light and anthropogenic noise interact to influence bird abundance over a continental scale.

The extent of artificial night light and anthropogenic noise (i.e., "light" and "noise") impacts is global and has the capacity to threaten species across diverse ecosystems. Existing research involving impacts of light or noise has primarily focused on noise or light alone and single species; however, these stimuli often co-occur and little is known about how co-exposure influences wildlife and if and why species may vary in their responses. Here, we had three aims: (1) to investigate species-specific responses to light, noise, and the interaction between the two using a spatially explicit approach to model changes in abundance of 140 prevalent bird species across North America, (2) to investigate responses to the interaction between light exposure and night length, and (3) to identify functional traits and habitat affiliations that explain variation in species-specific responses to these sensory stimuli with phylogenetically informed models. We found species that responded to noise exposure generally decreased in abundance, and the additional presence of light interacted synergistically with noise to exacerbate its negative effects. Moreover, the interaction revealed negative emergent responses for several species that only reacted when light and noise co-occurred. Additionally, an interaction between light and night length revealed 47 species increased in abundance with light exposure during longer nights. In addition to modifying behavior with optimal temperature and potential foraging opportunities, birds might be attracted to light, yet suffer inadvertent physiological consequences. The trait that most strongly related to avian response to light and noise was habitat affiliation. Specifically, species that occupy closed habitat were less tolerant of both sensory stressors compared to those that occupy open habitat. Further quantifying the contexts and intrinsic traits that explain how species respond to noise and light will be fundamental to understanding the ecological consequences of a world that is ever louder and brighter.

Noise distracts foraging bats.

Predators frequently must detect and localize their prey in challenging environments. Noisy environments have been prevalent across the evolutionary history of predator-prey relationships, but now with increasing anthropogenic activities noise is becoming a more prominent feature of many landscapes. Here, we use the gleaning pallid bat, Antrozous pallidus, to investigate the mechanism by which noise disrupts hunting behaviour. Noise can primarily function to mask-obscure by spectrally overlapping a cue of interest, or distract-occupy an animal's attentional or other cognitive resources. Using band-limited white noise treatments that either overlapped the frequencies of a prey cue or did not overlap this cue, we find evidence that distraction is a primary driver of reduced hunting efficacy in an acoustically mediated predator. Under exposure to both noise types successful prey localization declined by half, search time nearly tripled, and bats used 25% more sonar pulses than when hunting in ambient conditions. Overall, the pallid bat does not seem capable of compensating for environmental noise. These findings have implications for mitigation strategies, specifically the importance of reducing sources of noise on the landscape rather than attempting to reduce the bandwidth of anthropogenic noise.

The phantom chorus: birdsong boosts human well-being in protected areas.

Spending time in nature is known to benefit human health and well-being, but evidence is mixed as to whether biodiversity or perceptions of biodiversity contribute to these benefits. Perhaps more importantly, little is known about the sensory modalities by which humans perceive biodiversity and obtain benefits from their interactions with nature. Here, we used a 'phantom birdsong chorus' consisting of hidden speakers to experimentally increase audible birdsong biodiversity during 'on' and 'off' (i.e. ambient conditions) blocks on two trails to study the role of audition in biodiversity perception and self-reported well-being among hikers. Hikers exposed to the phantom chorus reported higher levels of restorative effects compared to those that experienced ambient conditions on both trails; however, increased restorative effects were directly linked to the phantom chorus on one trail and indirectly linked to the phantom chorus on the other trail through perceptions of avian biodiversity. Our findings add to a growing body of evidence linking mental health to nature experiences and suggest that audition is an important modality by which natural environments confer restorative effects. Finally, our results suggest that maintaining or improving natural soundscapes within protected areas may be an important component to maximizing human experiences.

Sensory pollutants alter bird phenology and fitness across a continent.

Expansion of anthropogenic noise and night lighting across our planet1,2 is of increasing conservation concern3-6. Despite growing knowledge of physiological and behavioural responses to these stimuli from single-species and local-scale studies, whether these pollutants affect fitness is less clear, as is how and why species vary in their sensitivity to these anthropic stressors. Here we leverage a large citizen science dataset paired with high-resolution noise and light data from across the contiguous United States to assess how these stimuli affect reproductive success in 142 bird species. We find responses to both sensory pollutants linked to the functional traits and habitat affiliations of species. For example, overall nest success was negatively correlated with noise among birds in closed environments. Species-specific changes in reproductive timing and hatching success in response to noise exposure were explained by vocalization frequency, nesting location and diet. Additionally, increased light-gathering ability of species' eyes was associated with stronger advancements in reproductive timing in response to light exposure, potentially creating phenological mismatches7. Unexpectedly, better light-gathering ability was linked to reduced clutch failure and increased overall nest success in response to light exposure, raising important questions about how responses to sensory pollutants counteract or exacerbate responses to other aspects of global change, such as climate warming. These findings demonstrate that anthropogenic noise and light can substantially affect breeding bird phenology and fitness, and underscore the need to consider sensory pollutants alongside traditional dimensions of the environment that typically inform biodiversity conservation.

Why conservation biology can benefit from sensory ecology.

Global expansion of human activities is associated with the introduction of novel stimuli, such as anthropogenic noise, artificial lights and chemical agents. Progress in documenting the ecological effects of sensory pollutants is weakened by sparse knowledge of the mechanisms underlying these effects. This severely limits our capacity to devise mitigation measures. Here, we integrate knowledge of animal sensory ecology, physiology and life history to articulate three perceptual mechanisms-masking, distracting and misleading-that clearly explain how and why anthropogenic sensory pollutants impact organisms. We then link these three mechanisms to ecological consequences and discuss their implications for conservation. We argue that this framework can reveal the presence of 'sensory danger zones', hotspots of conservation concern where sensory pollutants overlap in space and time with an organism's activity, and foster development of strategic interventions to mitigate the impact of sensory pollutants. Future research that applies this framework will provide critical insight to preserve the natural sensory world.

Time of night and moonlight structure vertical space use by insectivorous bats in a Neotropical rainforest: an acoustic monitoring study.

BACKGROUND: Previous research has shown diverse vertical space use by various taxa, highlighting the importance of forest vertical structure. Yet, we know little about vertical space use of tropical forests, and we often fail to explore how this three-dimensional space use changes over time. METHODS: Here we use canopy tower systems in French Guiana and passive acoustic monitoring to measure Neotropical bat activity above and below the forest canopy throughout nine nights. We use a Bayesian generalized linear mixed effect model and kernel density estimates to demonstrate patterns in space-use over time. RESULTS: We found that different bats use both canopy and understory space differently and that these patterns change throughout the night. Overall, bats were more active above the canopy (including Cormura brevirostris, Molossus molossus, Peropteryx kappleri and Peropteryx macrotis), but multiple species or acoustic complexes (when species identification was impossible) were more active in the understory (such as Centronycteris maximiliani, Myotis riparius, Pteronotus alitonus and Pteronotus rubiginosus). We also found that most bats showed temporally-changing preferences in hourly activity. Some species were less active (e.g., P. kappleri and P. macrotis), whereas others were more active (Pteronotus gymnonotus, C. brevirostris, and M. molossus) on nights with higher moon illuminance. DISCUSSION: Here we show that Neotropical bats use habitat above the forest canopy and within the forest understory differently throughout the night. While bats generally were more active above the forest canopy, we show that individual groups of bats use space differently over the course of a night, and some prefer the understory. This work highlights the need to consider diel cycles in studies of space use, as animals use different habitats during different periods of the day.

Phylogenomics reveals the evolutionary timing and pattern of butterflies and moths.

Butterflies and moths (Lepidoptera) are one of the major superradiations of insects, comprising nearly 160,000 described extant species. As herbivores, pollinators, and prey, Lepidoptera play a fundamental role in almost every terrestrial ecosystem. Lepidoptera are also indicators of environmental change and serve as models for research on mimicry and genetics. They have been central to the development of coevolutionary hypotheses, such as butterflies with flowering plants and moths' evolutionary arms race with echolocating bats. However, these hypotheses have not been rigorously tested, because a robust lepidopteran phylogeny and timing of evolutionary novelties are lacking. To address these issues, we inferred a comprehensive phylogeny of Lepidoptera, using the largest dataset assembled for the order (2,098 orthologous protein-coding genes from transcriptomes of 186 species, representing nearly all superfamilies), and dated it with carefully evaluated synapomorphy-based fossils. The oldest members of the Lepidoptera crown group appeared in the Late Carboniferous (∼300 Ma) and fed on nonvascular land plants. Lepidoptera evolved the tube-like proboscis in the Middle Triassic (∼241 Ma), which allowed them to acquire nectar from flowering plants. This morphological innovation, along with other traits, likely promoted the extraordinary diversification of superfamily-level lepidopteran crown groups. The ancestor of butterflies was likely nocturnal, and our results indicate that butterflies became day-flying in the Late Cretaceous (∼98 Ma). Moth hearing organs arose multiple times before the evolutionary arms race between moths and bats, perhaps initially detecting a wide range of sound frequencies before being co-opted to specifically detect bat sonar. Our study provides an essential framework for future comparative studies on butterfly and moth evolution.

An improved method for utilizing high-throughput amplicon sequencing to determine the diets of insectivorous animals.

DNA analysis of predator faeces using high-throughput amplicon sequencing (HTS) enhances our understanding of predator-prey interactions. However, conclusions drawn from this technique are constrained by biases that occur in multiple steps of the HTS workflow. To better characterize insectivorous animal diets, we used DNA from a diverse set of arthropods to assess PCR biases of commonly used and novel primer pairs for the mitochondrial gene, cytochrome oxidase C subunit 1 (COI). We compared diversity recovered from HTS of bat guano samples using a commonly used primer pair "ZBJ" to results using the novel primer pair "ANML." To parameterize our bioinformatics pipeline, we created an arthropod mock community consisting of single-copy (cloned) COI sequences. To examine biases associated with both PCR and HTS, mock community members were combined in equimolar amounts both pre- and post-PCR. We validated our system using guano from bats fed known diets and using composite samples of morphologically identified insects collected in pitfall traps. In PCR tests, the ANML primer pair amplified 58 of 59 arthropod taxa (98%), whereas ZBJ amplified 24-40 of 59 taxa (41%-68%). Furthermore, in an HTS comparison of field-collected samples, the ANML primers detected nearly fourfold more arthropod taxa than the ZBJ primers. The additional arthropods detected include medically and economically relevant insect groups such as mosquitoes. Results revealed biases at both the PCR and sequencing levels, demonstrating the pitfalls associated with using HTS read numbers as proxies for abundance. The use of an arthropod mock community allowed for improved bioinformatics pipeline parameterization.

Fireflies thwart bat attack with multisensory warnings.

Many defended animals prevent attacks by displaying warning signals that are highly conspicuous to their predators. We hypothesized that bioluminescing fireflies, widely known for their vibrant courtship signals, also advertise their noxiousness to echolocating bats. To test this postulate, we pit naïve big brown bats (Eptesicus fuscus) against chemically defended fireflies (Photinus pyralis) to examine whether and how these beetles transmit salient warnings to bats. We demonstrate that these nocturnal predators learn to avoid noxious fireflies using either vision or echolocation and that bats learn faster when integrating information from both sensory streams-providing fundamental evidence that multisensory integration increases the efficacy of warning signals in a natural predator-prey system. Our findings add support for a warning signal origin of firefly bioluminescence and suggest that bat predation may have driven evolution of firefly bioluminescence.

The evolution of anti-bat sensory illusions in moths.

Prey transmit sensory illusions to redirect predatory strikes, creating a discrepancy between what a predator perceives and reality. We use the acoustic arms race between bats and moths to investigate the evolution and function of a sensory illusion. The spinning hindwing tails of silk moths (Saturniidae) divert bat attack by reflecting sonar to create a misleading echoic target. We characterized geometric morphometrics of moth hindwings across silk moths, mapped these traits onto a new, robust phylogeny, and found that elaborated hindwing structures have converged on four adaptive shape peaks. To test the mechanism underlying these anti-bat traits, we pit bats against three species of silk moths with experimentally altered hindwings that created a representative gradient of ancestral and extant hindwing shapes. High-speed videography of battles reveals that moths with longer hindwings and tails more successfully divert bat attack. We postulate that sensory illusions are widespread and are underappreciated drivers of diversity across systems.