The Zebrafish Information Network (ZFIN, zfin.org) is the database resource for genetic, genomic, and phenotypic data from research using zebrafish, Danio rerio. ZFIN curates information about genetic perturbations, gene expression, phenotype, gene function, and human disease models from zebrafish research publications and makes these data available to researchers worldwide. Over the past 20 years, zebrafish have increasingly been used to investigate the effects of environmental exposures, becoming an ideal model to study toxicity, phenotypic outcomes, and gene-chemical interactions. Despite this, database resources supporting zebrafish toxicology and environmental exposure research are limited. To fill this gap, ZFIN has expanded functionality to incorporate and convey toxicology data better. ZFIN annotations for gene expression, phenotype, and human disease models include information about genotypes and experimental conditions used. One type of experimental condition the database captures is the application of chemicals to zebrafish. ZFIN annotates chemicals using the Chemical Entities of Biological Interest Ontology (ChEBI) along with the Zebrafish Experimental Conditions Ontology (ZECO) to denote route of exposure and other experimental conditions. These features allow researchers to search phenotypes and human disease models linked to chemicals more efficiently. Here we discuss how experimental conditions are displayed on ZFIN web pages, the data displayed on chemical term pages, and how to search and download data associated with chemical exposure experiments.

Decision making has important fitness consequences in domains as varied as foraging, mating and predator avoidance. In noisy ecosystems, animals often inform their decisions via information streams that may span sensory modalities. Despite theoretical predictions, we have little understanding of whether attending to multiple cues enhances decision-making performance or reduces it by creating cognitive overload. The predatory fringe-lipped bat, Trachops cirrhosus, hunts by eavesdropping on auditory prey cues, but it utilizes information from other sensory modalities when challenged in the laboratory and rapidly adopts new hunting behaviours through social learning. When hunting frogs, it uses their calls to identify palatable species. T & uacute;ngara frog, Engystomops pustulosus, males court females with a multimodal display: their calls (auditory) and the simultaneously inflating vocal sac (visual) create water ripples (seismic). Here we evaluated decision making in fringe-lipped bats attending to ambiguous stimuli with and without additional cues. We quantified foraging decisions in response to synthesized hybrid calls spanning a gradient from calls of the palatable t & uacute;ngara frog to those of a poisonous toad. Hybrid calls were presented to bats in silence and with additional stimuli: social cues (conspecific chewing noises), environmental noise (anuran chorus) and additional cues associated with target frog calls (vocal sac motion and water ripples). We predicted (1) social and target cues would make hybrid calls more attractive as a potential food source and (2) decision making would be slower in the presence of noise. Counter to expectation, bats maintained highly consistent responses to hybrid stimuli regardless of treatment, supporting neither the enhanced performance hypothesis nor the cognitive overload hypothesis on decision making. We observed flexibility in minimum approach thresholds, however. Bats showed greater exploratory and risk-taking behaviour when given access to information in the form of target cues in noise. Our results elucidate how cognitively complex animals deal with information complexity in varying decision-making contexts. (c) 2025 The Authors. Published by Elsevier Ltd on behalf of The Association for the Study of Animal Behaviour. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/li censes/by-nc-nd/4.0/).

Across many taxa, males gather in leks to perform multisensory courtship displays for females. At a lek, changes in the sensory scene over the course of mate evaluation are inevitable. This dynamic nature makes a female’s ability to recall the location of individual signalers an important component of female mate choice. It is hypothesized that complex (especially multimodal) displays may improve a female’s ability to remember and thus discriminate amongst potential mates. To test this hypothesis, we presented female túngara frogs ( Physalaemus (= Engystomops ) pustulosus ) with male calls (auditory) and robotic frogs (visual) that could later be obstructed from view via the lowering of blinds. Specifically, we asked if the visual component of a multimodal display improves the ability of a receiver to remember a signaler. While our initial experiment did not find memory instantiation in female frogs, subsequent experiments with a longer presentation time and a brief period of silence successfully instantiated memories in females. Moreover, females continued to demonstrate significant preferences for calls associated with the visual cue even after 25 s following the obstruction of the visual stimulus (robotic frog). Thus, we propose that the duration of presentation and/or the silent period impact memory capability. Silence is common in choruses, and our data suggest that complex, multisensory stimuli may have evolved to help females remember their preferred mate even with such fluctuations in signal production. LAY SUMMARY Multisensory signals function to stimulate one or more of a receiver’s senses, and in doing so, may make signalers more memorable to receivers. Multisensory signals are common in mating displays, including the advertisement of a male túngara frog: a call (acoustic) and a vocal sac inflation (visual). Here, we demonstrate that female túngara frogs were more likely to remember a multisensory male display over a unisensory display, but only after fluctuations in the signal’s intensity. ### Competing Interest Statement The authors have declared no competing interest. National Science Foundation, https://ror.org/021nxhr62, IOS-1914646

Extreme climatic events, such as hurricanes, droughts, and floods, have been implicated in altering various phenotypic traits in affected populations. The 2015 El Niño cycle led to one of the driest wet seasons recorded throughout parts of Central America. We examined the effects of this climatic event on sexual and natural selection using behavioral and morphological data from túngara frogs (Engystomops pustulosus) collected before, during, and after this event. In a species renowned for robust female mating preferences, we found a cross-generational reduction and loss of female mate preferences for certain male advertisement call properties following this event. We identified reductions in body length and simultaneous increases in condition during the climatic event. Reduced availability and permanence of aquatic breeding sites may explain these behavioral and morphological changes. Mate choice preferences reverted rapidly, while the restoration of body length was sex dependent following this period of drastically reduced precipitation. These findings highlight immediate and lingering effects of single climatic events and demonstrate the ability of climatic events to disrupt established mate preferences. As climate change increases the frequency and severity of extreme events, monitoring behavioral and morphological phenotypes will be imperative to fully understand the consequences of these events on animal populations.

Danio rerio is a model organism used to investigate vertebrate development. Manipulation of the zebrafish genome and resultant gene products by mutation or targeted knockdown has made the zebrafish a good system for investigating gene function, providing a resource to investigate genetic contributors to phenotype and human disease. Phenotypic outcomes can be the result of gene mutation, targeted knockdown of gene products, manipulation of experimental conditions, or any combination thereof. Zebrafish have been used in various genetic and chemical screens to identify genetic and environmental contributors to phenotype and disease outcomes. The Zebrafish Information Network (ZFIN) is the central repository for genetic, genomic, and phenotypic data that result from research using Danio rerio . Here we describe how ZFIN annotates phenotype, expression, and disease model data across various experimental designs, how we computationally determine wild-type gene expression, the phenotypic gene, and how these results allow us to propagate gene expression, phenotype, and disease model data to the correct gene, or gene related entity.

Background Infectious diseases of farmed and wild animals pose a recurrent threat to food security and human health. The macrophage, a key component of the innate immune system, is the first line of defence against many infectious agents and plays a major role in shaping the adaptive immune response. However, this phagocyte is a target and host for many pathogens. Understanding the molecular basis of interactions between macrophages and pathogens is therefore crucial for the development of effective strategies to combat important infectious diseases. Results We explored how porcine pluripotent stem cells (PSCs) can provide a limitless in vitro supply of genetically and experimentally tractable macrophages. Porcine PSC-derived macrophages (PSCdMs) exhibited molecular and functional characteristics of ex vivo primary macrophages and were productively infected by pig pathogens, including porcine reproductive and respiratory syndrome virus (PRRSV) and African swine fever virus (ASFV), two of the most economically important and devastating viruses in pig farming. Moreover, porcine PSCdMs were readily amenable to genetic modification by CRISPR/Cas9 gene editing applied either in parental stem cells or directly in the macrophages by lentiviral vector transduction. Conclusions We show that porcine PSCdMs exhibit key macrophage characteristics, including infection by a range of commercially relevant pig pathogens. In addition, genetic engineering of PSCs and PSCdMs affords new opportunities for functional analysis of macrophage biology in an important livestock species. PSCs and differentiated derivatives should therefore represent a useful and ethical experimental platform to investigate the genetic and molecular basis of host-pathogen interactions in pigs, and also have wider applications in livestock.

Stimulation in one sensory modality can affect perception in a separate modality, resulting in diverse effects including illusions in humans. This can also result in cross-modal facilitation, a process where sensory performance in one modality is improved by stimulation in another modality. For instance, a simple sound can improve performance in a visual task in both humans and cats. However, the range of contexts and underlying mechanisms that evoke such facilitation effects remain poorly understood. Here, we demonstrated cross-modal stimulation in wild-caught túngara frogs, a species with well-studied acoustic preferences in females. We first identified that a combined visual and seismic cue (vocal sac movement and water ripple) was behaviourally relevant for females choosing between two courtship calls in a phonotaxis assay. We then found that this combined cross-modal stimulus rescued a species-typical acoustic preference in the presence of background noise that otherwise abolished the preference. These results highlight how cross-modal stimulation can prime attention in receivers to improve performance during decision-making. With this, we provide the foundation for future work uncovering the processes and conditions that promote cross-modal facilitation effects.

Females of many species choose mates using multiple sensory modalities. Multimodal noise may arise, however, in dense aggregations of animals communicating via multiple sensory modalities. Some evidence suggests multimodal signals may not always improve receiver decision-making performance. When sensory systems process input from multimodal signal sources, multimodal noise may arise and potentially complicate decision-making due to the demands on cognitive integration tasks. We tested female túngara frog, Physalaemus (=Engystomops) pustulosus, responses to male mating signals in noise from multiple sensory modalities (acoustic and visual). Noise treatments were partitioned into three categories: acoustic, visual, and multimodal. We used natural calls from conspecifics and heterospecifics for acoustic noise. Robotic frogs were employed as either visual signal components (synchronous vocal sac inflation with call) or visual noise (asynchronous vocal sac inflation with call). Females expressed a preference for the typically more attractive call in the presence of unimodal noise. However, during multimodal signal and noise treatments (robofrogs employed with background noise), females failed to express a preference for the typically attractive call in the presence of conspecific chorus noise. We found that social context and temporal synchrony of multimodal signaling components are important for multimodal communication. Our results demonstrate that multimodal signals have the potential to increase the complexity of the sensory scene and reduce the efficacy of female decision making.

Noise is a common problem in animal communication. We know little, however, about how animals communicate in the presence of noise using multimodal signals. Multimodal signals are hypothesised to be favoured by evolution because they increase the efficacy of detection and discrimination in noisy environments. We tested the hypothesis that female túngara frogs' responses to attractive male advertisement calls are improved in noise when a visual signal component is added to the available choices. We tested this at two levels of decision complexity (two and three choices). In a two-choice test, the presence of noise did not reduce female preferences for attractive calls. The visual component of a calling male, associated with an unattractive call, also did not reduce preference for attractive calls in the absence of noise. In the presence of noise, however, females were more likely to choose an unattractive call coupled with the visual component. In three-choice tests, the presence of noise alone reduced female responses to attractive calls and this was not strongly affected by the presence or absence of visual components. The responses in these experiments fail to support the multimodal signal efficacy hypothesis. Instead, the data suggest that audio-visual perception and cognitive processing, related to mate choice decisions, are dependent on the complexity of the sensory scene.

Communication systems often include a variety of components, including those that span modalities, which may facilitate detection and decision-making. For example, female túngara frogs and fringe-lipped bats generally rely on acoustic mating signals to find male túngara frogs in a mating or foraging context, respectively. However, two additional cues (vocal sac inflation and water ripples) can enhance detection and choice behavior. To date, we do not know the natural variation and covariation of these three components. To address this, we made detailed recordings of calling males, including call amplitude, vocal sac volume, and water ripple height, in 54 frogs (2430 calls). We found that all three measures correlated, with the strongest association between the vocal sac volume and call amplitude. We also found that multimodal models predicted the calling males' mass better than unimodal models. These results demonstrate how multimodal components of a communication system relate to each other and provide an important foundation for future studies on how receivers integrate and compare complex displays.

As species change through evolutionary time, the neurological and morphological structures that underlie behavioral systems typically remain coordinated. This is especially important for communication systems, in which these structures must remain coordinated both within and between senders and receivers for successful information transfer. The acoustic communication of anurans (“frogs”) offers an excellent system to ask when and how such coordination is maintained, and to allow researchers to dissociate allometric effects from independent correlated evolution. Anurans constitute one of the most speciose groups of vocalizing vertebrates, and females typically rely on vocalizations to localize males for reproduction. Here, we compile and compare data on various aspects of auditory morphology, hearing sensitivity, and call-dominant frequency across 81 species of anurans. We find robust, phylogenetically independent scaling effects of body size for all features measured. Furthermore, after accounting for body size, we find preliminary evidence that morphological evolution beyond allometry can correlate with hearing sensitivity and dominant frequency. These data provide foundational results regarding constraints imposed by body size on communication systems and motivate further data collection and analysis using comparative approaches across the numerous anuran species.

To advance our understanding of adaptation to temporally varying selection pressures, we identified signatures of seasonal adaptation occurring in parallel among Drosophila melanogaster populations. Specifically, we estimated allele frequencies genome-wide from flies sampled early and late in the growing season from 20 widely dispersed populations. We identified parallel seasonal allele frequency shifts across North America and Europe, demonstrating that seasonal adaptation is a general phenomenon of temperate fly populations. Seasonally fluctuating polymorphisms are enriched in large chromosomal inversions, and we find a broad concordance between seasonal and spatial allele frequency change. The direction of allele frequency change at seasonally variable polymorphisms can be predicted by weather conditions in the weeks prior to sampling, linking the environment and the genomic response to selection. Our results suggest that fluctuating selection is an important evolutionary force affecting patterns of genetic variation in Drosophila.

Many animals acoustically communicate in large aggregations, producing biotic soundscapes. In turn, these natural soundscapes can influence the efficacy of animal communication, yet little is known about how variation in soundscape interferes with animals that communicate acoustically. We quantified this variation by analyzing natural soundscapes with the mid-frequency cover index and by measuring the frequency ranges and call rates of the most common acoustically communicating species. We then tested female mate choice in the túngara frog (Physalaemus pustulosus) in varying types of background chorus noise. We broadcast two natural túngara frog calls as a stimulus and altered the densities (duty cycles) of natural calls from conspecifics and heterospecifics to form the different types of chorus noise. During both conspecific and heterospecific chorus noise treatments, females demonstrated similar preferences for advertisement calls at low and mid noise densities but failed to express a preference in the presence of high noise density. Our data also suggest that nights with high densities of chorus noise from conspecifics and heterospecifics are common in some breeding ponds, and on nights with high noise density, the soundscape plays an important role diminishing the accuracy of female decision-making.

Individuals produce advertisement signals with intended purposes and targets. However, these signals can be received by eavesdroppers, who may extract information from them and alter their behavior according to the extracted information. In anuran systems, males congregate at breeding sites to produce advertisement calls to attract receptive females and fend off rival males. Both sexes directly assess these calls in dyadic encounters and make decisions based on the call's characteristics, e.g., frequency. What is unknown is whether bystander males eavesdrop on these same calls to inform their future competitive decisions. Here, we examined whether male green tree frogs (Hyla cinerea) eavesdrop on competing males, assess their competitor's call frequency, and respond accordingly. We exposed males to playbacks of two competing males that varied in call frequencyhigh, average, lowand quantified latency to call, time spent calling, and number of calling bouts. We found that males had reduced latency to call and called more when eavesdropping low-frequency competition, but not average or high-frequency competition. Focal male size also influenced how they responded, with larger males being more responsive than smaller males. Our results indicate that male green tree frogs are capable of eavesdropping on nearby male calls and produce behavioral responses accordingly. Further, it appears males are able to alternate between assessment strategies dependent upon the frequency of the eavesdropped competition. These findings indicate that males not only directly assess an opponent's call in dyadic encounters, but also indirectly through eavesdropping.Significance statementAnimals produce signals with intended purposes and targets, but which can be received by nearby eavesdroppers. Eavesdropped signals can elicit complex phenotypic changes in the eavesdropper, and lead to significant fitness consequences for the signal producer and eavesdropper. Thus, examining whether and to what extent individuals eavesdrop is an important step in understanding the evolution of signal production and response. Here, we examined whether male green tree frogs, H. cinerea, eavesdrop on nearby competing males and base their own calling behaviors on eavesdropped male's calling characteristics. Calling behavior was mediated by focal male body size and eavesdropped male's call frequency. Larger males reduced response latency and called more to nearby low-frequency males, while smaller males reduced response latency to nearby calls of average frequency. These results indicate that males will extract information from their competitive environment through eavesdropping and produce behavioral responses according to the eavesdropped information.

The field of articulatory phonetics is concerned primarily with the question of how speech is realized through movements of body structures. While articulatory phoneticians have often described speech sounds using terms that refer to an inventory of body parts (e.g., “tongue,” “lips,” “velum,” etc.), a core challenge of articulatory phonetics is to understand how such structures function and interact to produce speech sounds. A complete understanding of articulatory phonetics thus requires that we define ways of mapping our descriptive terms onto groupings of nerves and muscles that our brains and bodies can use to produce speech movements. This chapter explores some of the ways articulatory phoneticians describe speech sounds, suggesting that a more fully embodied approach can provide novel insights into speech articulation and can help to understand links between speech and other functions of the human

Amphibian populations are being threatened by human related activities including the spread of the fungal pathogen, Batrachochytrium dendrobatidis (Bd) and urbanization. With growing losses in global amphibian biodiversity, it is essential to document how amphibian populations are responding to rapid environmental changes. While most evolutionary processes, e.g. changes in allelic frequencies, may be too slow to allow adequate response to environmental changes, epigenetic modifications can rapidly translate environmental changes into adaptive phenotypic responses. Epigenetic modifications come in multiple, non-exclusive forms, the most notable being DNA methylation. Here we sought to examine variation in the frequency of DNA methylation among four túngara frog populations distributed across Gamboa, Panama; which vary in both their level of fungal presence/prevalence and urbanization. DNA samples were collected from amplexed (male–female) pairs and frequency of DNA methylation was analyzed using a methylation-sensitive amplified fragment length polymorphism protocol. We found significant variation in DNA methylation among populations, and correlations between Bd infection status and methylation patterns. Urbanization, however, had no influences on the frequency of DNA methylation. These data suggest epigenetic modifications are substantially flexible across fine-scale, environmental gradients and there appears to be possible biologically relevant links between DNA methylation and Bd infection status. Our results provide a basis for future work investigating the causal role epigenetics have in mediating phenotypic response to human-induced, environmental changes.

We have all learned to associate real voices with animated faces since childhood. Researchers use this association, employing virtual faces in audiovisual speech perception tasks. However, we do not know if perceivers treat those virtual faces the same as real faces, or if instead integration of speech cues from new virtual faces must be learned at the time of contact. We test this possibility using speech information that perceivers have never had a chance to associate with simulated faces – aerotactile somatosensation. With human faces, silent bilabial articulations (“ba” and “pa”), accompanied by synchronous cutaneous airflow, shift perceptual bias towards “pa”. If visual-tactile integration is unaffected by the visual stimuli’s ecological origin, results with virtual faces should be similar. Contra previous reports [8], our results show perceivers do treat computer-generated faces and human faces in a similar fashion visually aligned cutaneous airflow shifts perceptual bias towards “pa” equally well with virtual and real faces.