Integrative Organismal Biology

Specialized suction discs are functional innovations that enable fishes to attach to diverse surfaces and resist hydrodynamic forces. The adhesive discs of clingfishes (Gobiesocidae) vary in size and shape, but it is unclear how ecological factors have influenced their morphological evolution. Here we analyzed the disc and skull shape of 74 clingfish species using micro-CT scanning and 3D geometric morphometrics to investigate the role of habitat and substrate use on patterns of diversification. We also present novel comparisons of adhesive performance for 10 clingfish species. Clingfish interface directly with their environment using their adhesive discs, but we found that the disc and skulls are evolutionary integrated and share similar responses to habitat. Transitions from coastal habitats to coral reefs promoted elevated rates of evolution and morphological disparity across the body, whereas transitions to freshwater did not. Concurrently, repeated shifts from living on hard substrates to softer substrates (i.e., macroalgae and seagrass) were associated with more constrained disc shapes, convergent morphologies, and differences in adhesive performance. We propose that habitat and substrate use make complementary contributions to skeletal diversification in clingfishes, but the adhesive system requires further investigation to disentangle the complex interactions between form and function.

Shorebirds provide an excellent model for investigating the relationship between bill morphology and food acquisition. Food acquisition comprises three successive behavioural stages: foraging (locomotion and prey capture), feeding (food handling and transport), and swallowing. During food transport, these birds use two non-lingual mechanisms, surface-tension transport (ST) and ballistic transport (BT), whose characteristics depends on the kinematics of head and beak movements and the physical properties of the food. We investigated the behavioural flexibility of these transport mechanisms in captive and free-ranging individuals of two species with contrasting beak morphologies: the Pied avocet (Recurvirostra avosetta) and the Black-winged stilt (Himantopus himantopus). Although these species have beaks of a similar size, avocets are distinguished by their upward-curved beaks, whereas stilts have a rather straight beak. We examined the effects of food water content and the presence or absence of water in the beak on food transport by quantifying maximum gape, maximum head displacement, and maximum head velocity. Transport kinematics were jointly influenced by food properties and water availability in the beak. Moist food improved transport performance in both species, whereas dry food required compensatory increases in gape amplitude and head movements, demonstrating that neither ST nor BT constitutes a fixed behavioural sequence. Species also differed consistently in their transport strategies: Black-winged stilts relied on slower, larger-amplitude head movements, whereas Pied Avocets exhibited faster, more precise movements, particularly when water was present. These findings demonstrate that ST and BT share common biomechanical foundations while being governed by rapid kinematic adjustments to changing environmental conditions that probably correspond to flexible motor control. This behavioural flexibility in food transport is therefore likely to enhance feeding performance and ecological resilience in the heterogeneous habitats occupied by shorebirds, suggesting that context-dependent modulation of transport behaviour represents an important adaptive feature of these shorebirds.

When food resources provide reward while simultaneously imposing risk, animals can regulate their behaviour to continue exploiting them while reducing associated costs. Angling presents such a feeding situation, in which obtaining bait can result in hooking and capture. Although fish become less vulnerable to angling following capture experience, the behavioural changes underlying this reduced vulnerability remain poorly understood. Here, we used fine-scale video observations to examine feeding behaviour of red sea bream (Pagrus major) toward angling gear before and after capture experience. Naive fish rapidly swallowed the bait and were subsequently captured, whereas capture-experienced fish continued to approach and feed on the gear but shifted from swallowing to biting. This feeding-mode shift may allow fish to maintain exploitation of the bait while reducing capture risk. Our observations provide a behavioural explanation for reduced angling vulnerability and suggest that direct experience with capture risk can alter how fish exploit a rewarding food resource.

The ability of prey to eavesdrop on predator vocalizations is expected to increase survival by reducing detection and capture. Unfortunately, most research has been conducted in vertebrates, and little is known about this ability in invertebrates. We measured latency to emerge, overall activity, and shelter visits in wild-caught fall field crickets (Gryllus pennsylvanicus) in response to acoustic playback. Stimuli included multiple predator vocalizations, non-predator vocalizations, white noise, and a control. We predicted that crickets would reduce activity, spend more time in shelter, and freeze in response to stimuli representing greater risk. Contrary to our predictions, crickets traveled greater distances, spent more time moving, and spent less time in shelter in response to predator vocalizations versus controls. We did not, however, find clear differences in responses between predator vocalizations and other treatments. Our results suggest that crickets may not differentiate between the vocalizations of predators, non-predators, and other abrupt sounds. Consequently, eavesdropping may not be a viable method of assessing predation risk for this species and its general use remains unclear.

An animal's musculature must support its specific biomechanical needs, so muscle morphology and volume allocation may adapt when locomotor strategies diversify. We examined muscle size and morphology in two sister families of bombycoid moths, wild silkmoths (Saturniidae) and hawkmoths (Sphingidae), that have diverged in wingbeat frequency, wing morphology, and behavior. Although both families rely on the same muscles to power and steer flight, they may distribute muscle volume differently to prioritize distinct functions. We hypothesized that flight power muscle proportions are larger in hawkmoths and increase with wingbeat frequency, helping meet inertial power demands of high-frequency maneuverable flight. We also hypothesized that some individual muscles diverge in proportional volume and area to support distinct wing control strategies. To test our hypotheses, we took CT scans of twenty bombycoid species and quantified volumes and geometries of six flight muscle pairs. As expected, flight power muscle proportions positively correlate with wingbeat frequency and are generally greater in hawkmoths. Two of three steering muscles diverge substantially in relative volume and area between families. Most muscles exhibit greater length in silkmoths and greater cross-sectional area in hawkmoths. Finally, the dorsal oblique(DO) muscle diverges exceptionally in size and morphology, being highly developed in hawkmoths and smaller or absent in silkmoths. This unexpected difference supports the DO having an underappreciated role in flight control, possibly via shaping indirect strain propagation in the elastic thorax. We show that muscle volume distribution parallels bombycoids' divergent flight strategies, demonstrating how muscle allocation can adapt for specialized functional goals.

Gliding enables mammals to forage and escape from predators by moving between discontinuous forests. Its benefits depend not only on glide distance but also on the ability to decelerate and land safely. This study developed a theoretical framework linking glide distance, gliding velocity, aerodynamic braking, body mass, and braking distance. Twenty-two representative distance-velocity observations from eight studies and five species were compiled. Among distance-velocity models, log-distance and saturated with V0 models received nearly equivalent support. Both models predicted increasing velocity with glide distance, with the rate of increase declining at longer distances. For a 1 kg animal undergoing a 60% reduction in velocity, predicted kinetic energy remaining immediately before contact increased from 4.80-5.14J at 20 m to 8.13-8.90 J at 80 m. This velocity reduction corresponded to a dissipation of 84% of approach kinetic energy before contact. Over a braking distance of 1 m, the required mean deceleration increased from 2.57-2.75 g at 20 m to 4.35-4.77 g at 80 m. At 80 m, shortening the braking distance from 4 to 0.5 m increased the required deceleration from 1.09-1.19 to 8.70-9.53 g. These results indicate that the absolute energetic and deceleration requirements of landing increase with glide distance, even when velocity is close to an asymptote. These results provide a quantitative basis for considering aerodynamic braking and landing requirements alongside conventional measures of glide performance. Summary statementModels quantify how glide distance, aerodynamic braking, and braking distance affect pre-contact kinetic energy and deceleration requirements in gliding mammals.

This study investigates the impact temperature-induced vertebral anomalies have on the C-start escape response of Astyanax mexicanus, a model species in evolutionary developmental biology. Employing three temperature treatments to induce varying degrees of skeletal anomalies, we assessed their effects on key swimming performance metrics including, C-start time, curvature coefficient, head displacement distance, and displacement velocity. Through the use of linear mixed models and generalized linear mixed models, our results reveal that specific anomalies such as vertebral fusions and anomalous haemal and neural spines affected the curving ability of C-start escape responses. However, these did not negatively impact other performance parameters, with velocity, distance, and response time showing no significant impacts from any anomaly types, when assessed individually. This suggests a complex interplay between structural deformities and compensatory physiological mechanisms that maintain functional performance. Other variables measured had a stronger and significant impact on swimming performance, including standard length, vertebral number, and temperature treatment, which influenced escape speed, curving ability, and overall locomotor performance. Our findings challenge conventional perceptions about the debilitating impact of vertebral anomalies, indicating that many affected fish can still effectively perform escape maneuvers critical for survival.

Cruising speed is a key factor affecting prey-search efficiency and migration range in continuously swimming animals. Tunas and lamnid sharks (e.g., white sharks) have convergently evolved traits for high-speed cruising, including the ability to maintain slow-twitch, aerobic red muscle (RM) warmer than ambient water, known as RM endothermy. Despite their well-known high cruising speeds, kinematic features underlying their elevated speeds remain unclear. Swim speed is the product of tailbeat frequency (TBF; Hz) and stride length (SL, the absolute distance traveled per tailbeat; m). RM endothermy is expected to elevate TBF by enhancing muscle contraction performance. Furthermore, within RM-endothermic fishes, tunas and lamnid sharks may exhibit distinct kinematic features because of differences in caudal fin morphology and tailbeat amplitude. Here, we compiled kinematic parameters from 20 fish species, including five RM-endothermic species, measured in the wild using animal-borne sensors. Comparative analyses showed that, for a given body mass and water temperature, RM-endothermic fishes exhibited 1.9 times higher cruising speed and TBF than ectothermic fishes, while SL remained similar. Within RM-endothermic fishes, tunas exhibited 2.3 times higher TBF than similar-sized lamnid sharks, whereas lamnid sharks showed 1.7 times longer SL than similar-sized tunas. These results indicate that RM endothermy is generally associated with higher TBF, while significant kinematic differences remain between tunas and lamnid sharks. This divergence may be partly explained by the greater caudal fin area and tailbeat amplitude in lamnid sharks. It may also reflect contrasting skeletal types of teleosts and elasmobranchs, which potentially influence body stiffness and swimming kinematics.

Sexual dimorphism is a well-studied phenomenon in which males and females of a species exhibit different phenotypic characteristics. However, the degree of sexual dimorphism within a species may vary among populations and may be influenced by geographic factors such as seasonality, prey availability, and population density. In this study, we examined the effects of sexual dimorphism and geographic region on the craniomandibular morphology of bobcats (Lynx rufus) in Washington State. Using digital calipers, we measured 13 craniomandibular traits from 35 males and 42 females. We found that the degree of skull dimorphism does not differ between the eastern and western side of the Cascade Mountains. However, we found significant effects of sex and geographic region on craniomandibular morphology. Across the geographic regions, males were significantly larger than females in most craniomandibular traits, excluding postorbital constriction breadth (POC). After correcting for expected size differences, only condylobasal length (CBL) was disproportionately longer in males, indicating subtle shape dimorphism within this trait, that is largely unexplained by allometry. Mechanical advantage (MA) of the temporalis and masseter muscles did not differ between the sexes at the canine or the molar bite points. Comparisons between populations east and west of the Cascade Range revealed that while sex remains the primary indicator of overall size, there is a small regional effect on size as western bobcats are slightly larger than eastern bobcats. These regional differences highlight how environmental pressures including local prey availability and severe winter conditions may influence functional skull design. Future ecological and behavioral studies are needed to further clarify how these localized environmental drivers shape phenotypic variation across populations.

Multifunctional anatomical structures must accommodate potentially competing performance demands, leading to functional tradeoffs. The mammal rostrum (snout) is involved in multiple functions, including feeding, respiratory conditioning, olfaction, and, in most bats, echolocation. Therefore, it serves as an ideal system to investigate whether and how functional tradeoffs promote or constrain morphological diversification. While the external morphology of the rostrum has been linked with mechanical demands of feeding in bats, the factors and processes influencing its internal morphology, including the turbinals in the nasal cavity, remain comparatively unexplored. Here we investigate how internal rostral morphology reflects specialization and tradeoffs among respiratory air conditioning, echolocation, and olfaction by quantifying turbinal morphological diversity in a phylogenetically and ecologically diverse sample of bat species. We find that echolocation mode predominantly influences both the ethmoturbinal complex (ETc) and maxilloturbinal (MT) systems, with nasal-emitting bats having reduced and less complex turbinals compared to oral and non-laryngeally echolocating bat species. MT complexity and surface area are not associated with bioclimatic variables representing thermal variability and aridity. However, the ETc exhibits higher complexity in plantivorous bats than faunivorous bats, suggesting higher olfactory reliance. While MT morphological disparity was predominantly explained by echolocation mode, ETc morphological disparity was associated with both echolocation mode and diet, suggesting potentially opposing demands favoring turbinal reduction for sound transmission and elaboration for olfaction. Among nasal emitters, which must accommodate both demands within the nasal cavity, ETc complexity exhibited higher relative disparity compared to MT complexity. These patterns suggest that competing functional demands may promote or maintain morphological diversity, possibly through ecological specialization and/or many-to-one mapping between morphology and performance.

As foraging increases exposure to predators, individuals must allocate time to vigilance in addition to food acquisition. Although those behaviors are assumed to be mutually exclusive, this is often hard to disentangle, given that multiple sensory modalities are often applied. However, when foraging, animals often have one sensory modality more impaired (e.g. vision), than others (e.g. auditory); thus, a level of compromise on vigilance capabilities is assumed while foraging. Here, we test the idea that when a food item requires manipulation, thus reducing vigilance capabilities, social foraging should promote shared costs of vigilance. We conducted video recordings to investigate whether there is a trade-off between foraging and vigilance behavior in red-legged seriemas (Cariama cristata) when foraging in pairs. We studied the social foraging behavioral patterns while seriemas were feeding on macauba palm (Acrocomia aculeata) fruit, which requires a series of throwing maneuvers to crack the hard outer shell. Our results showed that within a pair recording one individual was observed to spend more time vigilant than their partner, while the other individual spent more time throwing fruit than their partner. However, we found no evidence for synchronization or coordination of vigilance and foraging between individuals within a feeding bout. These results provide insight into the foraging ecology of red-legged seriemas and the trade-off between foraging and vigilance in social contexts.

1. Adaptive behavioural strategies require animals to simultaneously navigate social and ecological domains across multiple spatial and temporal scales. Although drones and computer vision have recently transformed the study of wild animal societies, many nocturnal species and those occupying structurally complex habitats remain inaccessible to these approaches, limiting our understanding of behaviour in natural settings. 2. We aimed to determine how behavioural strategies are organised across nested spatial and temporal scales within a wild communication network. 3. We used three-dimensional acoustic localisation and source separation to track individual male Hyperolius sp. A, a nocturnal African reed frog, within a natural rainforest chorus and quantify patterns of site fidelity, movement, spatial organisation, and call-timing interactions. 4. Males exhibited significant site fidelity across nights, while chorus spatial structure varied with local caller density. Within nights, individuals followed a stereotyped behavioural sequence, descending from elevated arboreal refugia before settling into lower calling positions near breeding sites. At finer temporal scales, call-timing interactions varied according to both local competitor density and the proximity of neighbouring rivals. 5. These findings demonstrate that behavioural strategies emerge across nested spatial and temporal scales and that long-term spatial positioning, short-term movement decisions, and moment-to-moment signalling interactions are tightly linked within natural communication networks. More broadly, acoustic localisation provides a powerful framework for studying behaviour in species and habitats that remain difficult to observe using conventional approaches.

Developing robust methods to quantify how animals allocate time across behaviours is essential for understanding energy use, habitat requirements, and responses to environmental change. For cryptic, semi-aquatic mammals such as the platypus, direct observation is difficult, creating a reliance on remote biologging approaches that can reliably infer behaviour in the wild. However, aquatic environments can both smooth acceleration signals through hydrodynamic damping and introduce noise from water movement, turbulence, and drag, potentially obscuring behavioural differences of similar magnitudes. We tested whether progressively incorporating biomechanical and frequency-domain (FFT-derived) predictors improved behavioural classification in hydrodynamically challenging aquatic environments. Tri-axial accelerometers were deployed on four ex situ platypuses, with synchronised video observations used to validate behaviour. From the acceleration data, we derived three predictor classes of increasing complexity: summary statistics describing activity level, engineered biomechanical variables capturing posture and body orientation, and FFT-derived features describing movement rhythm. These predictors were progressively incorporated into Random Forest models to classify five behaviours: burrow resting, surface resting, grooming, travelling/foraging, and diving. Model performance improved with increasing predictor complexity, although gains were behaviour specific. FFT-derived features substantially improved classification of rhythmic behaviours such as diving and foraging, while engineered biomechanical predictors improved grooming detection. In contrast, resting behaviours, particularly surface resting, showed little improvement. Overall accuracy increased from [~]75% to [~]88% when frequency-domain features were included. Misclassification was greatest among behaviours with overlapping or low-amplitude signals, and cross-individual validation revealed reduced model generalisability, indicating that individual variation in movement patterns constrained transferability. Incorporating frequency-domain features substantially improved behavioural classification in platypuses, particularly for rhythmic behaviours such as diving and foraging. This study provides the first validated accelerometry-based behavioural classification framework for the species and highlights the importance of matching predictor selection to behavioural mechanics. More broadly, the approach offers a transferable framework for aquatic and semi-aquatic taxa.

Humans tend to walk at speeds that minimize energy expenditure and time duration. While most walking experiments examine individuals in relative isolation, everyday locomotion frequently occurs in social contexts. We investigated whether walking speed is modulated in response to a social interaction during a cooperative task. Participants completed 96 randomized walking trials where they approached and retrieved boxes varying in distance (2.5-10 m) and mass (0-6.8 kg). Boxes either rested on the ground or were handed off by an experimenter, signaling prosocial effort benefitting the participant. Approach speed was measured with inertial measurement units placed at the feet and fit to a saturating exponential function of walking distance using a nonlinear mixed-effects regression model. Based on the energy-time optimization framework, we hypothesized that participants would approach more quickly when larger boxes were held at farther distances, to reduce energy and time costs of the experimenter, despite exerting more effort themselves. Participants walked 8.5% faster (1.29 m s-1 versus 1.19 m s-1; p = 3.85 x 10-6) when the largest box was held out by the experimenter versus left on the ground. However, the manner in which the box was held had no influence on approach speeds (p = 0.31). Exploratory analyses identified modest trends between experiment responses and individual characteristics, but none reached statistical significance. The findings suggest that locomotor decisions reflect not only an individuals own energy and time costs but also the perceived costs borne by others. This study demonstrates that social context can meaningfully influence walking behavior. Summary StatementHumans walk more quickly in response to the perceived energy and time others spend helping them, even at the cost of exerting more energy themselves.

Animals face a fundamental trade-off between food-related competition and predation risk in how they allocate time and behavior. Nocturnal mammals offer a particularly tractable system for testing this trade-off because moonlight creates a natural, quantifiable gradient in both predation risk and the visibility needed for safe movement. We used minute-by-minute focal observations of nine kinkajous (Potos flavus; 4 female, 5 male) in Panama to test how fruit abundance and moonlight predicted nocturnal activity budgets (percent time traveling, feeding, resting) and nightly travel distance. Beta-family generalized linear mixed models and a linear mixed model of log travel distance showed that fruit abundance was positively associated with percent time traveling and with nightly travel distance, and negatively associated with percent time feeding: kinkajous traveled more and fed less per hour when fruit was abundant, consistent with movement between many nearby productive trees rather than prolonged feeding at a few. Males traveled less as moonlight increased, while females traveled more. Rainfall had no independent effect. We interpret the sex-reversed moonlight response as evidence that moonlight elevates predation risk for males while facilitating movement for the more food-limited females of this frugivorous carnivore.

Metabolic rate governs the pace of life through the allocation and expenditure of energy, yet metabolic rate data are unavailable for most species. External morphological traits, such as gill slits, have been used to infer respiratory physiology and metabolism in sharks, yet, whether that pattern is generalizable across rays is unknown. In rays (Batoidea), spiracle length seems adaptive to oxygen uptake capacity, environmental oxygen availability, and activity level. Thus, we evaluate the allometry of external respiratory morphology (spiracles and gill slits) in rays using 550 museum specimens representing 88 species. We test whether spiracle length and gill slit height grow isometrically with body size (total length or disc width) within and across species. Overall, we find spiracles and gill slits are morphologically diverse, varying widely in shape within and between families. Spiracle length and gill slit height scale isometrically and are a consistent proportion of body size, despite such high morphological variability. While that proportion is consistent through ontogeny, each species proportion differs (e.g. gill slit height ranges from 0.93% of total length in Spotted Legskate to 2.93% in Pacific Electric Ray). Of the three swimming styles, pectoral oscillating species (Mobulid and Eagle Rays) may have steeper scaling of spiracle length with gill slit height - consistent with greater respiratory capacity in species with more active lifestyles. Our findings are consistent with literature hypothesizing external respiratory morphology (spiracles and gill slits) is tightly related to metabolic rate and activity level, and demonstrate the value of natural history collections in ecological research. Importantly, the isometry of both respiratory traits means that a single measure of any individual, irrespective of size, will be representative of the species thus providing a basis for expanding our understanding of metabolic morphology, enabling future work on life history, extinction risk, and ecological function.

Koalas (Phascolarctos cinereus) are marsupials highly specialized for arboreal life and are considered a representative example of convergent evolution with primates, which are also adapted to arboreal habitats. The forelimb digital flexors play a crucial role in arboreal locomotion by flexing the digits and enabling the claws or fingers to engage with tree trunks and branches. Although the morphology and function of the digital flexors have been extensively examined in primates, anatomical information on koala digital flexors has not been sufficiently updated in modern anatomical studies. In this study, we examined the digital flexor muscles and their innervation in four forelimbs from three koalas to clarify both shared and distinctive structural features among arboreal mammals. The flexor digitorum profundus (FDP) was markedly developed, formed a U-shaped muscle belly, and received dual innervation from the median and ulnar nerves. In contrast, the flexor digitorum superficialis (FDS) was small, arose from the superficial surface of the FDP, was partially enclosed by the FDP muscle belly, and gave rise to extremely slender tendons inserting into digits II-V. The branch supplying the FDS arose from the main trunk of the median nerve near the carpal region, ascended proximally in a recurrent course, and terminated within the FDS. This unusual course may be consistent with a morphogenetic process underlying FDS formation. These findings indicate that the koala digital flexors are characterized by a markedly developed FDP, a relatively reduced FDS, and a distinctive innervation pattern of the FDS. Thus, even among mammals adapted to arboreal life, koalas and primates appear to have achieved arboreal adaptation through different morphological specializations within the forearm digital flexor system.

Tessellated calcified cartilage (TCC) is a hallmark of the chondrichthyan skeleton, yet its development early in ontogeny across the four major groups (batoids, galeomorphs, squalomorphs, and holocephalans) remains poorly understood. Specialised traits of TCC, such as multi-layered TCC and internal mineralised trabeculae, typically develop in response to feeding mechanics. In this study, we evaluated TCC morphology in the jaws of 12 representative taxa to observe its structure at an early ontogenetic stage to determine whether these specialised features had yet developed. Batoids consistently exhibited well-developed, homogeneous, polygonal tesserae early in ontogeny regardless of jaw morphology or feeding habit. In contrast, galeomorphs displayed high morphological heterogeneity. Notably, we document the first report of an extensive internal trabecular network in a non-batoid elasmobranch, observed in Ginglymostoma cirratum, which may serve to resist the mechanical pressures of specialised suction feeding. Furthermore, we identified voussoir tesserae in galeomorphs for the first time, extending their documented presence across all elasmobranch groups, where they display an inverted aspect ratio (wider than tall) compared to mature forms. The durophagous Mustelus mustelus exhibited surprisingly poor TCC development despite being a durophagous feeder, pointing to a pronounced ontogenetic lag. In Squatina oculata, TCC was characterised by large and thick tesserae and extensive fused tesseral regions which may relate to its explosive ambush predation mode, whereas the holocephalan Chimaera exhibited a poorly mineralized, mesh-like structure without resolvable discrete tesserae or trabeculae-matching findings from previous studies. Across all specimens, multi-layered TCC was absent, confirming that multi-layering develops later in ontogeny. These results demonstrate that generalised models of TCC development based on one group or a few taxa fail to capture the broader diversity of TCC morphology. It also opens up many exciting avenues for further study, and forms the basis for comparisons with fossil chondrichthyans, to investigate the evolution of TCC.

Honey bees can swim on the water surface toward dark regions, a behavior known as scototaxis that may facilitate escape from water. Although this behavior has been reported in both honey bees and solitary bees, variation among honey bee species remains poorly understood. We compared scototaxis during swimming in four honey bee species representing two nesting types: open-nesting (Apis florea and A. dorsata) and cavity-nesting (A. cerana and A. mellifera). Individual bees were released into a water-filled arena containing a dark sector, and their landing angles were recorded. All species exhibited significant orientation toward the dark sector. However, open-nesting species showed significantly stronger orientation than cavity-nesting species. No significant differences were detected between replicate colonies within species or between species within the same nesting type, whereas differences between nesting types were highly significant. Hierarchical clustering based on orientation strength placed Osmia, a solitary cavity-nesting bee from our previous study, in the same behavioral cluster as the two open-nesting Apis species rather than the cavity-nesting honey bees. We also measured swimming duration, distance, and velocity, but found no consistent differences between nesting types. These results demonstrate substantial interspecific variation in swimming scototaxis. The behavioral clustering is consistent with the hypothesis that strong scototaxis represents an ancestral trait that has been reduced in the derived A. cerana/A. mellifera lineage.

Female-female competition is increasingly recognized as a key driver of female ornamentation, including birdsong, which often functions in intrasexual competition. However, the specific resources females use elaborate traits to compete for remain unclear. In addition, few studies have simultaneously investigated the use of multiple competitive traits in females, despite growing independent interest in these traits (e.g., female song and aggression). We investigated the competitive contexts of female song, aggression and calling behavior in northern house wrens (Troglodytes aedon) to determine which resources females compete for across the breeding season. We simulated conspecific territorial intrusions using female song at three breeding stages representing different contexts: arrival (mate and territory acquisition), nest building (nest site and breeding status defense), and egg laying (brood defense). We tested whether female song and physical aggression varied as reproductive resources shifted across the breeding cycle. Females were significantly more aggressive during nest building, showing 5.8 times greater odds of a higher-intensity aggressive response during nest building compared to arrival. Female song output was similar across early stages but declined during egg laying, though this was not statistically significant after correction for multiple comparisons and individuals varied substantially in overall singing propensity. Non-song vocalizations varied by call type and breeding stage. Calls associated with aggression occurred most frequently during nest building, consistent with peak physical aggression responses. Together, these results identify nest building as the stage of highest female aggression, consistent with heightened competition over nest cavities and associated breeding status in this cavity-nesting species. In contrast, female song occurred across all stages and appears to function in multiple competitive contexts. This study provides evidence for context and mode-specific female signaling in a temperate songbird and highlights that females strategically use aggression, calls, and song to mediate social conflict across breeding contexts.