Integrative And Comparative Biology

The honey bee colony (Apis mellifera) acts as a superorganism, with a dual immune system that operates at the individual and social level. However, the linkages between immune mechanisms across the two levels remain poorly understood, despite the relevance for developing effective breeding strategies to improve honey bee disease resistance. Hygienic behavior involving the removal of unhealthy brood is a key component of honey bee social immunity and is highly effective at limiting parasites and pathogens in the colony. While this form of hygienic behavior can reduce brood diseases, parasites infecting adult bees primarily, such as Nosema ceranae, are not directly impacted by the behavior. However, when using the Unhealthy Brood Odor (UBeeO) assay to quantify hygienic behavior performance, hygienic colonies have been shown to maintain lower Nosema spp. loads over time and overall compared to non-hygienic colonies. To investigate the mechanisms driving reduced Nosema spp. in hygienic colonies, we conducted a series of field and lab experiments to test the innate immune performance of individual bees. We evaluated several factors across hygienic and non-hygienic bees including (1) differences in N. ceranae infection levels, (2) survival probability, (3) Vitellogenin and Hymenoptaecin gene expression, and (4) amount of N. ceranae inoculant consumed. We found that hygienic bees consumed less of the inoculant, exhibited upregulated Vitellogenin gene expression at peak N. ceranae infection, showed a positive relationship between Hymenoptaecin gene expression and N. ceranae infection levels, and had greater survivability when infected with N. ceranae, compared to non-hygienic bees. Here, we present new findings that link colony hygienic behavior performance to individual-level resistance and tolerance mechanisms in response to N. ceranae, suggesting broader implications for the success of selective breeding programs targeting hygienic traits.

Many benthic marine invertebrates disperse by releasing microscopic larvae carried by ocean currents to new sites, where they must settle into appropriate habitats and metamorphose to recruit. Species whose larvae settle in response to water-borne chemical cues live in topographically complex habitats. To study whether sinking in response to dissolved cues affects retention of larvae within complex habitats exposed to ambient water flow moving faster than larvae sink, we used the reef-dwelling sea slug, Phestilla sibogae, whose competent larvae stop swimming and sink in response to dissolved cue from their prey coral, Porites compressa. We conducted field experiments where dye-labelled water, neutrally buoyant particles, and larval mimics (particles that sank at the velocity of larvae of P. sibogae) were released together upstream of reefs of branching corals to determine if larval sinking in water above and within a reef affects larval retention within the reef. Wave-driven water flow measured above a reef in the field had instantaneous velocities peaking at 0.3 m s-1, driving slow net advection of water shoreward at [~]0.02 m s-1. Much slower wave-driven flow moved through the interstices within the reef. In this field flow, sinking by larval mimics caused their retention within a reef after dye-labelled water and neutrally buoyant particles had left. Such retention of sinking larvae within topographically complex benthic communities enhances successful recruitment by exposing larvae to high concentrations of cue for long periods, allowing them time to sink to surfaces, adhere, and undergo metamorphosis.

Finding resources for the colony is one of the most difficult and risky tasks for a social insect worker. A worker on a foraging trip can face a number of challenges, including interference from other individuals, her own errors, and environmental disturbances. Collectively, colonies may use a variety of strategies to minimize the impact of such perturbations on the foraging process. Here, we investigated how individual Solenopsis xyloni ant workers react to perturbation of an established pheromone trail. We trained foragers from colonies in the field to either a low or high concentration sucrose solution in a feeder on a T-maze setup, then replaced a section of floor covering, removing a section of the pheromone trail previously laid. We found that while ants made correct choices on the T-maze when the trail was intact, their choices did not differ from chance when the trail was absent, indicating strong reliance on a pheromone trail (and not, for example, memory) to return to the resource. Moreover, when the trail was absent, we found that a majority of ants abandoned the resource, and that even the ants that were able to reach the resource did not repair the perturbed trail. However, with a high-quality resource, more ants persisted in attempting to reach it (instead of abandoning). We interpret these responses in the framework of robustness mechanisms discussed in systems biology. Our study thus links individual and collective responses to perturbations, and provides an empirical example of how information use interacts with system robustness. Statements and declarationsThe authors have no competing interests to declare that are relevant to the content of this article.

Nocturnal migration is a remarkable phenomenon observed in many insect species, including moths. Migratory moths are capable of maintaining precise directional orientation during migration, as demonstrated in both laboratory and field studies, suggesting that they use multiple environmental cues for orientation and navigation. Recent studies on Australian Bogong moths revealed that these animals can use stellar cues and likely the geomagnetic field (in conjunction with local visual cues) to select and maintain population-specific migratory direction. However, the underlying orientation mechanisms used by most other migratory moths are still largely unresolved. Further, it remains unclear whether migratory moths can adjust their orientation using Earths magnetic field parameters for determining their position relative to the goal (i.e. location or map information) - an ability clearly shown in some migratory birds which respond to virtual magnetic displacements by correcting their orientation (experiments when animals are exposed to magnetic cues corresponding to other geographic locations). Here, we present results from virtual magnetic displacement experiments conducted on red underwings (Catocala nupta). In addition, we tested their orientation under simulated overcast conditions and in a vertical magnetic field to get indications whether this species relies on geomagnetic or celestial cues to maintain its population-specific migratory direction. Our results show that (1) red underwings did not compensate for virtual magnetic displacement, indicating the absence of a magnetic map; (2) they remained significantly oriented in the absence of geomagnetic information, suggesting the use of a stellar compass; and (3) there was no evidence of magnetic compass orientation in absence of any visual cues.

Migration is widely recognized as a strategy for animals to track seasonally shifting resources. Yet, seasonal and spatial dynamics of migration are challenging to study, particularly for difficult-to-track insects. Among insects, monarch butterflies (Danaus plexippus) have a well-documented fall migration, but spring breeding recolonization remains poorly understood, particularly for the western population. We conducted multi-year surveys across six regions in the western United States to characterize monarch breeding phenology and evaluate three related hypotheses: (i) the successive broods model, with discrete generations shifting activity across the breeding range, (ii) a diffusion-like expansion model with overlapping breeding periods, and (iii) a mid-summer lull model with temporary summer declines in breeding for areas near the overwintering habitat. Monarch immature presence served as an indicator of local breeding activity. Our results do not support the successive broods or mid-summer lull hypotheses. Breeding onset occurred earlier near overwintering areas and gradually expanded north-and eastward, with sustained activity in many regions throughout the season. Termination of breeding also occurred earlier at more distant sites, resulting in longer breeding activity nearer to overwintering habitat. Immature monarch density declined with distance from overwintering areas at onset and termination, suggesting delayed colonization of peripheral regions. Together, these results support a diffusion-like expansion of breeding rather than sequential generational replacement. Western monarchs also do not initiate or terminate breeding in close synchrony with host plant availability, contrary to predictions from resource-tracking theory. These findings highlight fundamental differences between western monarch breeding dynamics and paradigms for eastern monarchs, demonstrating that a single species can employ fundamentally different spatial strategies for recolonizing its breeding range in different regions. More generally, these results distinguish insect migration from systems with direct movements between wintering and breeding habitats, and underscore the value of long-term, landscape-scale monitoring for resolving habitat use across heterogeneous environments.

Animals must continually balance foraging with the risk of predation. In complex natural environments, this means quickly distinguishing between threats and harmless situations. We investigated how site-associated coral reef fishes decide to escape in response to visual cues mimicking predator attacks, using controlled underwater presentations of looming stimuli at varying speeds. We measured escape responses across species and social contexts, comparing them to predator attack speeds observed in the same habitat. Escape responses were highly sensitive to the speed of the looming stimulus, with no responses occurring at low speeds. The speeds triggering escape matched those of predator attacks, whereas cruising swim speeds never triggered a response. Species employed distinct antipredator strategies: Brown Chromis foraged away from shelter with high responsiveness, whereas Bicolor Damselfish remained shelter-dependent with lower escape propensities. Contrary to expectations, the social factors did not affect responses in this study. These findings demonstrate that reef fish are highly sensitive to the approach speed of objects, with species-specific strategies further shaping behaviors. By combining realistic visual threats with natural predator attack data, this study offers insight into how animals make escape decisions in complex, real-world environments.

Because Strepsiptera can fly vertically from a standing start, at least [1/4] of their body mass must be dedicated to flight muscle. Adult male Strepsiptera also do not feed and die within a few hours of eclosing, so much normal adult insect anatomy has been discarded, leading to a flight muscle to total mass ratio (FMR) of at least 30%--this is medial for Hymenoptera, but as the lower bound for Strepsiptera, it indicates substantial aerial ability. On account of their high FMR and low wing loading, Strepsiptera are capable of widely varied flight. Moreover, the often incongruous descriptions thereof (that they fly slowly, fly quickly, are clumsy, are graceful, etc.) are paralleled in well-established phases of sex pheromone tracking in moths. For nearly all of their brief eclosed adult lives, male Strepsiptera are airborne, for which they are well-adapted. Correspondingly, strepsipteran propagation is utterly dependent on flight. Thus, flight is the lens through which much strepsipteran ecology is clarified. Accordingly, I photographed free-flying Triozocera texana (nocturnal) in the field and analyzed the images. Strepsipteran wings are remarkably flaccid and potentially teneral, leading to certain flight advantages. At night, spatial acuity is especially poor in tiny insects, but halteres apparently compensate so well that even later derived diurnal Strepsiptera identify calling females chemotactilely--not visually--and shun resolution for high sensitivity. Future directions are discussed, as well as experimental techniques that are problematic when applied to Strepsiptera.

Group foraging can enhance prey detection, but depending on resource availability, it may also generate conflicts among conspecifics. To understand how animals balance these benefits and costs, foraging performance must be evaluated together with inter-individual interactions. However, under fully natural conditions, it remains challenging to quantify both simultaneously. Here, we investigated how individual foraging efficiency and pairwise interactions are shaped when more than one individuals simultaneously exploit the same foraging patch, using the Japanese large-footed bat (Myotis macrodactylus) as a model system. We monitored an entire pond functioning as a natural foraging patch using two thermal cameras and an eight-channel microphone array, and reconstructed the arrival, prey-attack, and exit times of individual bats. Using a Poisson generalized linear mixed model (GLMM), we found that prey-attack rates were approximately 25% lower during paired flights than during solitary flights. We then constructed a null model in which arrival, attack, and departure events followed independent Poisson processes parameterized from the empirical data. Compared with null-model predictions, both the total duration and the duration of individual paired flights in the empirical data were significantly shorter, indicating that bats limited the time spent co-using the same patch relative to solitary foraging. In addition, the probability that the first exiting individual was the one that arrived earlier or later did not deviate from chance levels, providing no evidence for a prior residence advantage. Together, these results demonstrate that simultaneous patch use avoidance occurs independently of arrival order and coincides with reduced prey-attack rate, suggesting that bats leave shared patches and move to alternative foraging sites to mitigate losses in prey-attack efficiency. Our findings highlight bats as an excellent model system for non-invasively linking individual behavior and foraging performance via echolocation, and for elucidating the dynamics of foraging behavior and sensory interference in the wild.

Carausius morosus, the Indian stick insect, is a slender twig-like insect endemic to India. Though widely introduced through captivity around the world and commonly used in laboratories or kept as a household pet, standardized animal husbandry laboratory protocols are lacking. Here we report detailed laboratory culture conditions for C. morosus. We maintain stocks at 23 {degrees}C, 70% relative humidity, and a 12:12 hour light-dark photoperiod. This culture has been successfully sustained under these conditions for over two years, with standardized protocols in place for dietary and cage setup conditions. We also report methods for egg and hatchling care to support ongoing experiments with C. morosus. These standardized methods improve reproducibility and accessibility, enabling the broader use of C. morosus as a laboratory model system for developmental, behavioral, and physiological studies. SummaryThis paper outlines detailed protocols for maintaining a Carausius morosus laboratory colony, including key procedures for animal husbandry, egg and hatchling care, and an overview of the species lifespan and biological characteristics.

O_LIEctotherms in thermally variable environments mediate energy expenditure through both physiological and behavioural responses. However, many studies focus on constant temperature acclimation, and few consider behaviour and physiology in unison. It is unclear how acclimation to thermal variability affects locomotory choices, activity timing, and performance across daily thermal cycles. C_LIO_LIWe investigated the effects of thermal variability in the temperate dung beetle Onthophagus taurus. Following acclimation to a low amplitude (22{degrees}C {+/-} 2{degrees}C) or a high amplitude (22{degrees}C {+/-} 10{degrees}C) temperature regime, we measured behaviour and metabolic rate across temperatures. We hypothesised that O. taurus adjusts its locomotive strategy and search window when kept in high amplitude fluctuating temperatures to reduce energy loss associated with high temperature exposure. C_LIO_LIWe found that differences in energy expenditure were determined by propensity for flight which differed between acclimation treatments, particularly at intermediate temperatures. We also found that, following acclimation to a high amplitude of thermal variability, O. taurus exhibited a greater intensity of activity over a narrower window of time, and O. taurus acclimated to a low amplitude of thermal variability showed nocturnal activity. C_LIO_LIWe then used the data to model activity through the growing season over five years. Biophysical models were built using NicheMapR Microclimate and Ectotherm functions to test the length of potential searching time across seasons, the temperatures individuals are exposed, and locomotive strategy. Model outputs showed that acclimation to higher amplitudes of thermal variability increased accumulated degree-hours of activity relative to the low variability acclimation group. Individuals acclimated to higher amplitudes of thermal variability showed greater accumulated degree-hours in spring and fall, but exhibited shorter periods of activity during summer, with the model predicting increased opportunities for flight. Comparatively, O. taurus from the low variability acclimation treatment showed increased night activity in summer but did not fly. C_LI

The relationship between morphology and ecology is mediated by behavior. We explore this relationship by assessing the link between trophic ecology and the use of prey-specific feeding behaviors in a cichlid fish system. Cichlid diversification features repeated transitions between free-moving prey and attached benthic prey, requiring predators to evolve prey-specific approaches to feeding. Using 2000 Hz video, we characterized feeding behavior on an experimental attached benthic prey in seven species of Mesoamerican heroine cichlid spanning three independent transitions to specialized piscivory and two to specialized benthic-feeding ecology. We investigated the effect of feeding ecology on the behavior and kinematics of benthic grazing, a derived, specialized mode of cichlid feeding. Surprisingly, all species readily fed on benthic prey, regardless of their feeding ecology. Nearly all non-benthic species used the same benthic-feeding behaviors as ecological benthic-feeders. Our findings demonstrate an unexpected level of behavioral versatility among cichlid species in exploiting functionally demanding prey outside their typical diets. We propose that this repertoire of latent feeding behaviors supports trophic versatility and facilitates niche diversification. We also show that two benthic-feeding lineages of Neotropical cichlids evolved distinct approaches to benthic feeding, exhibiting the highest and lowest total feeding-strike kinesis, respectively. Together, our findings highlight the importance of behavior in linking morphology and ecology and motivate further study into the diversity and evolutionary context of benthic feeding across the Cichlidae. SUMMARY STATEMENTWe demonstrate that prey-specific feeding behaviors and strike kinematics vary with trophic ecology in heroine cichlids and discuss the potential role of latent feeding behaviors in trophic diversification.

How insects transmit food odour preferences acquired during the larval stage to their offspring is unknown. Bicyclus anynana butterfly larvae can learn to prefer a banana-smelling odour, isoamyl acetate (IAA), via feeding on coated leaves, or simply via haemolymph transfusions from an IAA-fed animal, and transmit this preference to their naive offspring. Here we explore how larvae respond to different concentrations of IAA using olfaction choice tests, and how injections of different concentrations of IAA directly into the haemolymph impact odour learning and transmission of learned preferences. We find that naive larvae showed a slight preference towards low concentrations of IAA, and a slight avoidance towards higher concentrations. Injections of IAA at low concentrations directly into the haemolymph led to an increase in preference for IAA, whereas higher concentrations led to an increase in avoidance. Naive offspring inherited the odour preferences of their parents. Finally, injections of IAA at different concentrations into embryos did not alter choices made by hatched larvae. We establish that the same molecule (IAA) can illicit both a preference as well as an aversive reaction when directly injected into the haemolymph, but IAA is not directly implicated in intergenerational inheritance.

Historically, venom potencies have been assessed using measures of lethality, such as the median lethal dose (LD50). However, venoms may be selected primarily for their ability to rapidly incapacitate rather than cause mortality, meaning LD50 may not capture the efficacy of venoms in an ecological and evolutionary context. To capture this context, recent studies have adapted measures that assess venoms ability to rapidly incapacitate, such as the median effective dose (ED50). However, while ED50 values are expected to provide a more proximate assessment of ecological variation in venom potency, it is unknown whether historically available LD50 values are still useful proxies of ecologically relevant potency or whether they capture independent axes of venom variation. Here, we test the relationship between LD50 and ED50 in spider venoms by experimentally estimating LD50 and ED50 for 12 species and collating additional potency data for 40 species retrieved from the literature. We observed an isometric relationship between LD50 and ED50 in both analyses, showing these potency measures are both strongly coupled, with an increase in paralysis efficiency associated with a similar increase in lethality. Our results suggest that the functional aspects of venom potency, paralysis and lethality, are intrinsically linked, and due to this strong mechanistic coupling, historically available LD50 values may be used to compare general venom potencies in spiders, provided that they are based on the same prey model.

Edhazardia aedis is an obligate microsporidian parasite of the arthropod vector Aedes aegypti, which is responsible for the spread of several vertebrate pathogens of global health importance. E. aedis can be highly virulent to Ae. aegypti and infection has severely detrimental effects on multiple life history traits that are relevant to the vectoral capacity of Ae. aegypti, including longevity, body size, propensity to host-seek and blood-feed, and reproductive capacity. Because E. aedis is also highly specific to Ae. aegypti and is incapable of completing its full life cycle in any other mosquito species, E. aedis merits investigation as a novel tool for biological vector control. In the present study, we queried the effect of E. aedis infection on the bacterial microbiota of adult female Ae. aegypti using high-throughput amplicon sequencing of the 16S rRNA gene. Analysis of sequencing data revealed that the bacterial microbiota community is strikingly robust to E. aedis infection, as we observed no significant effect on alpha or beta diversity, differential abundance of any taxa, predicted metabolic function profile, or overall bacterial load. The data show that E. aedis, despite dramatically impacting the health and fitness of the adult female mosquito, does not affect the microbiota. These results provide unique insight into tripartite relationships (or lack thereof) between hosts, pathogens, and the microbiota.

Seasonal rainfall shapes biological responses in tropical ecosystems, yet how tropical organisms integrate behavioral and physiological responses to cope with seasonality remains poorly understood. We assessed how four poison frog species with contrasting reproductive strategies respond to dry and wet season environmental conditions. We quantified spatial behavior, microhabitat use, hormone concentrations, and chemical defenses in two seasonal breeders (Allobates femoralis and Ameerega trivittata) and two year-round breeders (Ameerega macero and Ameerega shihuemoy). Seasonal breeders exhibited pronounced sex-specific shifts in space use, where males expanded their space use during the wet season, likely to track reproductive opportunities, while A. femoralis females increased their spatial use during the dry season, likely responding to foraging demands when prey resources are sparse. Year-round breeders maintained similar space use across seasons, likely reflecting their ability to access key resources within the same space to reproduce year-round. Microhabitat use was flexible, as seasonal breeders shifted toward humid refugia during the dry season and reproduction-associated microhabitats during the wet season, whereas year-round breeders selected microhabitats that facilitate continuous reproduction across seasons. Despite these behavioral responses, corticosterone, testosterone, and chemical defenses showed no consistent seasonal variation, suggesting that behavioral flexibility is decoupled from seasonal variation in these measured physiological responses. Our study suggests that poison frogs are able to buffer environmental fluctuations through behavioral flexibility. However, given the increasing unpredictability in rainfall timing and intensity as a result of climate change, how these coping strategies will function in the long term is uncertain.

Extreme climatic events impose considerable stress on organisms with consequences for key ecological interactions such as pollination. Because temperature directly affects metabolic processes, heat variations may also importantly influence the nutritional needs and feeding choices of animals. Here, we studied the effects of thermal stress on the nutritional choices and performances of bumblebees, using a 3D nutritional geometry design. At optimal temperature for colony development (30{degrees}C), bees successfully balanced carbohydrate, protein, and lipid collection, at levels beneficial for body weight and survival. Under cold stress (20{degrees}C), bees reduced their overall nutrient collection while selecting proportionally more carbohydrates, thereby prioritizing survival over weight gain. Under heat stress (35{degrees}C), nutrient balancing was disrupted and survival dropped. Notably however, across all temperatures, bees maintained stable lipid collection while flexibly adjusting the amount of carbohydrates and proteins, suggesting strong constraints on lipid regulation. Given the pivotal role of bees for pollination, identifying how their nutritional needs change in response to climatic conditions is of prior importance for food safety and the conservation of terrestrial ecosystems.

Despite its ubiquity in natural flows, the effects of turbulence on fish locomotion and behavior remain poorly understood. The prevailing hypothesis is that these effects depend on the spatial and temporal scales of the turbulence relative to the fishs size and swimming speed. But in conventional facilities, turbulence usually increases with mean flow, which forces higher swimming speeds and can leave these relative scales unchanged. We therefore present a novel experimental facility that leverages a jet array to decouple the turbulence from the mean flow and systematically control its scales. This approach allows the ratio of turbulent to fish inertial scales to be varied over an order of magnitude, providing a controlled framework for quantifying fish-turbulence interactions. The facility also supports experiments probing strategies fish may use to cope with turbulence, including collective behaviors. Insights from this work have broader implications for ecological studies and engineering applications, including the design of effective fishways and bio-inspired underwater vehicles.

Floral displays attract pollinators through a finely tuned interplay of colour, pattern, shape, and scent. Yet, the question remains: how do bees respond when these traits are stripped to their simplest form, with only visual cues at play? In this field study, we examined the foraging behaviour of Apis mellifera on artificial flowers differing solely in background colour (white or yellow) and UV patterning, while shape and scent were held constant. Across three summer days, standardized stimuli were placed within a natural meadow, and bee-flower interactions were recorded and analyzed by Bayesian hierarchical models. The results reveal a clear preference for yellow over white backgrounds and prolonged visitation in the presence of ring-shaped UV patterns, whereas full UV coverage acted as a deterrent. These effects, though moderate, were consistently modulated by abiotic covariates, particularly radiation, temperature, and time of day. Negligible inter-individual variation and a substantial share of residual variance further underline the context-dependent complexity of foraging. In sum, our findings demonstrate that visual floral traits, while influential, are interpreted through the dual lens of environmental contingency and the bees inherent cognitive machinery.

Environmental change is reshaping wildlife reproduction through increasing temperatures and the spread of emerging infectious diseases, yet the physiological consequences of managing these stressors remain poorly understood. Amphibians are particularly vulnerable due to their ectothermy and high susceptibility to chytridiomycosis, caused by Batrachochytrium dendrobatidis (Bd). Here, we examine how Bd infection and thermal treatment interact to influence sperm quality and reproductive investment in male green and golden bell frogs (Ranoidea aurea), a species that has suffered severe population declines. Moderate Bd infection was associated with elevated sperm concentration relative to uninfected and heavily infected males, consistent with increased short-term reproductive investment under elevated mortality risk. However, severe infection led to pronounced reductions in sperm concentration and motility. Thermal treatment successfully eliminated Bd infection but imposed substantial reproductive costs: sperm concentration declined following treatment and remained significantly reduced six months later, despite partial recovery of sperm motility and membrane integrity. These results indicate persistent impairment of spermatogenic capacity rather than transient suppression. Our findings reveal that disease and thermal stress jointly shape amphibian reproductive outcomes through context-dependent trade-offs between immune defence and gamete production. While mild infection may trigger short-lived increases in reproductive output, both severe infection and pathogen clearance via thermal exposure impose lasting constraints on fertility. These results highlight an underappreciated cost of disease mitigation and suggest that increasing thermal extremes associated with climate change may further limit amphibian reproductive resilience, with important implications for conservation management and population persistence.

Understanding how fish navigate complex natural environments requires bridging fine-scale biomechanics with ecological behavior. We investigated the volitional movement and energetics of wild red drum (Sciaenops ocellatus) across laboratory, mesocosm, and field settings. Using flow-respirometry, we quantified metabolic costs and swimming kinematics under ecologically relevant flow conditions shaped by bluff bodies mimicking mangrove roots and oyster mounds. Fish swimming in turbulent wakes exhibited reduced oxygen consumption and altered tailbeat dynamics, especially at high flow speeds. In a large outdoor mesocosm, dual accelerometers revealed a rich behavioral repertoire, including maneuvering and rest, which is not easily observable in confined lab settings. Spectral analysis and clustering identified eight distinct locomotory states, highlighting the limitations of summed acceleration metrics. Field telemetry tracked wild red drum across a 54 km estuarine corridor for a three-year period through an array of 36 acoustic receivers, revealing movement patterns shaped by tidal flow and physical habitats. Hydrodynamic modeling revealed that while laboratory trials demonstrated substantial energetic savings at high flows (approaching 100 cm/s), wild fish were detected predominantly in low-velocity microhabitats (<30 cm/s) near structurally complex features. This mismatch suggests that habitat selection is an adaptive strategy driven by ecological factors such as foraging opportunities, predation refuge, and site fidelity, rather than hydrodynamic efficiency alone. Our multi-scalar approach demonstrates that while flow-structure interactions can reduce locomotor costs for fish, habitat use in the wild reflects broader ecological constraints, offering a framework for integrating biomechanics, physiology, and ecology in conservation-relevant contexts.