Integrative And Comparative Biology

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.

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.

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.

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.

Iron is an essential nutrient for all types of organisms, including insects and the microbes that infect them. We predicted that insects fed an iron-supplemented diet would accumulate more iron in their hemolymph, and, because infectious microbes acquire iron from their hosts, that this extra iron would increase the severity of bacterial infections. To test this hypothesis, we studied the effects of dietary iron supplementation on infection outcomes in Manduca sexta (tobacco hornworm). Larvae were fed an artificial diet, with or without antibiotics, or the same diets supplemented with 10 mM iron. Control and iron-treated larvae were inoculated with non-pathogenic Escherichia coli or the entomopathogenic Enterococcus faecalis, and bacterial load and larval survival were measured. We found that dietary iron supplementation increased the iron content of hemolymph by approximately 20 fold; however, contrary to our prediction, this increase in iron did not result in an increase in the bacterial load of either E. coli or E. faecalis. The effect of iron supplementation on survival was more complicated. As expected, for larvae inoculated with nonpathogenic E. coli, iron supplementation had no effect. For larvae inoculated with E. faecalis, the effect of iron supplementation depended on whether antibiotics were present in the diet. Without antibiotics, iron supplementation prolonged larval survival; with antibiotics, iron supplementation decreased larval survival. The results of this study do not support the hypothesis that dietary iron supplementation increases infection severity in M. sexta. Instead, the results support the viewpoint that the relationship between dietary iron and infection outcome is complex.

Although predation is a major driver of group living across taxa and the antipredator benefits of grouping are well established, the energetic costs experienced by groups under predation remain largely unexplored. In the current study, we use wild, white mullet (Mugil curema, Valenciennes 1836), to provide the first real-time quantification of the energetic cost of escape in schooling fish using intermittent, closed-loop respirometry. We found that small groups exposed to predators showed a 53.8% increase in their organismal metabolic rate (MO2) as compared to groups without predator exposure. When we evaluated antipredator behaviors such as escape response, group cohesion, and displacement of the group centroid, we found a positive correlation to energetic costs. We then investigated whether escape responses are socially modulated by comparing the energetic costs of escape across solitary individuals, solitary individuals with visual access to a group, and groups. We found that escape frequency and energetic costs to predation were comparable across social contexts, indicating that escape may be an intrinsic survival response independent of cues from group members. Furthermore, we found that fish exposed to predators showed markedly reduced feeding, suggesting that predation constrains energy acquisition in addition to imposing direct energetic costs. Our results provide the first direct quantification of the energetic costs of escape in a schooling fish, offering new insights into the physiological trade-offs underlying collective antipredator defenses.

Sea star wasting disease has caused widespread mortality in the kelp forest predator, the sunflower sea star (Pycnopodia helianthoides). Wild populations have declined by up to 99% in parts of their native range along the western North American coast. In response, a multi-institutional conservation breeding and rearing program has been initiated to support future reintroduction efforts for the species. We split a full-sibling cohort across four larval density treatments (1 larva/ml, 2 larvae/ml, 5 larvae/ml, and 15-20 larvae/ml) to assess the effects on larval settlement, juvenile survival, and juvenile fitness at 12 months old. Stars raised in the highest density treatment displayed a lower settlement rate and were significantly smaller than the other density groups at 12 months old, but showed no significant difference in flip time, a measure of fitness. Additionally, measurements of diameter, weight, and arm count across modern and historical juvenile and adult stars indicate that P. helianthoides experience exponential weight gain as they grow in length, with corresponding asymptotic growth in arm count. These findings will inform best practices for the aquarium propagation of P. helianthoides and will contribute to broader efforts aimed at reestablishing populations in the wild.

Collective behavior in animal societies can buffer individual costs and confer resilience to environmental challenges. However, the mechanisms by which groups sustain function when members are compromised remain poorly understood. In the presented study, we investigate how social context shapes collective fanning, a thermoregulatory behavior critical for colony function, in Western honeybees (Apis mellifera). Using oxytetracycline (OTC), a known physiologically disruptive antibiotic to honeybees, to selectively impair certain group members, we tested our hypothesis that the presence of untreated bees would rescue the fanning response in mixed-composition groups. We show that groups containing untreated individuals fan at levels comparable to fully untreated groups, despite the presence of OTC-impaired bees. This preservation of collective thermoregulatory function was correlated with both treated and untreated individuals in mixed groups shifting their interaction dynamics and social network positions. These findings reveal a decentralized mechanism of collective resilience, whereby behavioral compensation by individuals sustains group-level thermoregulation under partial disruption. Our results provide a framework for understanding how social insect colonies maintain function in the face of individual-level perturbations, with broader implications for predicting the limits of collective resilience in animal societies experiencing increasing environmental pressures.

Protogynous sex change, where individuals first function as females and later as males, is a key life-history strategy among polygynous reef fishes. In haremic systems, sex change is typically socially regulated, with dominants suppressing subordinates sex change through aggression. Females within a harem form a size-based hierarchy that can remain stable in most species through the threat of eviction. We studied a different situation in the cleaner wrasse Labroides dimidiatus, where larger females have incomplete control, as they spend most of their time alone at their own cleaning territory. We tracked over 400 individuals for 12 months, recording growth, behavior, social organization, and sex change. We confirmed earlier reports that both sexes direct aggression primarily at those ranked immediately below them. However, we observed 30 cases where smaller females outgrew larger ones, revealing hierarchy instability. Of 42 sex change events, 43% occurred in presence of the male, and half of these early sex changers were not the largest female, but individuals overlooked by the male. Fast growth relative to harem-mates and harem switching increased the likelihood of sex change. Local population densities also influenced growth and sex change, with individuals in high-density demes growing faster and changing sex at larger sizes. Our findings reveal flexible sex change dynamics in a system with incomplete social dominance. Such incomplete control and observations that becoming male confers both higher reproductive success and survival highlight the need to expand game-theoretical and life-history frameworks to encompass such strategic flexibility. Lay summaryDominant cleaner wrasse cannot fully control subordinates as individuals occupy distinct core areas. Tracking 400 fish for a year, we found that smaller females could outgrow initially larger ones, and early sex change despite a larger male. Fast growth and harem switching increased the chances of becoming male. Population density also shaped these strategies. Our findings reveal flexible sex change dynamics in a system where becoming male confers both higher reproductive success and survival.

Sentinel behaviour occurs when individuals use raised positions to scan for predators while the rest of the group forages. Here, we investigated whether a colonial cooperatively breeding species that forages in large groups, the sociable weaver, Philetairus socius, displays sentinel behaviour. This behaviour has been reported in species with similar ecology, behaviour and foraging habits, (e.g. ground foraging in open habitats where aerial predators are common) and, hence, we expected that it could occur in sociable weavers. On the other hand, sentinel behaviour appears to be less common in species that live in very large groups. We used an experimental set-up consisting of an artificial feeding station and perches to assess occurrence of sentinel related behaviours: (i) perching events > 30s on an elevated position, (ii) head-movements and (iii) alarm calling. Birds were seldom observed perching while others fed, and those that did, perched for periods that were too short to be considered as sentinel behaviour (less than 5s on average). Our results suggest that this behaviour is uncommon or even absent in sociable weavers. We discuss whether other factors such as foraging in very large groups, or interspecific foraging associations might make sentinel behaviour less important in this species.

O_LIPhenological shifts are a major ecological consequence of climate change, yet studies often focus on single life stages meaning that the potential for carryover effects between life stages remains poorly understood. Failing to account for these effects may lead to inaccurate estimates of phenological shifts, with consequences for predicted synchrony among interacting species. This is especially relevant for temperate systems where climate warming is occurring unevenly across the year. C_LIO_LIHere, we investigated how temperature experienced the previous autumn and winter (during the pupal and egg stage) influences spring phenology in the winter moth (Operophtera brumata), a herbivorous insect with distinct life stages. Using 50 years of local climate data to create five experimental temperature regimes, we first quantified phenotypic plasticity in the duration and temporal variability of pupal and egg development. We then examined how timing of adult moth emergence affects timing of offspring hatching. C_LIO_LIWe found divergent effects of temperature on different life stages; pupal development time was shortest at intermediate temperatures while egg development time decreased linearly with increasing temperature. Furthermore, phenological shifts due to the conditions experienced by the mother were carried over to influence the phenology of her offspring. While this carryover effect was partially compensated during subsequent stages, compensation decreased under warming conditions. C_LIO_LIThese results refine our understanding of the sensitivity of the annual cycle of winter moth phenology to variation in temperature with potential implications for population dynamics and interspecific interactions. Overall, our findings highlight the need to consider the impacts of warming across multiple life stages so that carryover effects can be properly accounted for. Doing so will improve predictions of phenological shifts under future climates. C_LI

Animals sense and integrate complex external cues to make developmental decisions that help them better survive and adapt to their natural habitats. Under environmental adversity, nematodes can enter an alternative developmental pathway to form a diapautic and stress-resistant stage, termed the dauer larvae. While dauer formation has been well characterized in Caenorhabditis elegans, how environmental factors influence analogous stages in other nematode species remains largely unexplored. This study examines how symbiotic bacteria, temperature, and pheromones affect the formation of the infective juvenile (IJ), a dauer-like stage, of the insect-parasitic nematode Steinernema hermaphroditum. In contrast to C. elegans, where dauer entry is promoted by heat, IJ development in S. hermaphroditum development is enhanced by reduced temperature. Moreover, the presence and absence of live symbiotic bacterium Xenorhabdus griffiniae functions as an ON-and-OFF switch that regulates the host IJ formation. Crude pheromone extracts from S. hermaphroditum liquid culture do not robustly induce IJ formation in a dose-responsive manner, unlike the potent pheromone-driven dauer entry observed in C. elegans. Nutrient-rich liver-kidney media that mimics host insect environment showed IJ entry induction in a pheromone-dependent manner. These data suggest that external cues, such as temperature, microbial diet, and pheromone, are perceived differently by S. hermaphroditum in comparison to that of C. elegans, reflecting species-specific adaptations to distinct ecological niches and life history strategies.

Understanding habitat association of animals and how they change through ontogeny is critical to predict the likely effects of habitat change on populations. We investigated how fine scale habitat associations of three common coral reef damselfish species changed among life-stages on reefs surrounding Lizard Island, northern Great Barrier Reef. All three species showed distinct habitat selection at settlement, however the degree to which these initial associations changed through ontogeny were species specific. Pomacentrus amboinensis associated with sandy areas throughout all life-stages; Pomacentrus chrysurus settled to areas with high cover of sand and rubble, but displayed no clear habitat preferences as juveniles or adults. Pomacentrus moluccensis settled to areas with high cover of fine branching corals before shifting to areas with relatively high cover of soft corals as adults. We also compared two different approaches to estimate habitat selection; one that quantified the benthic composition within the approximate home range of individuals versus a more widely used approach of recording a single point underneath the focal individual when they were first observed. Although results were broadly similar, the benthic composition approach revealed details that was overlooked using the single point method. Decreases in the availability of any of these preferred benthic habitats may adversely affect future populations, therefore understanding habitat associations and their transitions among life stages will be crucial in predicting future reef fish communities under ongoing coral loss and habitat change. This will require to systematically study a broader range of species, integrating relevant spatial and temporal scales.

Stingless bees are key pollinators in tropical ecosystems, yet their ecological dynamics remain poorly understood in highly seasonal environments such as the seasonally dry tropical forests of Ecuador. These ecosystems experience pronounced climatic seasonality, with sharp transitions between dry and wet periods that strongly affect floral resource availability. Understanding interspecific competition and niche partitioning in such systems is critical, particularly given the global decline of pollinators. We investigated resource use and niche dynamics in two native stingless bees, Melipona mimetica and Scaptotrigona sp., by quantifying pollen, nectar, and resin collection across seasons. Log-linear models were used to test the effects of species, season, and their interaction on resource use, while non-metric multidimensional scaling (NMDS) assessed niche overlap. Contrary to the expectation that niche overlap increases under resource scarcity, we found greater overlap during the wet season, when resources are more abundant. This suggests that both species converge on high-quality floral resources during peak availability, reflecting an adaptive response to strong environmental seasonality. Pollen use remained stable across seasons, consistent with generalist foraging behavior. In contrast, nectar collection increased significantly during the wet season, while resin exhibited a shared seasonal peak, likely associated with synchronized nest construction or maintenance. These findings reveal context-dependent competition dynamics and highlight the role of environmental seasonality in shaping pollinator interactions. Our study provides new insights into the ecology of threatened stingless bees and contributes to their conservation in tropical dry forest ecosystems.

The coleoid cephalopods (octopus, cuttlefish, and squid) are emerging model organisms for neuroscience, development, and evolutionary biology. Determining their sex early in life is critical for population management and controlled experiments. Here, we present a protocol to non-invasively determine the sex of multiple cephalopod species as young as 3 hours post-hatching using a skin swab and quantitative PCR (qPCR). We describe steps for designing qPCR primers, swabbing live animals, extracting DNA, running the qPCR, and analyzing the results. For complete details on the use and execution of this protocol, please refer to Rubino et al.1 HighlightsO_LISwab live cephalopods as early as 3 hours post-hatching C_LIO_LIExtract DNA from cephalopod skin swabs C_LIO_LIPerform qPCR-based sex determination C_LIO_LIDesign and validate qPCR primers for new species C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=190 SRC="FIGDIR/small/715692v1_ufig1.gif" ALT="Figure 1"> View larger version (43K): org.highwire.dtl.DTLVardef@3aa68dorg.highwire.dtl.DTLVardef@8c7e61org.highwire.dtl.DTLVardef@1bd45d9org.highwire.dtl.DTLVardef@134cc4d_HPS_FORMAT_FIGEXP M_FIG C_FIG

In many polygynous species, males face stronger intrasexual competition, higher energetic demands, and lower survival than females, especially under resource limitation or environmental stress. Such sex-specific vulnerabilities are expected to intensify with climate change. Yet, in sequentially hermaphroditic systems, where individuals change sex during their lifetime, how sex and sex change shape survival remains largely unexplored. We studied sex-specific survival and growth in the haremic protogynous cleaner wrasse Labroides dimidiatus across eight reefs around Lizard Island, Great Barrier Reef. We tracked a total of 731 adult fish (individually recognizable through marking or idiosyncratic color patterns) over two years. This period included the 2024 El Nino-Southern Oscillation (ENSO), which caused a temporary 1-degree increase in water temperature, severe coral bleaching, and coral mortality at Lizard Island. Contrary to expectations from dioecious systems, terminal-phase males exhibited higher survival than initial-phase females under both normal and in particular ENSO conditions. While male mortality was not affected, female mortality more than doubled during the event, indicating greater physiological or energetic vulnerability. A partial explanation for the overall higher female mortality is their generally faster growth rate, which declined in both sexes during the ENSO event. Our findings challenge existing assumptions of male-biased mortality in polygynous species and highlight that sex and sex change fundamentally shape demographic responses to climate extremes.

Population density can influence individual predation risk in colonial breeders through shared vigilance and predator deterrence. We investigated how predator-prey interactions are shaped by population density at two Antarctic fur seal (Arctocephalus gazella) breeding colonies at Bird Island, South Georgia, which differ four-fold in seal density. By deploying autonomous time-lapse cameras, we captured high-resolution images at one-minute intervals throughout the breeding season. Using a YOLOv8 neural network, we identified fur seal adult males, females and pups, as well as three predator-scavenger bird species: giant petrels (Macronectes spp.), brown skuas (Stercorarius antarcticus) and snowy sheathbills (Chionis alba). Abundance patterns corresponded to the known foraging and breeding behaviours of these species. Differences in seal density between the colonies were mainly driven by adult females and their pups, but not adult males. The ratios of predatory birds to pups were markedly lower at the high-density colony, while scavenger to pup ratios remained similar. Spatial analyses revealed that predators were largely excluded from areas of high seal density, whereas scavengers overlapped extensively with pups in both colonies. This study demonstrates the value of remote observation in resolving predator-prey interactions and illustrates how density can shape predation risk in a colonial breeder.

Predator-prey interactions are key drivers of behavioural and life-history evolution, yet their mechanisms remain difficult to study in natural contexts. The nematode Pristionchus pacificus is a model predator, but most studies exploring its behaviours use Caenorhabditis elegans as prey, a species that it likely only rarely encountered in nature. Here, we examine predation within nematode communities associated with beetle carcasses, the native necromenic habitat of P. pacificus. We identify Oscheius myriophilus as a cohabiting species, likely representing natural prey. Using predatory assays, automated tracking, and machine-learning-based behavioural analysis, we show that P. pacificus actively kills and consumes O. myriophilus. Strikingly, predation rates are lower than those observed for C. elegans, suggesting partial resistance or reciprocal adaptation in O. myriophilus. Consistent with this, O. myriophilus exhibits a mixed reproductive strategy, with early oviposition followed by ovoviviparity and matricide. As later developmental stages are more resistant to predation, internal hatching may protect offspring while providing maternal resources for development. These findings establish these nematodes as a tractable model for investigating predator-prey interactions and their evolutionary consequences, highlighting how behavioural strategies and life-history traits can co-evolve in natural communities.

Large commercial and military aircraft can operate in a wide range of turbulent conditions, except during extreme weather events such as cyclones. Smaller man-made vehicles, such as micro aerial vehicles (MAVs) and nano aerial vehicles (NAVs), are significantly more sensitive to routine environmental wind fluctuations, making them difficult to control. In contrast, insects exhibit remarkable stability in naturally gusty conditions. Despite this, few studies have systematically investigated the impact of gusts and turbulence on insect flight performance. To address this gap and to gain fundamental insights into insect flight stability under gusty conditions, we examined the flight of freely flying black soldier flies subjected to a discrete head-on aerodynamic gust in a controlled laboratory environment. Flight motions were recorded using two high-speed cameras, and body and wing kinematics were analyzed across 14 distinct cases. In response to the gust, we observed consistent features across the cases: (1) asymmetry in wing stroke amplitude, (2) large changes in body roll angle--up to 160{degrees}--occurring over approximately two wing beats ([~]20 ms) with recovery over [~]9 wing beats, (3) transient pitch-down attitude, and (4) deceleration in the flight direction. These rapid responses, combining passive and active control mechanisms, provide insight into the flight control strategies employed by insects. The findings offer valuable guidance for the design of MAVs and NAVs capable of robustly responding to gusts and unsteady airflow in natural environments.