Integrative and Comparative Biology

Extensible body structures have evolved repeatedly across animals, yet the mechanisms underlying the deployment of extensible organs often remain unknown. Eristalinae hoverfly larvae (rat tailed maggots) possess exceptionally elongated posterior respiratory siphons, but the mechanism underlying their extension has not been experimentally investigated. Here, using wild collected Helophilus virgatus larvae, we show that posterior siphon extension is achieved through a folding unfolding mechanism revealed by fluorescence labeling. Phalloidin staining further demonstrated that, unlike Episyrphus sp. and Drosophila melanogaster, H. virgatus possesses a dense array of transversely oriented muscle fibers in the posterior siphon. Behavioral analyses further revealed that the posterior siphon functions not only in respiration but also as a propulsive organ for near surface locomotion through asymmetric rowing. Together, our findings identify the structural and kinematic basis of posterior siphon deployment and demonstrate how a specialized respiratory organ can evolve into a multifunctional appendage that supports both respiration and locomotion.

Predator-prey interactions shape the evolution of escape behavior in prey, including different combinations of evasive movements, that may enhance unpredictability in fleeing directions and trajectories. So-called protean motion can enhance survival of flying prey in the wild, but quantifying such behaviors under natural conditions remains challenging. Here we used stereoscopic high-speed videography to record the escape flight behavior of wild males of the butterfly species Morpho menelaus in the Amazonian rainforest, and reconstructed 3D flight trajectories using artificial-neural-network-based tracking. During the experiments, we used a lure to attract freely patrolling male butterflies and elicited escape flights by intercepting their trajectory with a looming insect net swing. We then compared the escape flight kinematics to the pre-attack patrolling behavior. Attacks first induced a rapid upward maneuvering, directly followed by an unpredictable horizontal turn. The following escape flight trajectories showed increased horizontal erraticity and greater intra-individual heading variability, as compared to the pre-attack flight. Surprisingly, the mean speed decreased in the escape phase, notably in the horizontal plane. A significant negative association between horizontal trajectory complexity and flight speed was detected, indicating a speed-erraticity trade-off. These results show that wild Morpho butterflies respond to attacks by combining a climbing maneuver with an unpredictable heading change, followed by a protean escape flight; this increased escape erraticity comes at the expense of reduced escape flight speed. Because these large and relatively slow-flying butterflies display bright iridescent blue coloration on their dorsal wing side, erraticity during flight might enhance the dynamic flash coloration, likely limiting accurate targeting by predators.

Robotic herding offers a promising off-animal approach to controlling the spatial distribution of grazing livestock. Nevertheless, the habituation of cattle to herding stimuli remains a critical obstacle to its practical use. This study presents the first field investigation of behavioral responses of grazing cattle to herding by a quadruped robot. We compared the herding efficacy of a drone and a quadruped robot, and tracked daily changes in responsiveness during continuous herding. Trials were conducted in a 1.1-ha pasture with Japanese Black breeding cows from June to October 2023. The quadruped robot elicited stronger avoidance responses with shorter latencies than the drone, and herding was more successful with the robot. During consecutive daily herding with the robot, behavioral responsiveness declined progressively over the initial 5 days, at a markedly slower rate than previously reported for drone herding. Following a 24-day interruption, responsiveness partially increased, but declined rapidly again over the subsequent two days. These results indicate that the quadruped robot constitutes a more persistent aversive stimulus than a drone for grazing cattle, although habituation management strategies (such as diversifying stimuli or combining sensory modalities) will be necessary for sustained control of grazing distribution.

Many arthropods (insects and crustaceans) rely on their antennular chemosensory system to detect key environmental resources like food. While odor mediated food search is well studied in insects, characterization of crustacean chemosensory behavior has been limited by the long lifespans and large size of traditional crustacean model species. Here, we report the first characterizations of the food search behaviors of the genetically tractable amphipod crustacean, Parhyale hawaiensis. We find that Parhyale can locate an odorous food pellet, and predominantly approach food using direct, targeted swims from the arena walls. Removal of both first and second antennae dramatically reduced foraging success and impaired Parhyales ability to control take-off angle and maintain a stable heading during swims. Removal of the first or second antenna alone did not significantly disrupt foraging, and resulted in mild disruption of orientation phenotypes. Intact animals performed sharp turns near the location of the food pellet, which were observed when either first or second antenna were present, but not when all antennae were removed. Turns were longer and had higher average angular velocities following removal of either set of antennae, with full antenna removals representing the most extreme phenotype. In contrast with the long-held theory that the crustacean second antennae exclusively mediate contact chemosensation, we report that first- and second- antennae both contribute similarly to food localization and stabilization of locomotion in Parhyale in our behavioral paradigm. This work establishes Parhyale as an accessible model for studying olfactory behaviors in an aquatic arthropod.

Carnivorous Nepenthes plants capture and digest prey in fluid-filled pitchers, whose fluid level is subject to fluctuations caused by rain and evaporation. It was found that N. rafflesiana pitchers can reduce these fluctuations by regulating their fluid volume and composition. Here, we investigate whether and to what extent this ability occurs in other Nepenthes species varying in morphology, trapping mechanisms, habitat, lifespan and nutrient sources. We hypothesised that species more exposed to fluid level fluctuations or relying more on fluid for prey capture would exhibit more intensive regulation. In six Bornean Nepenthes species, we quantified the effect of pitcher fluid level on prey-capture efficiency, the natural fluctuations in fluid volume, and the plants response to manipulations of the fluids volume, concentration and pH. The prey capture of Nepenthes species depended to varying degrees on the pitcher fluid level: in species with waxy inner walls, capture efficiency decreased at high fluid levels, as prey could escape easily. In contrast, species with wax-free walls and steep peristomes retained prey efficiently, regardless of fluid level. Although all Nepenthes species were able to secrete and absorb water and electrolytes, the regulation rate varied considerably. The rates of the various secretion and absorption processes (concentration/volume-dependent rates of electrolyte/water transport) were positively correlated with one another, suggesting that they all depend on the activity of the pitcher glandular epithelium. While differences and similarities in fluid regulation may be partly due to phylogeny, the observed species differences can also be explained by the pitchers trapping mechanisms and ecological factors. We found weaker regulation in Nepenthes species growing in sheltered understorey habitats, and in species utilising alternative nutrient sources, which may reduce their reliance on pitcher fluid. Our study highlights the diversity of ecological strategies in pitcher plants and demonstrates how these are influenced by environmental conditions. Lay SummaryNepenthes pitcher plants have fluid-filled pitchers to capture and digest prey. Can they control the volume and composition of their fluids? We found that all six species studied regulated their fluids, but to varying degrees. Species differences can be explained by the plants habitat, pitcher morphology and nutrient acquisition strategies.

Coral reefs are declining globally, creating an urgent need for scalable technologies that improve the efficiency and effectiveness of active reef restoration. Coral reef restoration is increasingly constrained by algal overgrowth, which suppresses coral growth, survival, and restoration efficiency. Building on the recently developed Coral Guard platform, we evaluate its restoration performance under long-term in situ coral nursery conditions and introduce Fusion Guard Tiles, a geometry-optimized Coral Guard design that accelerates microfragment fusion. We evaluated Coral Guard Plugs using the branching coral Stylophora pistillata under complementary ex situ conditions and Fusion Guard Tiles using the massive reef-building coral Porites evermanni in in situ coral nurseries. In P. evermanni, Fusion Guard Tiles increased lateral tissue growth 2.6-fold and three-dimensional tissue surface area growth by >2.7-fold after 6 months compared with conventional substrates. After 12 months, colony height and volume were approximately 4.0-fold and 2.2-fold greater, respectively, while complete fragment fusion occurred only on Fusion Guard Tiles. In S. pistillata, Coral Guard Plugs increased lateral tissue growth by [~]1.7-fold. Microcomputed tomography revealed 10-13% higher skeletal density in both species. In P. evermanni, Fusion Guard Tiles also increased symbiont density by 70% and tissue protein content by [~]2.5-fold relative to controls. Together, these findings demonstrate that Coral Guard substrates suppress algal competition while accelerating coral growth, skeletal development, and microfragment fusion, providing a scalable, low-maintenance technology to enhance coral nursery productivity and reef restoration.

0.O_LIMorphological traits such as floral area and body size are fundamental to ecological research, serving as inputs for studies of pollinator-plant interactions, habitat quality, and biodiversity monitoring. However, accurately measuring these traits from images remains challenging, particularly in complex field conditions where existing tools exhibit reduced accuracy and limited generalizability across taxa. C_LIO_LIWe present EcoMorph, a modular morphological measurement system that leverages the Segment Anything Model 3 (SAM3) to quantify traits across diverse ecological contexts. Unlike task-specific segmentation models requiring domain-specific training data, SAM3s prompt-based architecture enables segmentation of arbitrary biological structures from natural-language prompts, using the same underlying model across flowers, insects, and other targets without retraining. From the resulting segmentations, EcoMorph extracts three classes of measurement: area, linear dimensions, and object counts. C_LIO_LIWe validated EcoMorph across two ecological scales. At the intermediate scale, EcoMorph-derived floral area agreed closely with manual ImageJ measurements (R2 = 0.935, n = 74) under simple-background conditions and (R2 = 0.928, n = 58) under complex-background conditions, with valid predictions for 95% of images. At the fine scale, EcoMorph-derived insect body area was strongly correlated with hand-measured intertegular distance (r = 0.810, n = 349), capturing body-size variation across species from the small Bombus impatiens to the large Xylocopa virginica. Object counts matched manual counts almost exactly for well-separated insects in an insect box (R2 = 0.9997, n = 12). C_LIO_LIBy combining prompt-based segmentation with modular measurement, EcoMorph enables high-throughput quantification of area, size, and abundance from heterogeneous image sources without taxon-specific training. This generality supports a broad range of ecological applications, including pollinator and plant trait research, biodiversity and abundance monitoring, and allometric biomass estimation. C_LI

Animals balance staying in a favorable environment with exploring new ones. In C. elegans chemotaxis, the process by which worms migrate toward an attractant has been extensively studied. However, what happens after they reach it remains largely unexplored, partly because conventional assays immobilize worms at the point of arrival. Here, we quantitatively analyzed chemotactic behavior upon reaching an attractive odor source using an immobilization-free chemotaxis assay. We observed that 62% animals left the isoamyl alcohol region after initially approaching it, a behavior we termed "leaving behavior." Quantitative analysis revealed that leaving behavior represents a distinct locomotor state compared with free-moving, high-concentration odor avoidance, and approach behavior. To test whether leaving behavior is related to olfactory adaptation, we analyzed mutants in adaptation-related genes. The proportion of leaving behavior was significantly increased in egl-4 loss-of-function mutants compared with wild-type animals, whereas arr-1 mutants showed no significant difference. These results suggest that egl-4 negatively regulates leaving behavior, suggesting a role for this kinase in stabilizing post-arrival behavioral states beyond its known function in olfactory adaptation. Our findings indicate that chemotaxis involves dynamic behavioral transitions even after reaching an attractant, consistent with an exploration-exploitation trade-off framework.

O_LISolitary bee species that use artificial trap nests are important for agricultural crop production and as indicators of habitat quality. Quantifying cavity-nesting solitary bee foraging and nesting behavior is essential for real-time analysis of population numbers and pollination activity, as well as understanding how environmental conditions shape reproductive success and population dynamics. However, manual observation is labor-intensive, prone to observer bias, and unable to deliver continuous data. Existing automated systems either require individual bee marking or detect presence without resolving nest-tube-level entry and exit events. C_LIO_LIWe developed BeeMonitor, an integrated hardware and computer-vision pipeline that detects nest entry and exit events in cavity-nesting solitary bees from continuous video, using Osmia cornifrons (the horn-faced mason bee) as a model system. A low-cost Raspberry Pi handles solar-powered field recording, while the software combines object detection (YOLOv26), a custom multiple-object tracker (BeeTrack), and a Random Forest classifier trained on trajectory-derived features to distinguish genuine events from incidental detections. C_LIO_LIOver a 29-day deployment, hardware reliability averaged 97.5% recording coverage. The pipeline achieved 91.3% precision and 87.3% recall (F1 = 0.893), generalizing robustly under leave-one-video-out cross-validation (mean F1 = 0.904). Detected foraging trips correlated strongly with brood cell counts (R2 = 0.849, p < 0.001, n = 19), and a Random Forest model (AUC = 0.820) identified solar radiation as the dominant driver of foraging activity, followed by temperature. C_LIO_LIBeeMonitor demonstrates that automated computer vision can reliably extract ecologically relevant behavioral data from continuous video, enabling real-time analysis of pollinator behavior and abundance at a temporal and spatial resolution unattainable through manual observation. Its modular design supports adaptation to other species and monitoring contexts. C_LI

Argentine ants (Linepithema humile) utilize adhesive pads (arolia) to climb smooth surfaces. Previous research found that ants can adjust their individual arolium engagement according to their locomotion mode. However, it remains unclear how they distribute arolium engagement across multiple limbs to climb effectively, and how arolium engagement varies within a climbing step. As the arolium is a well-known adhesive organ, we hypothesized that engagement across different legs is distributed according to the normal forces required for balancing the body during climbing. To test this, we measured Argentine ants' arolium engagement on a vertical glass surface using a Frustrated Total Internal Reflection (FTIR) sensor and compared it to the required normal forces from a quasi-static model. Contrary to the required normal force, the measured arolium engagement was asymmetric between upward and downward climbing, and changed over time. Our results indicated that the quasi-static force requirements are not sufficient to explain arolium engagement in climbing Argentine ants, and suggested that other factors, such as body dynamics, ants' anatomy and behavioral preferences, should be included.

Urban habitats can serve as wildlife refuges, or as ecological traps. As global urbanization continues to increase, native prey species may experience reduced threat from natural predators that tend to avoid human-populated areas. However, such protection often comes at the expense of susceptibility to synanthropic predators, one of the most prevalent being domestic house cats (Felis catus). While cats are a known driving force of mortalities in urban wildlife, their sublethal effects remain understudied. To better understand the behavioral effects of synanthropic predators on native urban prey, we examined responses of western fence lizards (Sceloporus occidentalis) to Artificial Intelligence (AI)-generated visual stimuli of native predators (snakes) and synanthropic predators (house cats). Visual stimuli included: California striped racers (Masticophis lateralis), a native predator; domestic cats, a synanthropic mammalian predator (Felis catus); and desert cottontails (Sylvilagus audubonii), a native mammalian non-predator. We quantified lizard responses as the change in rates of looking and locomotion during and following the video presentation. Our results revealed that lizards discriminated cats from the control by looking more within the first 90 seconds following exposure. Lizards looked more across the entire 5-minute observation period following exposure to all stimulus types compared to control conditions, indicating that AI generated visual stimuli effectively elicit behavioral responses. Together, these findings indicate that urban lizards initially detect and respond to visual cues of cats, suggesting that cat-induced mortality is not a result of inability to detect such predators. Thus, cats influence urban prey not only through direct predation risk, but also changes in behavior associated with the presence of synanthropic predators.

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.

Collective movement in social organisms emerges from local interactions and can generate large-scale spatial patterns of ecological relevance. In termites, trail formation is a well-known collective phenomenon, yet reproducing and recording its emergence under controlled laboratory conditions using whole colonies remains challenging. Existing laboratory approaches often rely on confined arenas or manually assembled subgroups, which can restrict movement and limit observation of colony-level dynamics. Here, we present a semi-folded arena designed for whole-colony observation of termite movement under controlled conditions. We developed a circular semi-folded arena that remained continuously connected to an intact nest and allowed individuals to move across a central observation surface while recirculating through a folded peripheral section. Using whole colonies of the Neotropical termite Constrictotermes cyphergaster, we recorded exploratory activity under baseline conditions, in the absence of added food or water. High-resolution video recordings were analysed using automated movement extraction to recover trajectories and visualise collective trail structure. Within the first 6 min of activity, collective trail structure was observed in 15 of the 16 colonies analysed. Under these conditions, the semi-folded setup captured early collective trail structure, visible as convergence of cumulative trajectories along shared routes radiating from the arena entrance region. Automated movement extraction was compatible with dense whole-colony recordings and yielded large quantities of positional data during the initial observation interval. Descriptive trajectory-based outputs, including speed distributions for workers and soldiers, showed that the recordings were suitable for recovery of fine-scale movement information. Repeatedly used routes were also often marked by visible dark traces on the paper lining by the end of the observations, providing a qualitative record of cumulative route use. The semi-folded arena provides a practical method for recording whole-colony termite movement under laboratory conditions while maintaining continuous nest access and avoiding manual transfer of individuals during trials. Rather than replacing conventional arena designs, this approach offers an additional methodological option for studying emergent movement patterns in species for which whole-colony observation is feasible. More broadly, it expands the experimental toolkit available for investigating colony-scale spatial organisation under controlled conditions.

Contextualizing the movements of animals into their three-dimensional (3D) habitat contexts is still a major challenge for fields relying on animal tracking methods. This promises to change as Structure-from-Motion photogrammetry tools and techniques revolutionize image processing into the truly 3D spatial realm, by enabling reconstructions of habitat models from overlapping photographs. Combined with tracking data, these techniques would help elucidate drivers behind animal movements that have been previously masked by two-dimensional approaches. Unfortunately, tracking methods are often still impractical for use with understudied or non-model animals, especially those living underwater. In this paper, we describe a method for tracking the translational movements of animals into a photogrammetric habitat model. Our approach spans three general parts: (1) filming the navigation paths of wild animals by following individuals with small cameras (GoPros) on extendable sticks whilst SCUBA diving, (2) reconstructing a 3D spatial habitat model from separate footage, and (3) manually plotting the 3D trajectories of animals into the habitat model. We used one popular commercial software for photogrammetric reconstruction, trajectory plotting, and measurement of trajectories, after which the plots can be exported in a variety of formats for further analyses. Straightforward and flexible methodologies such as this stand to encourage more fieldwork concerning animals that live in structurally complex habitats, or animals that are underrepresented in movement or navigation research. We expect that this approach can be adapted to study many aquatic or terrestrial animals in different habitats, and at various scales.

In certain groups of squids and cuttlefish, males display alternative reproductive tactics (ARTs). In squids, size-associated male dimorphism appears in mating posture, spermatophore transfer site (insemination dimorphism), and sperm flagellar length (sperm dimorphism). Sperm dimorphism is closely linked to insemination dimorphism, in which sperm are deposited either at the females external buccal mass or within her mantle cavity. Insemination dimorphism shapes post-copulatory sperm environments, including the mode of storage, the risk of sperm competition, and the site of fertilization. Therefore, sperm dimorphism is regarded as an adaptive consequence of insemination dimorphism. Conversely, in cuttlefish, both large consorts and small female-mimicking (sneaker) males deposit spermatophores in the same region of the female buccal mass, indicating the absence of insemination dimorphism. In Sepia esculenta and Sepia lycidas, sperm located at the distal end of the male reproductive tract possess longer flagella than those near the testis. This pattern is consistent across all male sizes, suggesting that as males grow, they produce sperm with shorter flagella. Additionally, these within-individual differences in sperm size are greater in smaller males and are positively correlated with relative testis mass. These findings indicate that insemination dimorphism is not required for the evolution of dimorphic sperm in cuttlefish. In both squids and cuttlefish, sperm from smaller males must either enter the female receptacle for extended storage or swim faster to compete with the more abundant consort sperm. These unfavorable conditions associated with the sneaker tactic may drive costly sperm evolution.

The multivoltine non-native box tree moth, Cydalima perspectalis (Lepidoptera: Crambidae), was accidentally introduced in several continents and exhibited a very rapid expansion in its invaded ranges, colonising around 40 countries in less than twenty years. Initially observed in urban areas, it quickly spread to forest stands, causing significant damage to boxwood trees. If the role of the ornamental plant trade in its fast invasion has been investigated, the insects flight capabilities has thus far been overlooked, limiting estimations of its natural dispersal. In this context, we assessed the flight performance of the adults and investigated potential effects of sex, age and generation using computer-monitored flight mills. Under these controled conditions, we estimated the distance flown per night and over the adults lifespan in both sexes for each generation. The adults were able to fly on average 18 km within their lifespan, even though distances flown were highly variable, including several females capable of performing long-distance flights (up to 150 km). The distances covered were partly correlated with the age and body mass of the adults. Mated females flew longer distances than virgin ones, but long dispersal seemed to limit their fecundity. Finally, the overwintering generation presented the highest flight capabilities with individuals able to cover 22 km on average while the late summer generation covered only 10 km. The box tree moth showed good dispersal abilities that likely played a significant role in its rapid local expansion, while human-mediated dispersal favoured its long-distance dispersal.

The plasmodium of Myxogastoria (a group of amoeboid protists) species often crawls around the forest floor to feed while searching for places to form fruiting bodies for reproduction (sporulation). Certain environmental factors that trigger sporulation have been reported; however, other unknown factors are also expected. In this study, we reported field observations of Physarum rigidum and Fuligo septica. Inspired by the field observation, we examined the effects of multiple factors on sporulation in laboratory experiments using Physarum polycephalum. We found that:(1) there was a critical body size below which sporulation did not occur under our experimental conditions and (2) the plasmodium selected its sporulation sites from the available landscape of the experimental arena: dry and low sites for the majority and dry and high sites for the minority. Further analysis revealed that they preferred the edge area at the high site. We discuss the possible ecological importance of the threshold and location preference

For oviparous animals, the decision of where to lay eggs is critical, as offspring remain sessile from oviposition through hatching and are thus unable to escape unfavorable conditions. Consequently, females are expected to select oviposition sites that benefit embryo development and survival. This may be particularly relevant for arboreal frogs, which typically lay eggs on leaves overhanging water, where embryos are exposed to predation, desiccation, and other risks until hatching. Yet studies directly linking maternal substrate choice to embryo survival remain scarce. Here, we examine how oviposition substrate influences embryo survival in the Emerald glass frog (Espadarana prosoblepon), a species in which females deposit eggs on multiple substrates, providing a rare opportunity to test how oviposition decisions affect reproductive success. Monitoring clutches in situ, we compared microclimatic conditions, hatching success, and sources of embryo mortality between the most used substrates: the spike moss Selaginella diffusa and leaves. Additionally, we conducted a two-choice experiment in semi-captivity to test whether females preferentially select one substrate over the other. Although microclimatic conditions did not differ between substrates, hatching success was significantly higher on S. diffusa, which also experienced less predation. In the two-choice experiment, all females laid their eggs on S. diffusa, and those clutches had higher hatching success and faster embryonic development rates than those on leaves. Together, these results support the hypothesis that non-random oviposition site selection in E. prosoblepon is driven by the maximization of embryo survival, demonstrating that substrate choice has measurable fitness consequences for the offspring.

Predators are often considered regulators of disease in prey populations, a concept central to the "healthy herd hypothesis". This hypothesis suggests that by preferentially removing infected individuals, predators can reduce parasite prevalence. However, predators may also act as disease vectors, facilitating the spread of parasites. We investigated whether stickleback fish (Gasterosteus aculeatus) can act as vectors for the transmission of the obligate bacterial parasite Pasteuria ramosa to its Daphnia host, a widespread freshwater zooplanktor. We fed infected D. magna to sticklebacks, and subsequently analysed faecal samples for the presence, viability, and infectivity of parasite transmission stages (= spores). We recovered approximately 60% of the consumed spores from fish faeces and these spores did not suffer from reduced infectivity to D. magna. Additionally, spores associated with sloppy feeding did not reduce infection rates. Thus, consumption of infected hosts by fish does not eliminate the parasite, but in contrary, may contribute to the spread and persistence of P. ramosa in natural populations, potentially influencing parasite dynamics in natural freshwater ecosystems.

Globally, nocturnal lightscapes are now determined by both moonlight and light pollution, or artificial light at night (ALAN). Organisms respond both to changes in moonlight across lunar cycles and to alterations in light conditions due to artificial light. The interaction of natural and artificial light is a critical aspect to incorporate into our understanding of how crepuscular and nocturnal ecology is altered in anthropogenically-modified landscapes. In this manuscript we review the rapidly expanding body of research on ecological impacts of ALAN to (1) assess patterns of lunar data inclusion and (2) summarize documented interactions of moonlight and ALAN. Three-fourths (72%) of 379 papers reviewed did not incorporate moonlight into their statistical analyses and experimental design. Only 12% directly investigated interaction effects of moonlight and ALAN. However, 70% of these studies reported an interactive effect. Considering this stark contrast, as a precursor to our literature review, we present an overview of moonlight and the lunar cycle for biologists. The overarching trend emerging from the literature is that biological impacts of ALAN decrease with increasing moonlight, although the opposite is true in some cases. After summarizing the literature, we present general hypotheses regarding the interaction of the lunar cycle and ALAN. These hypotheses consider the different forms of ALAN encountered by organisms (i.e. skyglow and light sources) and account for the influence of cloud cover. Finally, we suggest best practices for incorporating moonlight into future research on biological impacts of ALAN.