Journal of Comparative Physiology A

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.

Understanding the nutritional preferences of honey bees (Apis mellifera) is essential for comprehending their behavioral ecology and the division of labor within a colony. While gustatory sensitivity to sucrose is well-documented in workers, a significant research gap exists regarding the sensory responses of queens and their reactions to caste-specific nutrition such as royal jelly. This study utilized the proboscis extension response (PER) assay to compare the food preferences of three distinct bee categories: foragers, 1-day-old workers, and queens. Subjects were presented repeatedly, in a pseudorandom order, with water, sucrose, royal jelly, and a sucrose-royal jelly mixture as gustatory stimuli. Foragers exhibited a high responsiveness to sucrose and showed uniformly low responsiveness to other stimuli. Although 1-day-old workers showed high responsiveness to sucrose, unlike foragers, they also responded to the sucrose-royal jelly mixture. Queens displayed a unique response profile, with near-ceiling responsiveness to both royal jelly and the mixture, followed by response to sucrose solution without habituation. Additionally, responsiveness to the sucrose was higher in foragers than in 1-day-old workers. These findings suggest that the honey bee gustatory and sensory system is tuned to the specific nutritional requirements of caste and age.

Tropical pollinators forage in environments where floral resources vary in space and time, requiring flexible strategies to optimise foraging efficiency. One such strategy, floral constancy - the temporary restriction to a single flower type - strongly influences foraging success and plant-pollinator interactions. We aimed to: (1) quantify spontaneous colour preferences and constancy in the Asian giant honeybee Apis dorsata, (2) test whether reward concentration modulates these preferences, (3) evaluate how quickly learned associations override spontaneous biases, (4) determine whether bees can use multiple colour associations simultaneously, and (5) assess whether local floral spectral patterns correlate with bee preferences. Bees trained to a neutral UV-grey stimulus showed a strong spontaneous preference and high constancy to blue, revealing a robust short-wavelength bias. Crucially, the strength of this spontaneous bias depended on reward concentration; Low-reward conditions elicited strong blue constancy, whereas high-reward conditions weakened it, demonstrating that reward expectation shapes spontaneous colour choices. This bias was flexible. When bees learned that yellow was rewarding they switched their preferences. Bees sequentially trained to both colours visited blue and yellow, showing no overall bias, or effect of the last-trained colour, indicating that recent experiences disrupt colour-specific constancy and generate largely random foraging choices. Bees were capable of learning and retaining two colours simultaneously, effectively suppressing the influence of spontaneous preferences. Finally, analysis of the communitys floral spectral distribution revealed a strong dominance of short-wavelength flowers, suggesting that long-term selection by the local floral environment may underlie the spontaneous blue preference observed in A. dorsata.

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.

Nociception is an essential response for organisms to avoid potential harm and promote survival. Its molecular determinants are largely conserved across Eumetazoa. TRPV receptors are polymodal ion channels exhibiting selective peripheral expression and functional coupling that underpins nociception and pain modulation in complex organisms. However, the execution of protective behaviours triggered by TRPVs is also found in species with a simpler nervous organisation, thus encouraging their investigation in invertebrate model organisms to increase understanding of animal nociception. Cephalopods represent an interesting invertebrate phylum with respect to the evolution of the nervous system, whose complexity suggests it might support pain-like states that exist in vertebrates. This possibility is reflected by the inclusion of cephalopods in the UK and EU animal welfare legislations. Despite this, there is poor characterisation of cephalopod molecular nociceptors. For this reason, we used in silico analysis to identify two TRPV channels in Octopus vulgaris genome (Ovtrpv1 and Ovtrpv2). We validated the putative transcript sequences and highlighted prevalent expression in sensory tissues. We investigated the functional competence of these TRPVs by heterologously expressing Ovtrpv1 and Ovtrpv2 cDNA into Caenorhabditis elegans null mutants of the orthologous genes, ocr-2 and osm-9 respectively. Ovtrpvs successfully rescued the aversive response to chemical and mechanical noxious stimuli in the C. elegans mutants, suggesting these receptors are polymodal nociceptors. Additionally, complementary investigation using Xenopus laevis oocytes showed Ovtrpv1 and Ovtrpv2 form an active heteromeric channel gated by nicotinamide. This study highlights Ovtrpvs as an important route to better understand nociceptive detection in cephalopods.

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

Animals need to precisely perceive and integrate the environmental cues to orient and select the appropriate motor responses required for navigating. This is the case, for instance, of the optokinetic reflex (OKR) and the optomotor response (OMR) in Drosophila melanogaster, where optic flow stimulation modulates the head or the body and legs motor activity respectively. Despite large bodies of literature on both the OKR and the OMR, there is still a limited understanding, in flies, of the impact on these responses of concomitant, and potentially conflicting, sensory inputs. To investigate this aspect, we used fruit flies walking on a sphere, presented with optic flow stimulation leading to the OMR together with the simultaneous exposure to olfactory stimulation, either using established repellent or masking compounds. We analysed the effect of different substances, and of their concentration, on the dynamics of the flies response to moving gratings, evaluating the fly walking path as well as average speed and duration. This analysis revealed several alterations between the compounds tested, in agreement with reported data on the simpler OKR. In conclusion, we show that concomitant exposure to repellents and maskers may consistently affect fundamental processes (the OKR and OMR) available to insects for informing themselves while navigating through the environment.

Organisms are exposed to predictable daily and seasonal environmental oscillations. Biological clocks enable organisms to anticipate these changes and coordinate physiology and behaviour accordingly. While circadian mechanisms are well studied in terrestrial model organisms, little is known about the neuronal organisation of biological clocks in ecologically important species, especially in the marine environment. Antarctic krill (Euphausia superba) is central to the functioning of the Southern Ocean ecosystem and relies on precise timing to cope with the extreme, high-latitude fluctuations in photoperiod, food availability, and sea-ice cover in its habitat. Despite evidence for circadian and seasonal rhythms in krill behaviour and physiology, the neuronal architecture underlying these timing processes has remained unresolved. In this study, we use in situ hybridisation and antibody staining to characterise the circadian clock in the krill brain. Immunostaining with an antibody against crustacean {beta}-Pigment-dispersing hormone ({beta}-PDH) reveals distinct clusters of PDH-positive neurons in the optic lobes and dorsal central brain, along with an extensive PDH-positive fibre network. We further localise transcripts of the core clock genes cryptochrome-2 (cry2) and period (per) in cell clusters in the optic lobes, which also include the PDH-positive neurons. More specifically, PDH-positive neurons are a subgroup of the cry2 and per-positive cells. Together, these findings provide the first description of the neuronal architecture of the circadian clock in Antarctic krill and establish essential groundwork for future studies on biological timing, environmental adaptation, and the resilience of this key species in a rapidly changing Southern Ocean.

Astyanax mexicanus consists of eyed, river-dwelling "surface" fish, and multiple, independently evolved cave populations, which have converged on troglobitic traits such as eye loss and reduced metabolism. However, considerably less is known about constructive adaptations, which include a larger olfactory epithelium in cavefish. It is unknown how this relates to the olfactory bulb (OB), which is the first stage of olfactory sensory processing in the brain. The goal of the present study is to begin to define the structure and functional organization of the OB in A. mexicanus, and to begin to understand how it was transformed via cave adaptation. We addressed these questions using whole-mount immunohistochemistry and in vivo Ca2+ imaging from the OB of developmentally matched surface and Pachon cavefish. The cavefish OB was significantly larger than surface fish by 14 days post fertilization (dpf), which was accompanied by a broad and proportional increase in synaptic input to most glomerular regions. Increases in the size of the OB were accompanied by increases in the number of neurons expressing tyrosine hydroxylase and calretinin, the latter of which occurred primarily in the medial OB and could not be explained as a compensatory response to a larger OB. In vivo Ca2+ imaging from the dorsal OB of surface and cavefish in response to a panel of chemical stimuli revealed odor-evoked responses that were spatially organized and highly conserved across the two populations. Surprisingly, the medial OB was consistently activated by any change in water flow in both populations, although the number of water-responsive neurons was significantly greater in cavefish when measurements were performed using either in vivo imaging or the neuronal activity marker phospho-ERK. Water-responding neurons were similarly present in the olfactory epithelium in both populations, along with neurons expressing the mechanosensitive ion channel Piezo2, with significantly more Piezo2-expressing neurons present in cavefish. Therefore, cavefish exhibit enhanced multisensory integration of olfactory and mechanosensory input in the earliest stage of olfactory sensory processing in the brain.

Communication allows organisms to quickly convey information vital for survival or fitness. Chemical communication and speed-accuracy trade-offs are ubiquitous in animal decision making. Most studies have used species which forage mainly above-ground species, tested in an epigean setting, but it remains unclear whether below-ground species behave similarly. Here, we use the below-ground ant Tetramorium alpestre to assess the efficacy of above- vs. below-ground mazes, the accuracy of decisions when using natural vs. artificial pheromones, the presence of a speed-accuracy trade-off, and the pheromones effect on aggression. Ants decided more quickly under below-ground than above-ground conditions, indicating they may be distracted by above-ground stimuli. Ants followed both natural and artificial trails but in direct competition preferred artificial trails, likely due to a higher pheromone concentration. Surprisingly, no speed-accuracy trade off was observed during path decision-making. Lastly, population origin but not pheromones affected if and how aggression occurred in presence of trail and home-range marking pheromones. We argue that the design of behavioural tests should match the lifestyle of the focal organism. We further speculate that speed-accuracy trade-offs likely are highly species- and context-specific and other factors besides chemicals seem important to trigger aggression, at least in this species.

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.

The rat vestibular system plays a critical role in anti-gravity responses such as the tail-lift reflex and the air-righting reflex. In a previous study in male rats, we obtained evidence that these two reflexes depend on the function of non-identical populations of vestibular sensory hair cells (HC). Here, we caused graded lesions in the vestibular system of female rats by exposing the animals to several different doses of an ototoxic chemical, 3,3-iminodipropionitrile (IDPN). After exposure, we assessed the anti-gravity responses of the rats and then assessed the loss of type I HC (HCI) and type II HC (HCII) in the central and peripheral regions of the crista, utricle and saccule. As expected, we recorded a dose-dependent loss of vestibular function and loss of HCs. The relationship between hair cell loss and functional loss was examined using non-linear models fitted by orthogonal distance regression. The results indicated that both the tail-lift reflex and the air-righting reflexes mostly depend on HCI function. However, a different dependency was found on the epithelium triggering the reflex: while the tail-lift response is sensitive to loss of crista and/or utricle HCIs, the air-righting response rather depends on utricular and/or saccular integrity.

In natural environments, animals must effectively maneuver around obstacles to reach goals such as food or shelter. Recent work has demonstrated that laboratory mice use vision for naturalistic behavior such as prey capture, escape, and distance estimation. However, it is unknown to what extent mice use vision relative to other senses such as touch for obstacle avoidance, a critical natural behavior. In this study we developed an obstacle avoidance task in freely moving mice to investigate how vision is used to guide paths around an obstacle obstructing a goal. We found that mice clearly use vision to avoid an obstacle, steering around the obstacle at distances where tactile information isnt available. By comparing trajectories for mice performing obstacle avoidance in the light versus the dark, we found that vision contributes to more spatially efficient trajectories and paths directed to the open edge of the obstacle. When vision is available, mice make large orienting movements towards the opening of the obstacle at about 10 cm from its edge, suggesting that mice are actively using visual information to direct these movements. Finally, by occluding one eye, we found that mice were still able to avoid obstacles with primarily monocular information. Taken together, these results demonstrate that laboratory mice use vision to avoid an obstacle, taking directed paths that are initiated by large orienting movements. In addition to demonstrating the visual behavioral capabilities of the mouse, this paradigm can serve as a foundation to study the neural circuits that mediate visually guided orienting and locomotion. HighlightsO_LIWe developed a simple obstacle avoidance task for freely moving mice that requires minimal training C_LIO_LIVision is necessary for efficient and directed paths around an obstacle C_LIO_LIMice steer around obstacles by performing directed head movements towards clear paths C_LIO_LIMice do not require binocular vision for obstacle avoidance C_LI

Foraging bee decision-making research has focused on choice determinants, and the variability and underlying causes of pre-choice latency remain understudied. Here, individual bumblebees (Bombus impatiens) were trained to associate one colored stimulus with a medium-value reward and another with a novel, higher-value reward. The experimental design consists of seven blocks, each containing four consecutive single-stimulus presentations followed by a forced binary choice. The latency to choose a stimulus and the type of choice during dual-choice trials were analysed. In dual-choice trials, bees in the yellow-high reward group showed a slower increase in high-reward selection than those in the blue-high group, suggesting persistent innate color bias. Response latencies for the low-reward stimulus systematically increased across blocks, indicating progressive devaluation. Early learning phases showed a temporary increase in response latency, extending previous findings on experience-dependent adjustments in acceptance thresholds. Latency in single-stimulus trials correlated with binary choice results, though choice proved a stronger indicator of preference than latency. Certain options elicited faster responses when presented with an alternative than when presented alone. Together, these findings support a deliberative model of bumblebee decision-making, in which pre-choice latency is modulated by innate preferences, associative learning, and immediate context.

The ability of animals to innovate - solve novel problems - can shape their ecology and evolution. Here we investigate how individual traits and environmental complexity relate to successful solving of a novel problem. We presented foraging bumble bees (Bombus impatiens) with artificial flowers of not-previously-encountered shapes and recorded the bees latency to access nectar. We measured individual foraging traits across multiple trips with simple flowers that did not require innovation, and bees were foraging either in a simple or complex environment (cluttered flight arena). Bees in complex environments took longer to find and were less likely to land on novel flowers, indicating that environmental complexity may take up cognitive resources and make search more difficult. However, we did not find an effect of environmental treatment on the ability or time to access reward in novel flowers once bees had landed on them. In contrast, behavioral traits significantly predicted how quickly bees solved novel flowers. In particular, overall foraging tempo as well as routine formation, i.e. how much bees followed a fixed route on known flowers, predicted innovation - faster bees innovated faster, and bees with more repetitive foraging sequences were slower to solve the novel tasks. Overall, while the degree of evolutionary novelty in tasks or solutions is always hard to evaluate, our findings demonstrate that environment and individual traits may affect innovation in different ways. Individuals in simple environments may be more likely to detect, and individuals that are generally faster and have a lower tendency to develop fixed routines may be more likely to solve, novel tasks.

Injuries are common in animals and represent a major threat to individual survival. They can result from inter- or intraspecific conflict, predation, or pugnacious prey. Despite their potential ecological and evolutionary importance, injury patterns remain poorly documented in animal populations. To test whether a species feeding ecology or habitat can predict injury patterns, we quantified injury rates and affected body regions among native ant species collected from different habitats in Bavaria, Germany. Specimens were sampled using pitfall traps, which proved to be an efficient method for injury assessment. Injury rates varied substantially among species and genera, ranging from 0% to 38%. Predatory ant species exhibited higher frequencies of leg injuries, whereas omnivorous species were more frequently injured at the antennae. The distribution of injuries likely reflects both foraging ecology and species-specific wound care behaviors, with a high frequency of trochanter injuries potentially indicating prior amputation events to cope with infected leg injuries, as observed in Lasius alienus. Our findings demonstrate that injury propensity and distribution are shaped by feeding habits and behavioral adaptations, providing comparative evidence that the costs and management of injuries vary systematically among ant species. Our study thus highlights injuries as a measurable axis of selection that may have contributed to the emergence of wound care and other forms of social immunity in ants.

Climate warming alters the thermal environment experienced by ectotherms, whose physiological performance and fitness are constrained by temperature. Early life stages are often the temperature-sensitive phases of the life cycle, with potential consequences for population persistence, particularly in freshwater stenotherms such as the Arctic charr (Salvelinus alpinus). The persistence of populations will partly depend on the adaptive potential of critical life stages to environmental changes. In this study, we used a common garden approach to compare the response and phenotypic plasticity of four charr populations to warmer conditions. These populations inhabit thermally contrasted lakes and differ in origin (native/introduced) and management history. We reared embryos at either an optimal (5{degrees}C) temperature for larval development or a warmer but realistic (8.5 {degrees}C) temperature. We tested adaptive divergence among populations in four traits (survival, incubation duration, body length and yolk sac volume), using Qst - Fst comparisons. We report negative effects of temperature on body size, survival and earlier hatching. Thermal reaction norms differed among populations, indicating adaptive divergence. Contrary to expectations, populations originating from warmer environments did not consistently exhibit higher trait values under elevated temperatures. In contrast, the unmanaged and colder high-altitude population exhibited higher survival rates and lower yolk reserves for a given size under heat stress than the other populations. Our results suggested that evolutionary trajectories specific to each population are shaped by factors related to the populations history, including introductions, demographic fluctuations and long-term repopulation practices, which can jointly influence the potential for adaptation to heat stress.

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.

The growth of bivalve shells is neither homogeneous nor continuous in time, resulting in the formation of growth patterns that correspond to the alternation of growth lines and increments deposited at regular intervals of time. The control of periodic increment formation is poorly understood and several hypotheses have been proposed. It has been proposed that environmental factors directly impact shell growth patterns, although it occasionally fails to adequately explain the observed shell growth patterns. The present study investigates the alternative hypothesis that the process of shell biomineralisation is controlled by biological clocks. This study demonstrates the existence of a functional circadian clock in M. galloprovincialis, as evidenced by molecular and behavioural results. Core circadian clock genes and biomineralisation genes have been observed to be expressed in the same cells of the mantle as revealed by in situ hybridisation experiments. However, the expression of core circadian clock genes and biomineralisation genes tested in situ and in aquaria exhibited different rhythmic profiles. This finding suggests that the clock does not directly activate the expression of the targeted biomineralisation genes in the mantle. Nevertheless, a significant rhythm of expression of biomineralisation-related genes was observed in mussels reared under free-running conditions, revealing the endogenous nature of the rhythm. The present study suggests that biological clocks play a role in controlling shell biomineralisation in M. galloprovincialis, although the precise underlying mechanism remains to be elucidated.

When novel nutrient-rich food sources become available to species sharing the same natural habitat, interspecies competition may arise, yielding insights into the ecological and social dynamics of the observed species. Here, we investigated food consumption patterns, and consequent social interactions, by two sympatric species of mice in response to a novel nutrient-rich food source. By deploying, in the mices natural habitat, baited video-monitored chambers, we collected, over a 5-month period, 1805 observations of food visiting by Apodemus agrarius and Apodemus flavicollis. We also documented interspecific encounters, with 86.7% of the cases showing agonism. In these interspecies agonistic encounters, A. flavicollis was always the initiator of agonism, attacking within 2 sec in 92.3% of the cases, and being dominant over A. agrarius in 84.6%. Analysis of food visiting behavior revealed that, initially, both species preferred nocturnality. However, after the interspecies fights, A. agrarius switched its temporal preference to diurnality, leading to temporal niche segregation between the two species and a significant reduction of interspecies encounters. Moreover, A. agrarius demonstrated hour-specific avoidance of A. flavicollis, visiting significantly less in hours with A. flavicollis compared to hours without. Through temporal niche switching, A. agrarius managed to access the food source safely, without fights. In contrast, A. flavicollis remained consistently nocturnal across the entire study. Notably, our study presents the first 24h foraging actogram for free-living rodents. Moreover, while rodent interspecific competition is a well-known phenomenon, most of what we know about it comes from indirect observations. Direct observations of rodent interspecific interactions in nature are rare. Our work is the first direct (video-monitored) observation of temporal switch-inducing interspecies interactions in nature. As free-living rodents are currently considered a major model system for the study of interspecific competition, these results may offer precious insights for a better understanding of social dynamics, especially in asymmetric relationships. Furthermore, our findings highlight the significance of considering temporal dynamics in studies of interspecific interactions.