Biology Letters

Cortical neurons that make direct connections to motor neurons in the brainstem and spinal cord are specialized for fine motor control and learning [1, 2]. Imitative vocal learning, the basis for human speech, requires the precise control of the larynx muscles [3]. While much knowledge on vocal learning systems has been gained from studying songbirds [4], an accessible laboratory model for mammalian vocal learning is highly desirable. Evidence indicative of complex vocal repertoires and dialects suggests that bats are vocal learners [5, 6], however the circuitry that underlies vocal control and learning in bats is largely unknown. A key feature of vocal learning animals is a direct cortical projection to the brainstem motor neurons that innervate the vocal organ [7]. A recent study [8] described a direct connection from the primary motor cortex to medullary nucleus ambiguus in the Egyptian fruit bat (Rousettus aegyptiacus). Here we show that a distantly related bat, Sebas short-tailed bat (Carollia perspicillata) also possesses a direct projection from the primary motor cortex to nucleus ambiguus. Our results, in combination with Wirthlin et al. [8], suggest that multiple bat lineages possess the anatomical substrate for cortical control of vocal output. We propose that bats would be an informative mammalian model for vocal learning studies to better understand the genetics and circuitry involved in human vocal communication.

Male animals are not given equal mating opportunities under competitive circumstances. Small males often exhibit alternative mating behaviours and produce spermatozoa of higher quality to compensate for their lower chances of winning physical contests against larger competitors [1]. Because the reproductive benefits of these phenotypes depend on social status/agonistic ranks that can change during growth or aging [2], sperm traits should be developed/switched into fitness optima according to their prospects. However, reproductive success largely relies upon social contexts arising instantaneously from intra- and inter-sexual interactions, which deter males from developing extreme traits and instead favour behavioural plasticity. Nevertheless, the extent to which such plasticity influences developmentally regulated alternative sperm traits remains unexplored. Squids of the family Loliginidae are excellent models to investigate this, because they show sophisticated alternative reproductive tactics (ARTs) by which small males, known as "sneakers", produce longer spermatozoa and perform extra-pair copulation to attach their sperm packages near the female seminal receptacle (SR). In contrast, large "consort" males have shorter spermatozoa and copulate via pair-bonding to insert their sperm packages near the internal female oviduct [3]. In addition, plasticity in male mating behaviour is common in some species while it is either rare or absent in others. Thus, squid ARTs display a broad spectrum of adaptive traits with a complex repertoire in behaviour, morphology and physiology [3].

The nearly constant downward force of gravity has powerfully shaped the behaviors of many organisms [1]. Walking flies readily orient against gravity in a behavior termed negative gravitaxis. In Drosophila this behavior is studied by observing the position of flies in vials [2-4] or simple mazes [5-9]. These assays have been used to conduct forward-genetic screens [5, 6, 8] and as simple tests of locomotion deficits [10-12]. Despite this long history of investigation, the sensory basis of gravitaxis is largely unknown [1]. Recent studies have implicated the antennae as a major mechanosensory input [3, 4], but many details remain unclear. Fly orientation behavior is expected to depend on the direction and amplitude of the gravitational pull, but little is known about the sensitivity of flies to these features of the environment. Here we directly measure the gravity-dependent orientation behavior of flies walking on an adjustable tilted platform, that is inspired by previous insect studies [13-16]. In this arena, flies can freely orient with respect to gravity. Our findings indicate that flies are exquisitely sensitive to the direction of gravitys pull. Surprisingly, this orientation behavior does not require antennal mechanosensory input, suggesting that other sensory structures must be involved.

Bats exhibit a diverse array of social behaviors, yet detailed studies on the intricacies of these interactions, particularly in rare species like the spectral bat (Vampyrum spectrum), remain scarce. This study presents the first comprehensive description of prey sharing and other social behaviors in a wild social group of V. spectrum. Over several months, we conducted extensive video recordings in a hollow tree, used as a day- and night-roost, to document these behaviors, aiming to elucidate key social interactions among the bats and their role within their ecological niche. We observed various remarkable social behaviors, including food sharing between family members. Our findings support the hypothesis that prey sharing as a form of biparental care may serve as a method for adults to transition young bats from milk to a carnivorous diet, ensuring adequate food intake and allowing them to practice how to handle large prey items. We also challenge the notion that V. spectrum forages exclusively solitarily, as we documented several instances of synchronized roost departures and returns and thus presumably cooperative foraging. This indicates a more complex social structure and behavioral ecology than previously understood. Our comprehensive analysis of observational data enhances our understanding of the social dynamics of V. spectrum, providing new insights into the evolution of cooperative behaviors in bats. HighlightsExtensive parental care: Most likely both male and female spectral bats participate in parental care, e.g. food provisioning, a rare behavior among mammals Prey sharing as a social cooperative behavior: Detailed observations of prey sharing within the roost, highlighting social cooperation and resource allocation Extended dependency period for pups: A prolonged developmental period allows pups to receive care and acquire essential survival skills until they achieve full independence Juvenile play: Pups engaged predominantly in social play (mock fights), but also played with objects or food items, a rare behavior among bats

Two genera and multiple species of short-faced bear from the Americas went extinct during or toward the end of the Pleistocene, and all belonged to the endemic New World subfamily Tremarctinae [1-7]. Two of these species were giants, growing in excess of 1,000 kg [6, 8, 9], but it remains uncertain how these extinct bears were related to the sole surviving short-faced bear: the spectacled bear (Tremarctos ornatus). Ancient mitochondrial DNA has recently suggested phylogenetic relationships among these lineages that conflict with interpretations based on morphology [1, 10-12]. However, widespread hybridisation and incomplete lineage sorting among extant bears mean that the mitochondrial phylogeny frequently does not reflect the true species tree [13, 14]. Here we present ancient nuclear genome sequences from representatives of the two extinct short-faced bear genera, Arctotherium and Arctodus. Our new data support a third hypothesis for the relationships among short-faced bears, which conflicts with existing mitochondrial and morphological data. Based on genome-wide D-statistics, we suggest that the extant spectacled bear derives substantial ancestry from Pleistocene hybridisation with an extinct short-faced bear lineage, resulting in a discordant phylogenetic signal between the mitochondrion and portions of the nuclear genome.

Vocalisations play a crucial role in the social systems of many animals, and may inadvertently reveal behavioural characteristics of the sender. Bats, the second largest mammalian order, rely extensively on vocalisations due to their nocturnal lifestyle and complex social systems, making them ideal for studying links between vocalisations and consistent behavioural traits. In this study, we developed a new testing regime to investigate if consistent individual vocalisation differences in nectarivorous bats are associated with specific behavioural types. We exposed 60 wild, male Glossophaga soricina handleyi bats to novel and risky stressors, and assessed their behavioural and vocal responses. Proactive, exploratory, and bold bats were more likely to produce social calls, and among the vocalising bats, more agitated bats produced higher numbers of social calls. We thus show that bat vocalisation behaviour can be indicative of a certain behavioural type, potentially allowing conspecifics to assess personalities from a distance, which in turn could impact subsequent social interactions, group dynamics, and reproductive success. Our results, in combination with previous findings in birds, suggest that advertent or inadvertent long-distance broadcasting of personality may be widespread, thus opening up new exciting questions about the links between vocalisations and sociality.

Individual recognition is conceptually complex and computationally intense, leading to the general assumption that this social knowledge is solely present in vertebrates with larger brains, while miniature-brained animals in differentiating societies eschew the evolutionary pressure for individual recognition by evolving computationally less demanding class-level recognition, such as kin, social rank, or mate recognition. Arguably, this social knowledge is restricted to species with a degree of sociality (sensu [1], for a review [2]). Here we show the exception to this rule in a non-social arthropod species, the jumping spider Phidippus regius. Using a habituation - dishabituation paradigm, we visually confronted pairs of spatially separated spiders with each other and measured the interest of one spider towards the other. The spiders exhibited high interest upon initial encounter of an individual, reflected in mutual approach behaviour, but adapted towards that individual when it reoccurred in the subsequent trial, indicated by their preference of staying farther apart. In contrast, spiders exhibited a rebound from habituation, reflected in mutual approach behaviour, when a different individual occurred in the subsequent trial, indicating the ability to tell apart spiders identities. These results suggest that P. regius is capable of individual recognition based on long-term memory.

Neural activity in mouse primary visual cortex (V1) correlates strongly with orofacial movements. Such strong modulation has not been found in the primate visual cortex during eye fixation [1], which led to the suggestion that the modulation may primarily depend on eye movements in both species [2]. Here we examined the influence of eye movements on neural activity in mouse visual cortex both in complete darkness and in the presence of different types of visual input. In all cases, we found that eye movements explain a smaller fraction of neural activity variance compared to orofacial behaviors. Additionally, we found that eye movements were correlated to orofacial movements, such as whisking and sniffing, and thus may be indirectly correlated to neural activity. These results further emphasize the impact of movement signals on mouse visual cortex during free viewing behavior.

Stinging ants have diversified into various ecological niches, and several evolutionary drivers may have contributed to shape the composition of their venom. To comprehend the drivers underlying venom variation in ants, we selected 15 Neotropical species and recorded a range of traits, including ecology, morphology, and venom bioactivity. Principal component analysis of both morphological and venom bioactivity traits revealed that stinging ants display two functional strategies. Additionally, phylogenetic comparative analysis indicated that venom function (predatory, defensive, or both) and mandible morphology significantly correlate with venom bioactivity and amount, while pain-inducing activity trades off with insect paralysis. Further analysis of the venom biochemistry of the 15 species revealed switches between cytotoxic and neurotoxic venom compositions in some species. This study highlights the fact that ant venoms are not homogenous, and for some species, there are major shifts in venom composition associated with the diversification of venom ecological functions. SignificanceVenoms are under severe evolutionary pressures, exerted either on the innovation of toxins or the reduction of the metabolic cost of production (1). To reduce the metabolic costs associated with venom secretion, some venomous animals can regulate venom expenditure by metering the amount of venom injected and by switching between offensive and defensive compositions (2-2). Many ants use venom for subduing a wide range of arthropod prey, as well as for defensive purposes against invertebrates and vertebrates, but are unable to adapt venom composition to stimuli (5, 6). Consequently, the expression of venom genes directly affects the ability of ants to interact with the biotic environment, and the venom composition may be fine-tuned to the ecology of each species. A previous study showed that defensive traits in ants exhibit an evolutionary trade-off in which the presence of a sting is negatively correlated with several other defensive traits, further supporting that trade-offs in defensive traits significantly constrain trait evolution and influence species diversification in ants (7). However, the sting is not used for the same purpose depending on the ant species. Our study supports an evolutionary trade-off between the ability of venom to deter vertebrates and to paralyze insects which are correlated with different life history strategies among Formicidae.

Sensory processing of environmental stimuli during locomotion is critical for the successful execution of goal-directed behaviors and navigating around obstacles. The outcome of these sensorimotor processes can be challenged by head movements that perturb the sensory coordinate frames directing behaviors. In the case of visually-guided behaviors, visual gaze stabilization results from the integrated activity of the vestibuloocular reflex and motor efference copy originating within circuits driving locomotor behavior. A recent videographic study showed that echolocating bats exhibit inflight head stabilization during a target identification and landing task, though compensatory timing of the bats sonar signals was not reported. In the present investigation we tested hypotheses that head stabilization is more broadly implemented during epochs of exploratory flight, and is temporally associated with emitted sonar signals, which would optimize acoustic gaze. This was achieved by measuring head and body kinematics with motion sensors secured to the head and body of free-flying Egyptian fruit bats. These devices were integrated with ultrasonic microphones to record the bats sonar emissions and elucidate their temporal association with periods of head stabilization. Head accelerations in the Earth-vertical axis were asymmetric with respect to wing downbeat and upbeat relative to body accelerations. This indicated that inflight head and body accelerations were uncoupled, outcomes consistent with the implementation of head movements that limit vertical acceleration during wing downbeat. Furthermore, sonar emissions during stable flight occurred most often during wing downbeat and head stabilization, supporting the conclusion that head stabilization behavior optimized sonar gaze and environmental interrogation via echolocation. Summary statementDirect measurements of head and body kinematics from affixed motion sensors revealed head stabilization behaviors during exploratory flights in bats. Most sonar emissions were temporally correlated with this behavior, thereby contributing to the optimization of acoustic gaze.

Blood-feeding insects, such as the mosquito, Aedes (Ae.) aegypti, use multiple senses to seek out and bite humans [1, 2]. Upon exposure to CO2, the attention of female mosquitoes to potential human targets is greatly increased. Female mosquitoes use vision to assist them in honing in on hosts that may be up to 10 meters away [3-9]. Only after coming into close range do convective heat from skin and odors from volatile organic compounds come into play, allowing female mosquitoes to evaluate whether the object of interest might be a host [10, 11]. Here, using CRISPR/Cas9 we mutated the gene encoding Op1, which is the most abundant of the five rhodopsins expressed in the compound eyes of Ae. aegypti. Using a cage assay and a wind tunnel assay, we surprisingly found that elimination of op1 did not impair CO2-induced target seeking. We then mutated op2, which encodes the rhodopsin most similar to Op1, and also found that there was no impact on this behavior. Rather, mutation of both op1 and op2 was required to abolish vision-guided target recognition. In contrast to this defect, the double mutants still exhibited normal light attraction. By measuring the optomotor response, we found that the double mutants still recognized moving cues in their environment. In further support of the conclusion that the double mutant is not blind, we found that the animals retained an electrophysiological response to light, although it was diminished. This represents the first perturbation of vision in mosquitoes and indicates that hostseeking by Ae. aegypti depends on redundant rhodopsins.

The volcanic eruption of Toba in northern Sumatra at 71 kyBP ({+/-}5 kyBP) emitted sulfur gas and deposited thick layers of dust throughout the surrounding region. It is thought to have had a significant and dramatic cooling impact on the paleoclimate worldwide. Ambrose [1] conjectured this to be the cause of the contemporaneous (50-100 kyBP) population bottleneck observed in humans. We hypothesize that a volcanic winter of sufficient magnitude to cause a population bottleneck in humans would similarly affect other mammals. To test this hypothesis, we estimated pairwise mismatch distributions using mtDNA control region sequences of 28 mammal species archived on NCBI to assess whether each species underwent a population bottleneck. For any species fitting the sudden expansion model, we estimated the timing of the bottleneck and compared it to the date range of the Toba eruption. Only 3 of the 28 species show evidence of rapid population expansion overlapping in time with the Toba eruption. Therefore, the hypothesis that the volcanic winter triggered by the Toba eruption caused a significant bottleneck impacting mammal species worldwide is not supported by mitochondrial evidence. Our results question the hypothesis that the Toba eruption contributed to the bottleneck observed in humans at this time.

There are immense consistent inter-individual differences in animal behavior. While many studies have documented such behavioral differences, often referred to as individual personalities, little research has focused on the underlying causes and on determining whether they are innate or based on individual experience. Moreover, most studies on animal personalities have described consistent differences in behavior under laboratory conditions. We aimed to examine the impact of the early experienced environment on individual animal behavior, and to compare it to that of the individuals original genetic predisposition. Additionally, we explored the correlation between personality traits measured indoors and the animals outdoor behavior. We studied Egyptian fruit bats, in which vast behavioral variability and plasticity have already been demonstrated. We raised bats in a captive colony under either enriched or impoverished environments and assessed their personality under controlled laboratory conditions. We then released the bats into the wild and tracked their foraging using GPS. Bats that had experienced an enriched environment during early life displayed increasing boldness and exploratory behavior when foraging outdoors, demonstrating how early-life experience can affect adult behavior. The individuals original predispositions did not predict their later foraging behavior. Our findings shed new light on the interplay between innate and experienced-based effects on individual behavior. Significance statementAnimals exhibit individual personalities, but how they are shaped is unknown. By employing controlled manipulations on new-born bat pups and GPS-tracking them outdoors as adults, we reveal that the early life conditions bats are exposed to have a significant impact on their foraging behavior as adults. We moreover show that the original individual predispositions of the bats do not predict their behavior. These findings enhance our understanding of the developmental factors that shape animal behavior and emphasize the vital importance of environmental enrichment during early life stages.

Group-living animals often coordinate their behavior using "contact calls". Identifying the function of these calls requires testing whether they are intended for any group member or targeted to specific preferred associates. If contact calling is used to coordinate with preferred associates, then higher rates of contact calling are expected between group members with a history of more frequent affiliation and cooperation. To test this hypothesis, we constructed a contact-calling network using synchronized recordings of vocal interactions between all 28 possible pairs of 8 female common vampire bats with well-sampled histories of social grooming and regurgitated food sharing. Bayesian multilevel models show that pairwise rates of contact calling were clearly predicted by social grooming and cooperative allofeeding rates in ways not explained by kinship. These findings show that common vampire bats use contact calls to coordinate with specific same-sex associates, unlike other studied bat species where individuals produce contact calls at similar rates towards different group members. We also found that, compared to white-winged vampire bats, common vampire bats are ten times less likely to rapidly respond to a contact call; this suggests yet-to-be-discovered differences in social behavior between vampire bat species. Finally, we discuss implications for the vocal grooming hypothesis.

For a century [1,2], studies of monozygotic and dizygotic twins have yielded estimates of trait heritability. The clever logic behind them is that while both types of twins share environments, their genetic overlap is different. Hence, larger trait correlations between monozygotic compared to dizygotic twins indicate heritability (nature), whereas similar correlations indicate shared environmental influences (nurture), and low correlations indicate shaping through non-shared environments (external influences and measurement error). While many have written on the assumptions that both types of twins share equal environments [3-5], and that parental genetics and environment are independent [6,7]; fewer have put their data where their mouth is. Here, the impacts of unmet assumptions were investigated using a generative mixture model of twin phenotypes. The results indicated that violations of the equal environments assumption yielded large overestimations of heritability and underestimations of shared environmental influences. On the other hand, when parental genetics shaped twins shared environments, only minor non-linear biases against heritability emerged. Finally, realistic levels of measurement error uniformly depressed estimates for genetic and shared environmental factors. In sum, twin studies are particularly susceptible to overestimation of genetic and non-shared environmental influences. This bias could explain why some traits, such as attitudes towards property taxes [8], show suspiciously high heritability without a biologically plausible mechanism. Particularly in the context of traits with convincing mechanisms of cultural transmission [9-11] and complex gene-environment interactions [6], researchers should not allow biases in twin studies to overestimate heritability.

The honey bee microbiome includes a wide variety of viruses. While most of them usually remain commensal, some can become pathogenic in specific contexts. Of these, one is that of deformed wing virus (DWV) and another, sacbrood virus (SBV). Although co-infection is the norm rather than the exception, most of the time these viruses have been studied independently. When investigated as co-infections, past studies have focused on their effects on the honey bee brood. In this study, we co-inoculated adult honey bees at emergence with DWV by injection and SBV orally (acting as the viral transmission by Varroa destructor and by trophallaxis or food, respectively), either simultaneously or sequentially. Using optical counters, we were able to track the survival and behaviour of these honey bees within colonies. Through regular in-hive sampling, we monitored the evolution of their viral loads as well as the expression of eight immune genes involved in honey bee anti-viral immunity. Here, we show that co-inoculations of DWV and SBV synergistically increase the virulence of DWV and conditionally promote the replication of both viruses. Finally, our results show that immune responses in adult honey bees depend on DWV genotypes and whether replication originates from a superinfecting virus or a virus already present in bees. Author SummaryHoney bees are highly social pollinators that live in crowded colonies. Their population density and the high frequency of interactions between individuals favours disease transmission and makes colonies susceptible to pathogen outbreaks. Many viruses commonly infect honey bees, however, they are often studied as single infections. As an effort to better understand interactions between honey bees and multiple viral populations, we co-inoculated young bees with two common honey bee viruses (deformed wing virus and sacbrood virus), released them in colonies and monitored their health and behaviour. Our findings show evidence of synergies between both viruses, as we show that a virus seemingly harmless for adult bees (sacbrood virus) may actually increase the virulence of another virus (deformed wing virus). These results highlight the importance of monitoring and studying multiple pathogens at once for a better understanding of the threat they represent to colony health and survival.

Inter- and intraspecific competition for resources is common among individuals which share ecological niches. To avoid physical confrontations, individuals can use various types of signals to demonstrate their dominance, including vocalizations. Kerivoula hardwickii is a solitary bat species that lives in highly ephemeral plant structures, which are therefore a limited resource. So far, it is unknown if individuals of K. hardwickii use vocalizations during competitive encounters for roosts, and if the intention of these vocalizations can be deduced by potential rivals. We hypothesized that the calls emitted during roost competition contain information that influences the ability of an individual to defend its roost. We conducted roost competition experiments in a flight cage, where there was an individual roost owner and an intruder who would attempt to evict the owner from the roost. All the vocalizations emitted during these encounters were recorded and analyzed to determine which acoustic parameters, if any, had an influence on the successful defense of the roost. We found that the calls emitted by males can influence their ability to defend the roost, and that entropy is the parameter that most strongly explains a successful defense. High entropy suggests that encounters between individuals of K. hardwickii escalate to high levels of aggressiveness and explain whether calls influence an individuals capacity to defend a roost. We suggest that bat vocalizations contain important information about individual characteristics, which in turn help bats make decisions during resource competition.

Bats are often the only mammals naturally colonizing isolated islands and are thus an excellent model to study evolutionary processes of insular ecosystems. Here, we studied the Reunion free-tailed bat (Mormopterus francoismoutoui), an endemic species to Reunion Island that has adapted to urban settings. At regional scale, we investigated the evolutionary history of Mormopterus species, as well as on Reunion Island sex-specific and seasonal patterns of genetic structure. We used an extensive spatio-temporal sampling including 1,136 individuals from 18 roosts and three biological seasons (non-reproductive/winter, pregnancy/summer, and mating), with additional samples from Mormopterus species from neighbouring islands (M. jugularis of Madagascar and M. acetabulosus of Mauritius). Complementary information gathered from both microsatellite and mitochondrial markers revealed a high genetic diversity but no signal of spatial genetic structure and weak evidence of female philopatry. Regional analysis suggests a single colonization event for M. francoismoutoui, dated around 175,000 years ago, and followed by in-situ diversification and the evolution of divergent ancestral lineages, which today form a large metapopulation. Population expansion was relatively ancient (55,000 years ago) and thus not linked to human colonization of the island and the availability of new anthropic day-roost sites. Discordant structure between mitochondrial and microsatellite markers suggests the presence of yet-unknown mating sites, or the recent evolution of putative ecological adaptations. Our study illustrates how understanding mechanisms involved in speciation can be challenging and the importance of both mitochondrial and nuclear DNA in resolving the wide in-situ diversification of an urban-dwelling bat, endemic to a small island.

The arctic archipelago of Svalbard (74 to 81{degrees} North) experiences extended periods of uninterrupted daylight in summer and uninterrupted darkness in winter. Species native to Svalbard display no daily rhythms in behaviour or physiology during these seasons, leading to the view that circadian rhythms may be redundant in arctic environments [1, 2]. Nevertheless, seasonal changes in the physiology and behaviour of arctic species rely on photoperiodic synchronisation to the solar year. Since this phenomenon is generally circadian-based in temperate species, we investigated if this might be a preserved aspect of arctic temporal organisation. Here, we demonstrate the involvement of the circadian clock in the seasonal photoperiodic response of the Svalbard ptarmigan (Lagopus muta hyperborea), the worlds northernmost resident bird species. First, we show the persistence of rhythmic clock gene expression under constant conditions within the mediobasal hypothalamus and pars tuberalis, the key tissues in the seasonal neuroendocrine cascade. We then employ a "sliding skeleton photoperiod" protocol, revealing that the driving force behind seasonal biology of the Svalbard ptarmigan is rhythmic sensitivity to light, a feature that depends on a functioning circadian rhythm. Our results suggest that the unusual selective pressure of the Arctic relaxes the adaptive value of the circadian clock for organisation of daily activity patterns, whilst preserving its importance for seasonal synchronisation. Thus, our data simultaneously reconnects circadian rhythms to life in the Arctic and establishes a universal principle of evolutionary value for circadian rhythms in seasonal biology.

Honey bees routinely fly hundreds or thousands of meters between their hive and established foraging locations [1-3]. To navigate these long distances, they are known to combine both landmark use [4-7] and path integration [8-11] and have been hypothesized to build a cognitive map [12-14]. Due to technical challenges inherent in tracking these small insects, obtaining three-dimensional, high-resolution measurements of individual navigational precision, and thus a detailed understanding of their strategies, has been difficult. Here, we utilize a novel multicopter drone-based tracking system [15] to measure the individual flight paths of honey bees in a structured agricultural landscape at unprecedented spatial and temporal resolution. Although bees could choose from multiple routes, we discovered that individual bees follow idiosyncratic paths with striking and repeatable precision. Flight path variability was highest over visually sparse regions and lowest near prominent proximal landmarks. Furthermore, individual strategies differed: some bees flew directly toward the hive before maneuvering around a specific tree, while others flew directly to a gap between a hedgerow and the tree. Thus, each animal varies in how it uses visual information and selects between behavioral strategies. The level of precision exhibited by their flight paths exceeds that reported in the waggle dance, implying that dance variability does not reflect a limit in the bees underlying spatial representation. Our results demonstrate the remarkable precision of individual bee navigation and illustrate the potential of this new drone-based tracking method to illuminate fine-scale behavioral mechanisms across a spectrum of honey bee ecology and cognition. HighlightsO_LIIndividual honey bees follow strikingly precise, idiosyncratic flight paths, revealing personalized strategies for navigating complex, obstacle-rich landscapes. C_LIO_LIVariability in individual bee flight paths was lower near conspicuous landmarks, but greater when further from such features, as a consequence variability in bee flights was not uniform along a route. C_LIO_LINavigational precision far exceeds waggle dance variability, showing that dance imprecision is not constrained by limits in spatial representation. C_LIO_LIMulticopter drone-based tracking method enables new investigations of 3D flight control and visual navigation in structurally complex realistic environments. C_LI