Defensive behavior: Sensing threats from terrestrial and aerial predators.

The dorsal periaqueductal gray (dPAG) contains a tonically GABAergic network controlling defensive responses. Determining how this intrinsic dPAG inhibitory circuit functions might provide critical insights into how anti-predatory responses are organized.

Urban birds' tolerance towards humans was largely unaffected by COVID-19 shutdown-induced variation in human presence.

The coronavirus disease 2019 (COVID-19) pandemic and respective shutdowns dramatically altered human activities, potentially changing human pressures on urban-dwelling animals. Here, we use such COVID-19-induced variation in human presence to evaluate, across multiple temporal scales, how urban birds from five countries changed their tolerance towards humans, measured as escape distance. We collected 6369 escape responses for 147 species and found that human numbers in parks at a given hour, day, week or year (before and during shutdowns) had a little effect on birds' escape distances. All effects centered around zero, except for the actual human numbers during escape trial (hourly scale) that correlated negatively, albeit weakly, with escape distance. The results were similar across countries and most species. Our results highlight the resilience of birds to changes in human numbers on multiple temporal scales, the complexities of linking animal fear responses to human behavior, and the challenge of quantifying both simultaneously in situ.

Tourism influences escape behavior of lizards in relationship with human clothing color.

Increased tourism pressure modifies animal behavior, including alterations in anti-predator responses and foraging activity. In areas with high tourist presence, animals may become accustomed to increased human activity and adjust the intensity of some defensive responses. An animal's anti-predation ability is usually estimated by measuring its Alert Initiation Distance (AID) and Flight Initiation Distance (FID). Both indexes are affected by multiple factors including the color of the observer's clothing. Animal behavior is also influenced by human presence, and individuals may become accustomed to increased human presence, e.g. in tourist areas. In this study, we analysed the escape behavior of the endemic Cyprus rock agama (Laudakia cypriaca) in relation to the observers clothing color. Our results showed that AIDs and FIDs of agamas in tourist areas were significantly shorter than those in non-tourist areas. Moreover, in non-tourist areas, AIDs and FIDs of agamas were significantly longer when the observer wore red clothes, compared to green and grey clothes. Our results may be helpful in planning research taking into account various colored clothing based on expected reptilian reactions. Furthermore, our results may determine the proximity at which humans interact with animals, considering clothing color, to prevent negative impacts especially on rare and protected lizard species.

The human hypothalamus coordinates switching between different survival actions.

Comparative research suggests that the hypothalamus is critical in switching between survival behaviors, yet it is unclear if this is the case in humans. Here, we investigate the role of the human hypothalamus in survival switching by introducing a paradigm where volunteers switch between hunting and escape in response to encounters with a virtual predator or prey. Given the small size and low tissue contrast of the hypothalamus, we used deep learning-based segmentation to identify the individual-specific hypothalamus and its subnuclei as well as an imaging sequence optimized for hypothalamic signal acquisition. Across 2 experiments, we employed computational models with identical structures to explain internal movement generation processes associated with hunting and escaping. Despite the shared structure, the models exhibited significantly different parameter values where escaping or hunting were accurately decodable just by computing the parameters of internal movement generation processes. In experiment 2, multi-voxel pattern analyses (MVPA) showed that the hypothalamus, hippocampus, and periaqueductal gray encode switching of survival behaviors while not encoding simple motor switching outside of the survival context. Furthermore, multi-voxel connectivity analyses revealed a network including the hypothalamus as encoding survival switching and how the hypothalamus is connected to other regions in this network. Finally, model-based fMRI analyses showed that a strong hypothalamic multi-voxel pattern of switching is predictive of optimal behavioral coordination after switching, especially when this signal was synchronized with the multi-voxel pattern of switching in the amygdala. Our study is the first to identify the role of the human hypothalamus in switching between survival behaviors and action organization after switching.

Tonically active GABAergic neurons in the dorsal periaqueductal gray control instinctive escape in mice.

Escape behavior is a set of locomotor actions that move an animal away from threat. While these actions can be stereotyped, it is advantageous for survival that they are flexible.1,2,3 For example, escape probability depends on predation risk and competing motivations,4,5,6,7,8,9,10,11 and flight to safety requires continuous adjustments of trajectory and must terminate at the appropriate place and time.12,13,14,15,16 This degree of flexibility suggests that modulatory components, like inhibitory networks, act on the neural circuits controlling instinctive escape.17,18,19,20,21,22 In mice, the decision to escape from imminent threats is implemented by a feedforward circuit in the midbrain, where excitatory vesicular glutamate transporter 2-positive (VGluT2+) neurons in the dorsal periaqueductal gray (dPAG) compute escape initiation and escape vigor.23,24,25 Here we tested the hypothesis that local GABAergic neurons within the dPAG control escape behavior by setting the excitability of the dPAG escape network. Using in vitro patch-clamp and in vivo neural activity recordings, we found that vesicular GABA transporter-positive (VGAT+) dPAG neurons fire action potentials tonically in the absence of synaptic inputs and are a major source of inhibition to VGluT2+ dPAG neurons. Activity in VGAT+ dPAG cells transiently decreases at escape onset and increases during escape, peaking at escape termination. Optogenetically increasing or decreasing VGAT+ dPAG activity changes the probability of escape when the stimulation is delivered at threat onset and the duration of escape when delivered after escape initiation. We conclude that the activity of tonically firing VGAT+ dPAG neurons sets a threshold for escape initiation and controls the execution of the flight action.

Nearest neighbour distance does not affect escape behaviour in urban hooded crows.

The nearest-neighbour distance is an important property of a group, as individuals can obtain environmental information more quickly and easily from nearby individuals. We examined whether distance to the nearest neighbour affected two components of escape behaviour - alert distance (AD) and flight initiation distance (FID) - in an urban population of hooded crows Corvus cornix, while controlling for confounding variables. We did not find evidence that AD and FID were influenced by the nearest neighbour distance. However, both AD and FID were negatively affected by feeding activity of individuals - focal crows alerted later and escaped at shorter distance if they were feeding during our approach. In addition, AD and FID were positively related to starting distance and grass coverage. The lack of evidence for the nearest neighbour effect on escape behaviour of crows may be due to: (1) disturbance by close neighbours that may impede antipredator behaviour of focal birds, (2) variable distribution of familiar, dominant or experienced individuals within a flock, and (3) dynamic change in position of the nearest neighbour during the potential predator approach.

Surprising multifunctionality of a Tritonia swim CPG neuron: C2 drives the early phase of postswim crawling despite being silent during the behavior.

In response to a suitably aversive skin stimulus, the marine mollusk Tritonia diomedea launches an escape swim followed by several minutes of high-speed crawling. The two escape behaviors are highly dissimilar: whereas the swim is a muscular behavior involving alternating ventral and dorsal whole body flexions, the crawl is a nonrhythmic gliding behavior mediated by the beating of foot cilia. The serotonergic dorsal swim interneurons (DSIs) are members of the swim central pattern generator (CPG) and also strongly drive crawling. Although the swim network is very well understood, the Tritonia crawling network to date comprises only three neurons: the DSIs and pedal neurons 5 and 21 (Pd5 and Pd21). Since Tritonia's swim network has been suggested to have arisen from a preexisting crawling network, we examined the possible role that another swim CPG neuron, C2, may play in crawling. Because of its complete silence in the postswim crawling period, C2 had not previously been considered to play a role in driving crawling. However, semi-intact preparation experiments demonstrated that a brief C2 spike train surprisingly and strongly drives the foot cilia for ∼30 s, something that cannot be explained by its synaptic connections to Pd5 and Pd21. Voltage-sensitive dye (VSD) imaging in the pedal ganglion identified many candidate crawling motor neurons that fire at an elevated rate after the swim and also revealed several pedal neurons that are strongly excited by C2. It is intriguing that unlike the DSIs, which fire tonically after the swim to drive crawling, C2 does so despite its postswim silence.NEW & NOTEWORTHY Tritonia swim central pattern generator (CPG) neuron C2 surprisingly and strongly drives the early phase of postswim crawling despite being silent during this period. In decades of research, C2 had not been suspected of driving crawling because of its complete silence after the swim. Voltage-sensitive dye imaging revealed that the Tritonia crawling motor network may be much larger than previously known and also revealed that many candidate crawling neurons are excited by C2.

Synergistic effects of musking and autohaemorrhaging on the duration of death feigning in dice snakes (Natrix tessellata).

Predation exerts a significant selection pressure on prey, shaping a multitude of traits that serve as antipredator defences. In turn, natural selection could favour combinations of antipredator defences with synergistic effects that enhance prey survival. An especially interesting antipredator defence is death feigning (DF), present in a wide variety of taxa and usually characterized by the prey lying motionless often along with defaecation, musking and autohaemorrhaging (AH). All these aspects of the DF display should work in conjunction with one another, intensifying the overall effect of the display and in turn facilitating quicker escape. To confirm this hypothesis, we tested 263 dice snakes (Natrix tessellata) directly in the field. We noted the occurrence of smearing faeces, musk and AH, and we measured the duration of DF, expecting to see a negative association between the occurrence of these behaviours and the duration of DF. Our results affirm our hypothesis: dice snakes that smeared themselves in musk and faeces prior to DF and had AH during DF spent significantly less time in DF. Our results highlight the functional integration of antipredator behaviours across different phases of predator-prey interactions, emphasizing the need for future research to prioritize studying the sequential display of behaviours.

A robotic falcon induces similar collective escape responses in different bird species.

Patterns of collective escape of a bird flock from a predator are fascinating, but difficult to study under natural conditions because neither prey nor predator is under experimental control. We resolved this problem by using an artificial predator (RobotFalcon) resembling a peregrine falcon in morphology and behaviour. We imitated hunts by chasing flocks of corvids, gulls, starlings and lapwings with the RobotFalcon, and compared their patterns of collective escape to those when chased by a conventional drone and, in case of starlings, hunted by wild peregrine falcons. Active pursuit of flocks, rather than only flying nearby by either the RobotFalcon or the drone, made flocks collectively escape more often. The RobotFalcon elicited patterns of collective escape in flocks of all species more often than the drone. Attack altitude did not affect the frequency of collective escape. Starlings escaped collectively equally often when chased by the RobotFalcon or a wild peregrine falcon. Flocks of all species reacted most often by collective turns, second most often by compacting and third by splitting into subflocks. This study demonstrates the potential of an artificial aerial predator for studying the collective escape behaviour of free-living birds, opening exciting avenues in the empirical study of prey-predator interactions.

Elevated ammonia cues hatching in red-eyed treefrogs: A mechanism for escape from drying eggs.

Egg dehydration can kill terrestrial frog embryos, and this threat is increasing with climate change and deforestation. In several lineages that independently evolved terrestrial eggs, and retained aquatic tadpoles, embryos accelerate hatching to escape from drying eggs, entering the water earlier and less developed. However, the cues that stimulate drying-induced early hatching are unknown. Ammonia is a toxic, water-soluble metabolic waste that accumulates within eggs as embryos develop and concentrates as eggs dehydrate. Thus, increasing ammonia concentration may be a direct threat to embryos in drying eggs. We hypothesized that it could serve as a cue, stimulating embryos to hatch and escape. The embryos of red-eyed treefrogs, Agalychnis callidryas, hatch early to escape from many threats, including dehydration, and are known to use mechanosensory, hypoxia, and light cues. To test if they also use high ammonia as a cue to hatch, we exposed stage-matched pairs of hatching-competent, well-hydrated sibling embryos to ammonia and control solutions in shallow water baths and recorded their behavior. Control embryos remained unhatched while ammonia-exposed embryos showed a rapid, strong hatching response; 95% hatched, on average in under 15 min. This demonstrates that elevated ammonia can serve as a hatching cue for A. callidryas embryos. This finding is a key step in understanding the mechanisms that enable terrestrial frog embryos to escape from egg drying, opening new possibilities for integrative and comparative studies on this growing threat.

Automated escape system: identifying prey's kinematic and behavioral features critical for predator evasion.

Identifying the kinematic and behavioral variables of prey that influence evasion from predator attacks remains challenging. To address this challenge, we have developed an automated escape system that responds quickly to an approaching predator and pulls the prey away from the predator rapidly, similar to real prey. Reaction distance, response latency, escape speed and other variables can be adjusted in the system. By repeatedly measuring the response latency and escape speed of the system, we demonstrated the system's ability to exhibit fast and rapid responses while maintaining consistency across successive trials. Using the live predatory fish species Coreoperca kawamebari, we show that escape speed and reaction distance significantly affect the outcome of predator-prey interactions. These findings indicate that the developed escape system is useful for identifying kinematic and behavioral features of prey that are critical for predator evasion, as well as for measuring the performance of predators.

Single administration of a psychedelic [(R)-DOI] influences coping strategies to an escapable social stress.

Psychedelic compounds have potentially rapid, long-lasting anxiolytic, antidepressive and anti-inflammatory effects. We investigated whether the psychedelic compound (R)-2,5-dimethoxy-4-iodoamphetamine [(R)-DOI], a selective 5-HT2A receptor partial agonist, decreases stress-related behavior in male mice exposed to repeated social aggression. Additionally, we explored the likelihood that these behavioral changes are related to anti-inflammatory properties of [(R)-DOI]. Animals were subjected to the Stress Alternatives Model (SAM), an escapable social stress paradigm in which animals develop reactive coping strategies - remaining in the SAM arena (Stay) with a social aggressor, or dynamically initiated stress coping strategies that involve utilizing the escape holes (Escape) to avoid aggression. Mice expressing these behavioral phenotypes display behaviors like those in other social aggression models that separate animals into stress-vulnerable (as for Stay) or stress-resilient (as for Escape) groups, which have been shown to have distinct inflammatory responses to social stress. These results show that Stay animals have heightened cytokine gene expression, and both Stay and Escape mice exhibit plasma and neural concentrations of the inflammatory cytokine tumor necrosis factor-α (TNFα) compared to unstressed control mice. Additionally, these results suggest that a single administration of (R)-DOI to Stay animals in low doses, can increase stress coping strategies such as increasing attention to the escape route, promoting escape behavior, and reducing freezing during socially aggressive interaction in the SAM. Lower single doses of (R)-DOI, in addition to shifting behavior to suggest anxiolytic effects, also concomitantly reduce plasma and limbic brain levels of the inflammatory cytokine TNFα.

Sealing the deal - Antarctic fur seals' active hunting tactics to capture small evasive prey revealed by miniature sonar tags.

The ability of predators to adopt hunting tactics that minimise escape reactions from prey is crucial for efficient foraging, and depends on detection capabilities and locomotor performance of both predators and prey. Here, we investigated the efficiency of a small pinniped, the Antarctic fur seal (Arctocephalus gazella) at exploiting their small prey by describing for the first time their fine-scale predator-prey interactions. We compared these with those from another diving predator, the southern elephant seal (Mirounga leonina) that forage on the same prey type. We used data recorded by a newly developed sonar tag that combines active acoustics with ultrahigh-resolution movement sensors to study simultaneously the fine-scale behaviour of both Antarctic fur seals and prey during predator-prey interactions in more than 1200 prey capture events for eight female Antarctic fur seals. Our results showed that Antarctic fur seals and their prey detect each other at the same time, i.e. 1-2 s before the strike, forcing Antarctic fur seals to display reactive fast-moving chases to capture their prey. In contrast, southern elephant seals detect their prey up to 10 s before the strike, allowing them to approach their prey stealthily without triggering an escape reaction. The active hunting tactics used by Antarctic fur seals is probably very energy consuming compared with the stalking tactics used by southern elephant seals but might be compensated for by the consumption of faster-moving larger prey. We suggest that differences in manoeuvrability, locomotor performance and detection capacities and in pace of life between Antarctic fur seals and southern elephant seals might explain these differences in hunting styles.

Recent advances in insect vision in a 3D world: looming stimuli and escape behaviour.

Detecting looming motion directly towards the insect is vital to its survival. Looming detection in two insects, flies and locusts, is described and contrasted. Pathways using looming detectors to trigger action and their topographical layout in the brain is explored in relation to facilitating behavioural selection. Similar visual stimuli, such as looming motion, are processed by nearby glomeruli in the brain. Insect-inspired looming motion detectors are combined to detect and avoid collision in different scenarios by robots, vehicles and unmanned aerial vehicle (UAV)s.

A method for studying escape behavior to terrestrial threats in rodents.

BACKGROUND: Escape is one of the most essential behaviors for an animal's survival because it could be a matter of life and death. Much of our current understanding of the neural mechanisms underlying escape is derived from the looming paradigm, which mimics a diving aerial predator. Yet, the idea of the looming paradigm does not account for all types of threats like lions hunting antelopes or cats stalking mice. Escape responses to such terrestrial threats may require different strategies and neural mechanisms. NEW METHODS: Here, we developed a real-time interactive platform to study escape behavior to terrestrial threats in mice. A closed-loop controlled robot was magnetically pulled to mimic a terrestrial threat that chases a mouse. By using strong magnets and high-precision servo motors, the robot is capable of moving precisely with a high spatial-temporal resolution. Different algorithms can be used to achieve single approach or persistent approach. RESULTS: Animal experiments showed that mice exhibited consistent escape behavior when exposed to an approaching robotic predator. When presented with a persistently approaching predator, the mice were able to rapidly adapt their behavior, as evidenced by a decrease in startle responses and changes in movement patterns. COMPARISON WITH EXISTING METHODS: In comparison to existing methods for studying escape behavior, such as the looming paradigm, this approach is more suitable for investigating animal behavior in response to sustained threats. CONCLUSION: In conclusion, we have developed a flexible platform to study escape behavior to terrestrial threats in mice.

Escape performance in the cyclopoid copepod Oithona davisae.

Escaping a predator is one of the keys to success for any living creature. The performance of adults (males, females, and ovigerous females) of the cyclopoid copepod Oithona davisae exposed to an electrical stimulus is analysed as a function of temperature by measuring characteristic parameters associated with the escape movement (distance covered, duration of the appendage movement, mean and maximum escape speeds, Reynolds number). In addition, as a proxy for the efficiency of the motion, the Strouhal number was calculated. The escape performance showed temperature-dependent relationships within each adult state, as well as differences between sexes; additionally, changes owing to the presence of the egg sac were recorded in females. In a broader perspective, the results collected reveal the occurrence of different behavioural adaptations in males and females, adding to the comprehension of the mechanisms by which O. davisae interacts with its environment and shedding new light on the in situ population dynamics of this species.

Neuromuscular basis of Drosophila larval rolling escape behavior.

When threatened by dangerous or harmful stimuli, animals engage in diverse forms of rapid escape behaviors. In Drosophila larvae, one type of escape response involves C-shaped bending and lateral rolling followed by rapid forward crawling. The sensory circuitry that promotes larval escape has been extensively characterized; however, the motor programs underlying rolling are unknown. Here, we characterize the neuromuscular basis of rolling escape behavior. We used high-speed, volumetric, Swept Confocally Aligned Planar Excitation (SCAPE) microscopy to image muscle activity during larval rolling. Unlike sequential peristaltic muscle contractions that progress from segment to segment during forward and backward crawling, muscle activity progresses circumferentially during bending and rolling escape behavior. We propose that progression of muscular contraction around the larva's circumference results in a transient misalignment between weight and the ground support forces, which generates a torque that induces stabilizing body rotation. Therefore, successive cycles of slight misalignment followed by reactive aligning rotation lead to continuous rolling motion. Supporting our biomechanical model, we found that disrupting the activity of muscle groups undergoing circumferential contraction progression leads to rolling defects. We use EM connectome data to identify premotor to motor connectivity patterns that could drive rolling behavior and perform neural silencing approaches to demonstrate the crucial role of a group of glutamatergic premotor neurons in rolling. Our data reveal body-wide muscle activity patterns and putative premotor circuit organization for execution of the rolling escape response.

Improving the success of reinforcement programs: effects of a two-week confinement in a field enclosure on the anti-predator behaviour of captive-bred European hamsters.

Captive breeding programs are an important pillar in biodiversity conservation, aiming to prevent the extinction of threatened species. However, the establishment of self-sustaining populations in the wild through the release of captive-bred animals is often hampered by a high mortality upon release. In this study, we investigated how a 2-week confinement period within a large field enclosure affected the anti-predator behaviour of 'naive' captive-bred hamsters and how potential modifications persisted over time. During three consecutive tests, hamsters were confronted with a moving predator model (a red fox mount, Vulpes vulpes) and their behaviour was filmed. After the initial round of confrontation with the predator model, one group of hamsters (field group) was released into a field enclosure protected from predators, while the other group (control) remained in their individual laboratory cages. After 2 weeks, hamsters from the field group were recaptured and individuals of both groups underwent a second confrontation test. A total of 1 month after their return from the field enclosure, field hamsters were subjected to a last confrontation test. Video analysis, investigating four behavioural variables, revealed that field hamsters significantly modified their behavioural response following the 2 weeks confinement in the enclosure, while this was not the case for control hamsters. In addition, most behavioural modifications in field hamsters persisted over 1 month, while others started to revert. We suggest that an appropriate pre-release period inside a field enclosure will enable naive (captive-bred) hamsters to develop an adequate anti-predator behaviour that will increase their immediate survival probability upon release into the wild. We believe that such measure will be of great importance for hamster conservation programs.

No evidence for magnetic field effects on the behaviour of Drosophila.

Migratory songbirds have the remarkable ability to extract directional information from the Earth's magnetic field1,2. The exact mechanism of this light-dependent magnetic compass sense, however, is not fully understood. The most promising hypothesis focuses on the quantum spin dynamics of transient radical pairs formed in cryptochrome proteins in the retina3-5. Frustratingly, much of the supporting evidence for this theory is circumstantial, largely because of the extreme challenges posed by genetic modification of wild birds. Drosophila has therefore been recruited as a model organism, and several influential reports of cryptochrome-mediated magnetic field effects on fly behaviour have been widely interpreted as support for a radical pair-based mechanism in birds6-23. Here we report the results of an extensive study testing magnetic field effects on 97,658 flies moving in a two-arm maze and on 10,960 flies performing the spontaneous escape behaviour known as negative geotaxis. Under meticulously controlled conditions and with vast sample sizes, we have been unable to find evidence for magnetically sensitive behaviour in Drosophila. Moreover, after reassessment of the statistical approaches and sample sizes used in the studies that we tried to replicate, we suggest that many-if not all-of the original results were false positives. Our findings therefore cast considerable doubt on the existence of magnetic sensing in Drosophila and thus strongly suggest that night-migratory songbirds remain the organism of choice for elucidating the mechanism of light-dependent magnetoreception.

Kinematics and behaviour in fish escape responses: guidelines for conducting, analysing and reporting experiments.

Work carried out since the late 1970s has provided key insights into the comparative biomechanics, kinematics, behaviour and neurobiology of fish escape responses. An escape response is an ecologically important behaviour used by fishes to evade predation and aggression via rapid swimming movements. With environmental change expected to affect the physiology and biomechanics of aquatic ectotherms, there is a growing interest in understanding how environmental stressors affect the swimming performance and behaviour of fishes during escape responses, particularly in the context of predator-prey interactions. As the study of fish swimming continues to expand, there have been repeated calls to standardise experiments and reporting practices to facilitate integrative and comparative studies. Here, we provide a set of practical guidelines for conducting, analysing and reporting experiments on escape responses in fish, including a reporting checklist to assist authors undertaking these experiments. These resources will facilitate executing and reporting escape response experiments in a rigorous and transparent fashion, helping to advance the study of fish swimming in an era of rapid environmental change.