Predator Odor Research Articles

Traumatic stress-enhanced alcohol drinking: Sex differences and animal model perspectives.

Stress is associated with alcohol drinking, and epidemiological studies document the comorbidity of alcohol use disorder (AUD) and post-traumatic stress disorder (PTSD), with higher comorbid prevalence in females than in males. The aim of this paper is to highlight information related to sex differences in stress-enhanced alcohol drinking from clinical studies and from preclinical studies utilizing an animal model of traumatic stress. Stress is associated with alcohol drinking and relapse in males and females, but there are sex differences in the alcohol-related adaptation of stress pathways and in the association of different prefrontal regions with stress-induced anxiety. The predator stress model of traumatic stress produced enhanced alcohol drinking in a subgroup of stress-sensitive male and female animals, which could be associated with sex and subgroup differences in stress axis responsivity, behavioral responses to predator odors, and epigenetic mechanisms engaged by traumatic experiences. While additional studies in females are necessary, existing clinical and preclinical evidence suggests that biological mechanisms underlying stress-enhanced drinking likely differ between males and females. Thus, effective treatment strategies may differ between the sexes.

Differential behavioral response to predator odor in neuropathic pain in mice.

Neuropathic pain, a type of chronic pain caused by injury or disease of the somatosensory system, affects ∼10% of the general population and is difficult to treat. It is strongly associated with mood disorder comorbidities and impairs quality of life. It was recently suggested that hypervigilance caused by chronic pain might be of advantage in some species, helping them avoid predators during injury when they are most vulnerable. Here, we sought to confirm the hypervigilance hypothesis by using two predator odor (PO) paradigms, one with transient and one with continuous odor presentation. We observed behavioral responses to PO in neuropathic and control mice in an open field setting. We find that neuropathic mice show hypervigilance to PO, confirming previous results. However, we also find increased anxiety responses to neutral odor in neuropathic mice, which manifests as maladaptive pain. This demonstrates that this maladaptive nature of pain could be an evolutionary adaptation aimed at reducing injury-induced vulnerability.

No evidence of predator odor avoidance in a North American bird community

Recent advances in our understanding of avian chemical communication have highlighted the importance of olfaction in many aspects of avian life. Prior studies investigating predator avoidance behaviors in response to predator odor cues have produced mixed results across species and contexts. Here we assess if a community of birds in Eastern Pennsylvania displays avoidance behaviors towards predator odor cues in a natural foraging setting. We use clay caterpillars to measure foraging activity by birds in the presence of predator (bobcat) urine, non-predator (rabbit) urine, and water controls in two different environmental contexts (field vs. forest). Although we detected a weak trend for birds to forage less at predator urine-treated sites, we found no significant difference in avian foraging between the site types. We did find that foraging rates between environmental contexts changed significantly over the course of the experiment, with forest sites showing decreasing foraging rates and field sites showing increasing foraging rates. Our results reinforce the published literature that avoidance of predator odors by birds may not be ubiquitous across contexts and species.

Predator Odor Stressor, 2,3,5-Trimethyl-3-Thiazoline (TMT): Assessment of Stress Reactive Behaviors During an Animal Model of Traumatic Stress in Rats.

Animal models utilizing predator odor stress are important in understanding implications for post-traumatic stress disorder. 2,5-dihydro-2,4,5-trimethylthiazoline (TMT) has been used to measure stress reactive behaviors during TMT exposure, indicative of stress coping behaviors. In addition, long-term consequences of stress including contextual-induced stress memory, anxiety-like and hyperarousal behaviors, and subsequent increases in alcohol self-administration can also be examined after TMT exposure. In this article, we describe the TMT exposure protocol used in our lab and how we measure different stress-reactive behaviors that rats engage in during the TMT exposure. Rats are placed in Plexiglass chambers that contain white bedding on the bottom of the chamber and a metal basket in the top right corner containing a filter paper that 10 µl of TMT is pipetted onto. During the 10 min exposure, rats can move around the chamber freely. Exposures are recorded by a video camera for later analysis. During TMT exposure, rats engage in a variety of stress-reactive behaviors, including digging and immobility behavior. These are two distinctly different types of stress-induced behavioral coping strategies to measure individual differences in stress responsivity. To examine individual differences, we group rats into TMT-subgroups based on time spent engaging in digging or immobility behavior. We calculate a digging/immobility ratio score in which we divide the total time spent digging by the total time spent immobile. A cut-off strategy is used such that rats with a criterion ratio score <1.0 are classified as TMT-1 (i.e., low digging/high immobility; greater passive coping) and rats with a ratio score >1.0 are classified as TMT-2 (i.e., high digging/low immobility; greater active coping). Here, we provide a detailed description of the TMT exposure protocol and step-by-step process in evaluation of stress-reactive behaviors. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Predator odor stressor exposure using TMT Basic Protocol 2: Description of stress-reactive behaviors during TMT exposure and formation of TMT-subgroups.

Temperature during embryonic development in brown trout influences juvenile behaviour in encounters with predators

AbstractVariation in thermal conditions during embryogenesis can have far‐reaching impact throughout ontogeny and may give rise to behavioural variation. Many animals, such as salmonids, exhibit behavioural trade‐offs related to foraging and predator avoidance. How embryonic temperature affects these behaviours has remained unexplored. Not only abiotic conditions during embryogenesis but also biotic factors such as predator conditioning may affect fish behaviour, especially anti‐predator responses. We examined how elevated temperatures and predator odours throughout embryogenesis affect the behaviour of 28–37 mm young‐of‐the‐year brown trout (Salmo trutta) in encounters with predators, namely Atlantic salmon (Salmo salar; 20 cm) and burbot (Lota lota; 40 cm). Juvenile brown trout were more active and aggressive if they were incubated in warmer water as eggs than if they were incubated in colder water, and trout remained inactive longer when encountering predators if they were cold incubated. Brown trout were less active and aggressive when an Atlantic salmon was present than when a burbot or no predator was present. Behavioural responses did not differ between trout that had been subjected to water with versus without predator odours during embryogenesis, possibly because brown trout were not subjected to conspecific alarm cues during egg incubation. This study shows that thermal conditions during embryogenesis can influence fish behaviour early in life and thus contribute to behavioural variation, with potential effects on life history. Considering the rapid warming of northern regions, elevated embryonic temperatures may contribute substantially to variation in salmonid behaviour in the near future.

Exposure to the insecticide, imidacloprid, impairs predator-recognition learning in damselfly larvae

Many aquatic organisms use chemosensory information to learn about local predation threats, but contaminants in their environment may impair such cognitive processes. Neonicotinoids are a class of water-soluble systemic insecticides that have become a major concern in aquatic systems. In this study, we explored how a 10-day exposure to various concentrations (0, 0.1, 1.0, or 10.0 μg/L) of the neonicotinoid imidacloprid affects the learned recognition of predator odour by non-target damselfly larvae (Lestes spp). Unexposed larvae and those exposed to the low concentration (0.1 μg/L) demonstrated an appropriate learned response to a novel predator odour following a conditioning with the odour paired with chemical alarm cues. However, such learning failed to occur for larvae that were exposed to imidacloprid concentrations of 1.0 and 10.0 μg/L. Thus, either the cognitive processing of the chemical information was impaired or the chemistry of one or both of the conditioning cues was altered, making them ineffective for learning. In a second experiment, we found evidence for this latter hypothesis. In the absence of background imidacloprid exposure, larvae did not show significant learned responses to the predator odour when the conditioning cues were mixed with imidacloprid (initial pulse solution of 3.0 μg/L) at the start of conditioning (reaching a final concentration of 0.01 μg/L). These findings indicate that even low levels of imidacloprid can have important implications for chemosensory cognition of non-target species in aquatic environments.

Predator recognition learning by northern redbelly dace Chrosomus eos from a small kettle lake with a dynamic predator community

Predation, and the risk of predation, shape the ecology, behavior, and evolution of many species. Small fishes navigate a perilous landscape of risk in the shallow waters of the littoral zone. Moreover, in small lakes, the predator community can be dynamic due to stochastic colonization and extirpation events. These conditions select for the ability by small fish to acquire recognition of novel predators through associative learning. Chemical cues associated with predation events, such as damage-released chemical cues from conspecifics, and the odor of predators, inform prey of the presence of risk, and facilitate acquired recognition of novel predator odor. Deming Lake, MN, is a small meromictic lake with intermittent connections to neighboring lakes in the watershed. Annual sampling of the littoral fish community between the years 2000 and 2023 reveals a history of colonization and extirpation by relatively large-bodied species such as yellow perch Perca flavescens and pumpkinseed sunfish Lepomis gibbosus. Ice cover data, combined with limnological depth profiles of dissolved oxygen, confirm that dissolved oxygen is limiting during the winter and consistent with the hypothesis that extirpation of large-bodied species in Deming Lake is likely due to winter anoxia. These data set the stage for an experimental demonstration of acquired recognition of the odor of allopatric rock bass Ambloplites rupestris by bass-naïve northern redbelly dace Chrosomus eos from Deming Lake. Rock bass have been absent from Deming Lake since at least the year 2000. Predator-recognition learning allows redbelly dace, and many other small-bodied fishes that face variable predator species over ontogenetic, spatial, and temporal scales, a mechanism to adapt quickly to indicators of predation risk.

A nigroincertal projection mediates aversion and enhances coping responses to potential threat.

Recent studies have shown that the non-DA neurons in the ventral tegmental area (VTA) and substantia nigra (SN) not only modulate motivational behaviors but also regulate defensive behaviors. While zona incerta (ZI) is a threat-responsive substrate and receives innervations from the ventral midbrain, the function of the ventral midbrain-to-ZI connection remains poorly defined. Here, we demonstrate that the ZI receives heterogenous innervations from the ventral midbrain. By utilizing a retrograde AAV preferentially labeling non-DA neurons in the ventral midbrain, we found that ZI-projecting non-DA cells in the ventral midbrain are activated by restraint stress. We focused on the SN and found that SN-to-ZI GABAergic input is engaged by a predatory odor. Sustained pan-neuronal SN-to-ZI activation results in aversion and enhances defensive behaviors, likely through a disinhibition mechanism to recruit downstream brain regions that regulate defensive behaviors. Collectively, our results reveal a novel role of nigroincertal projection in mediating negative valence and regulating defensive behaviors.

Infant pain vs. pain with parental suppression: Immediate and enduring impact on brain, pain and affect.

In the short term, parental presence while a human infant is in pain buffers the immediate pain responses, although emerging evidence suggests repeated social buffering of pain may have untoward long-term effects. To explore the short- and long-term impacts of social buffering of pain, we first measured the infant rat pup's [postnatal day (PN) 8, or 12] response to mild tail shock with the mother present compared to shock alone or no shock. Shock with the mother reduced pain-related behavioral activation and USVs of pups at both ages and reduced Fos expression in the periaqueductal gray, hypothalamic paraventricular nucleus, and the amygdala at PN12 only. At PN12, shock with the mother compared to shock alone differentially regulated expression of several hundred genes related to G-protein-coupled receptors (GPCRs) and neural development, whereas PN8 pups showed a less robust and less coherent expression pattern. In a second set of experiments, pups were exposed to daily repeated Shock-mother pairings (or controls) at PN5-9 or PN10-14 (during and after pain sensitive period, respectively) and long-term outcome assessed in adults. Shock+mother pairing at PN5-9 reduced adult carrageenan-induced thermal hyperalgesia and reduced Fos expression, but PN10-14 pairings had minimal impact. The effect of infant treatment on adult affective behavior showed a complex treatment by age dependent effect. Adult social behavior was decreased following Shock+mother pairings at both PN5-9 and PN10-14, whereas shock alone had no effect. Adult fear responses to a predator odor were decreased only by PN10-14 treatment and the infant Shock alone and Shock+mother did not differ. Overall, integrating these results into our understanding of long-term programming by repeated infant pain experiences, the data suggest that pain experienced within a social context impacts infant neurobehavioral responses and initiates an altered developmental trajectory of pain and affect processing that diverges from experiencing pain alone.

Enhancement of peripheral fatty acyl ethanolamide signaling prevents stress-induced social avoidance and anxiety-like behaviors in male rats

Abstract Rationale Exposure to traumatic events can lead to alterations in social and anxiety-related behaviors. Emerging evidence suggests that peripheral host-defense processes are implicated in the expression of stress-induced behavioral responses and may be targeted to mitigate the negative sequalae of stress exposure. Objectives In this study, we used the peripherally restricted FAAH inhibitor URB937 to investigate the effects of the fatty acyl ethanolamide (FAE) family of lipid mediators – which include the endocannabinoid anandamide and the endogenous PPAR-α agonists, oleoylethanolamide and palmitoylethanolamide – on behavioral and peripheral biochemical responses to two ethologically distinct rat models of stress. Methods Male adult rats were exposed to acute social defeat, a model of psychological stress (Experiment 1), or to the predator odor 2,5-dihydro-2,4,5-trimethylthiazoline (TMT), a test of innate predator-evoked fear (Experiment 2), and subsequently treated with URB937 (1 or 3 mg/kg, intraperitoneal) or vehicle. Behavioral analyses were conducted 24 h (Experiment 1) or 7 days (Experiment 2) after exposure. Results URB937 administration prevented the emergence of both social avoidance behavior after social defeat stress and anxiety-related behaviors after TMT exposure. Further, URB937 administration blocked social defeat-induced transient increase in plasma concentrations of pro-inflammatory cytokines and the elevation in plasma corticosterone levels observed 24 h after social defeat Conclusions Enhancement of peripheral FAAH-regulated lipid signaling prevents the emergence of stress-induced social avoidance and anxiety-like behaviors in male rats through mechanisms that may involve an attenuation of peripheral cytokine release induced by stress exposure.

Traumatic Stress-Induced Increases in Anxiety-like Behavior and Alcohol Self-Administration Are Mediated by Central Amygdala CRF1 Neurons That Project to the Lateral Hypothalamus.

Avoidance stress coping, defined as persistent internal and/or external avoidance of stress-related stimuli, is a key feature of anxiety- and stress-related disorders, and contributes to increases in alcohol misuse after stress exposure. Previous work using a rat model of predator odor stress avoidance identified corticotropin-releasing factor (CRF) signaling via CRF Type 1 receptors (CRF1) in the CeA, as well as CeA projections to the lateral hypothalamus (LH) as key mediators of conditioned avoidance of stress-paired contexts and/or increased alcohol drinking after stress. Here, we report that CRF1-expressing CeA cells that project to the LH are preferentially activated in male and female rats that show persistent avoidance of predator odor stress-paired contexts (termed Avoider rats), and that chemogenetic inhibition of these cells rescues stress-induced increases in anxiety-like behavior and alcohol self-administration in male and female Avoider rats. Using slice electrophysiology, we found that prior predator odor stress exposure blunts inhibitory synaptic transmission and increases synaptic drive in CRF1 CeA-LH cells. In addition, we found that CRF bath application reduces synaptic drive in CRF1 CeA-LH cells in Non-Avoiders only. Collectively, these data show that CRF1 CeA-LH cells contribute to stress-induced increases in anxiety-like behavior and alcohol self-administration in male and female Avoider rats.SIGNIFICANCE STATEMENT Stress may lead to a variety of behavioral and physiological negative consequences, and better understanding of the neurobiological mechanisms that contribute to negative stress effects may lead to improved prevention and treatment strategies. This study, performed in laboratory rats, shows that animals that exhibit avoidance stress coping go on to develop heightened anxiety-like behavior and alcohol self-administration, and that these behaviors can be rescued by inhibiting the activity of a specific population of neurons in the central amygdala. This study also describes stress-induced physiological changes in these neurons that may contribute to their role in promoting increased anxiety and alcohol self-administration.

Predator odor can reduce acorn removal by granivorous rodents in mixed oak forest stands

Developing better practices for rodent pest control is of high importance to reduce damage during forest restoration and in crop production. For example, during direct seeding with large and highly attractive seeds such as acorns, most seeds will disappear due to consumption or dispersal if not protected. An unexplored concept in reducing rodent damage is the use of repellents derived from predators. We tested the efficiency of three volatile compounds (2-propylthietane, 2-phenylethylamine and indole) associated with predators as rodent repellent candidates and scored the reduction of acorn (Quercus petraea) removal at two field sites in southern Sweden. We further investigated at what distance (5 cm, 10 cm, and 15 cm) from the odor source the odors were efficient in lowering the removal of acorns. Removal was lowest with 2-propylthietane (25–45 % of acorn removal), followed by 2-phenylethylamine (75–95% acorn removal) at 5 cm. Indole failed to decrease acorn removal and did not differ significantly from the control treatments. In the control treatments, almost all acorns (95–100%) were removed from the plots during the 48-hour sessions at both sites, and the acorns were removed faster than in the other treatments. Removal increased with distance from the 2-propylthietane odor dispenser at both field sites. Here, the lowest acorn removal occurred at 5 cm from the odor disperser. Our short time experiment shows for the first time the potential of using 2-propylthietane for short-term protection of acorns from foraging by granivorous rodents in oak woodlands. To help increase the reliability of direct seeding as a method for regenerating and restoring forests, future studies should investigate whether the range and longevity in the field of predator odor formulations can be improved.

Do predator odours and warmer winters affect growth of salmonid embryos?

AbstractConditions early in ontogeny can have considerable effects later on in life. Many salmonids spawn during the autumn, and temperature during subsequent embryogenesis may have far‐reaching effects on life‐history traits, especially when considering ongoing climate change. Even biotic conditions during embryogenesis, such as predation threat, may affect later life stages. Here, we examined how predator odours and increased temperatures affect embryonic growth and development of a fish (brown trout Salmo trutta). We found that embryos had lower body mass and greater yolk volume close to hatching when subjected to predator odours. Trout embryos incubated at temperatures representing natural winter conditions were larger than embryos incubated at higher temperatures, although the latter hatched earlier. Fry sizes at emergence did not differ between treatments, perhaps because of compensatory growth during spring. This study shows that predator presence can have similar effects on embryonic growth of salmonids as warming winters, with possible impact later in ontogeny.

A sex-specific thermogenic neurocircuit induced by predator smell recruiting cholecystokinin neurons in the dorsomedial hypothalamus

Olfactory cues are vital for prey animals like rodents to perceive and evade predators. Stress-induced hyperthermia, via brown adipose tissue (BAT) thermogenesis, boosts physical performance and facilitates escape. However, many aspects of this response, including thermogenic control and sex-specific effects, remain enigmatic. Our study unveils that the predator odor trimethylthiazoline (TMT) elicits BAT thermogenesis, suppresses feeding, and drives glucocorticoid release in female mice. Chemogenetic stimulation of olfactory bulb (OB) mitral cells recapitulates the thermogenic output of this response and associated stress hormone corticosterone release in female mice. Neuronal projections from OB to medial amygdala (MeA) and dorsomedial hypothalamus (DMH) exhibit female-specific cFos activity toward odors. Cell sorting and single-cell RNA-sequencing of DMH identify cholecystokinin (CCK)-expressing neurons as recipients of predator odor cues. Chemogenetic manipulation and neuronal silencing of DMHCCK neurons further implicate these neurons in the propagation of predator odor-associated thermogenesis and food intake suppression, highlighting their role in female stress-induced hyperthermia.

Recruitment of Corticotropin-Releasing Hormone (CRH) Neurons in Categorically Distinct Stress Reactions in the Mouse Brain.

Corticotropin-releasing hormone (CRH) neurons in the paraventricular hypothalamic nucleus (PVH) are in the position to integrate stress-related information and initiate adaptive neuroendocrine-, autonomic-, metabolic- and behavioral responses. In addition to hypophyseotropic cells, CRH is widely expressed in the CNS, however its involvement in the organization of the stress response is not fully understood. In these experiments, we took advantage of recently available Crh-IRES-Cre;Ai9 mouse line to study the recruitment of hypothalamic and extrahypothalamic CRH neurons in categorically distinct, acute stress reactions. A total of 95 brain regions in the adult male mouse brain have been identified as containing putative CRH neurons with significant expression of tdTomato marker gene. With comparison of CRH mRNA and tdTomato distribution, we found match and mismatch areas. Reporter mice were then exposed to restraint, ether, high salt, lipopolysaccharide and predator odor stress and neuronal activation was revealed by FOS immunocytochemistry. In addition to a core stress system, stressor-specific areas have been revealed to display activity marker FOS. Finally, activation of CRH neurons was detected by colocalization of FOS in tdTomato expressing cells. All stressors resulted in profound activation of CRH neurons in the hypothalamic paraventricular nucleus; however, a differential activation of pattern was observed in CRH neurons in extrahypothalamic regions. This comprehensive description of stress-related CRH neurons in the mouse brain provides a starting point for a systematic functional analysis of the brain stress system and its relation to stress-induced psychopathologies.

Wistar-Kyoto rats and chronically stressed Wistar rats present similar depression- and anxiety-like behaviors but different corticosterone and endocannabinoid system modulation

The interplay of social, psychological, and biological stresses can trigger mental health conditions such as major depressive disorder (MDD), adjustment disorder, and posttraumatic stress disorder (PTSD). The endocannabinoid system (ECS), comprising endocannabinoids and cannabinoid receptors, is the critical pathway that mediates responses to stress stimuli. This study aimed to investigate the ECS's impact on responding to chronic social instability stress (SIS). Wistar (WIS) rats and an endogenously depressed rat model, Wistar-Kyoto (WKY), were used to evaluate depression- and anxiety-like behavioral responses, cognitive function, hormone levels, and ECS function. The animals in the stress group (WIS-STS and WKY-STS) were exposed to TMT (predator odor) for 10 mins (two exposures in total: one in light cycle and one in dark cycle) and daily roommate changes (30 days in total), while the control group (CTL) rats were exposed to a sham odor stimulus (distilled water) and did not undergo roommate changes.The results in the open field test suggest that WKY rats had significantly lower locomotor activity than WIS rats. In contrast, WKY rats and chronically stressed WIS rats presented similar depression- and anxiety-like behaviors and impaired cognitive function in the elevated plus maze, forced swimming test, and novel objective recognition test. However, chronic SIS did not exacerbate these behavioral changes in WKY rats. ELISA and Western blot analysis indicated that chronic SIS did not induce further upregulation of endocannabinoids and CB1R downregulation in WKY rats compared to WIS rats. In addition, the Luminex assay revealed that WKY rats showed a higher resilience on the HPA-axis modulation towards chronic SIS, distinguished by the hyperactivity of the HPA-axis modulation in WIS rats.Overall, the study revealed that the chronic SIS animal model (stressed WIS rats) and an animal model of endogenous depression (WKY rats) can generate similar behavioral changes in anxious behavior, behavioral despair, and cognitive impairment. Both animal models present hyperactivity of the ACTH modulation and ECS activity, while WKY rats are more resilient on CORT modulation towards chronic SIS.

Global Neuropeptidome Profiling in Response to Predator Stress in Rat: Implications for Post-Traumatic Stress Disorder.

Traumatic stress triggers or exacerbates multiple psychiatric illnesses, including post-traumatic stress disorder (PTSD). Nevertheless, the neurophysiological mechanisms underlying stress-induced pathology remain unclear, in part due to the limited understanding of neuronal signaling molecules, such as neuropeptides, in this process. Here, we developed mass spectrometry (MS)-based qualitative and quantitative analytical strategies to profile neuropeptides in rats exposed to predator odor (an ethologically relevant analogue of trauma-like stress) versus control subjects (no odor) to determine peptidomic alterations induced by trauma. In total, 628 unique neuropeptides were identified across 5 fear-circuitry-related brain regions. Brain-region-specific changes of several neuropeptide families, including granin, ProSAAS, opioids, cholecystokinin, and tachykinin, were also observed in the stressed group. Neuropeptides from the same protein precursor were found to vary across different brain regions, indicating the site-specific effects of predator stress. This study reveals for the first time the interaction between neuropeptides and traumatic stress, providing insights into the molecular mechanisms of stress-induced psychopathology and suggesting putative novel therapeutic strategies for disorders such as PTSD.

Habitat complexity and predator odours impact on the stress response and antipredation behaviour in coral reef fish.

Mass coral bleaching events coupled with local stressors have caused regional-scale loss of corals on reefs globally. Following the loss of corals, the structural complexity of these habitats is often reduced. By providing shelter, obscuring visual information, or physically impeding predators, habitat complexity can influence predation risk and the perception of risk by prey. Yet little is known on how habitat complexity and risk assessment interact to influence predator-prey interactions. To better understand how prey's perception of threats may shift in degraded ecosystems, we reared juvenile Pomacentrus chrysurus in environments of various habitat complexity levels and then exposed them to olfactory risk odours before simulating a predator strike. We found that the fast-start escape responses were enhanced when forewarned with olfactory cues of a predator and in environments of increasing complexity. However, no interaction between complexity and olfactory cues was observed in escape responses. To ascertain if the mechanisms used to modify these escape responses were facilitated through hormonal pathways, we conducted whole-body cortisol analysis. Cortisol concentrations interacted with habitat complexity and risk odours, such that P. chrysurus exhibited elevated cortisol levels when forewarned with predator odours, but only when complexity levels were low. Our study suggests that as complexity is lost, prey may more appropriately assess predation risk, likely as a result of receiving additional visual information. Prey's ability to modify their responses depending on the environmental context suggests that they may be able to partly alleviate the risk of increased predator-prey interactions as structural complexity is reduced.

Predatory arthropods and cues associated with predator presence elicit strong non‐consumptive effects on bluegrass billbugs

AbstractTurfgrasses hosts a diverse community of predators that contribute to herbivore suppression. However, there is little evidence that predators directly consume a common turfgrass herbivore, the billbug (Sphenophorus sp.) (Coleoptera: Curculionidae). Given low rates of direct consumption by predators, prospects for strong non‐consumptive effects in billbugs were underappreciated. However, this assumed lack of non‐consumptive effects presupposes that herbivorous prey, like billbugs, can identify predators and the level of threat that they pose. Here, we examined this assumption and quantified the extent to which exposure to predators and predator cues altered the behavior (predator avoidance, feeding, and mating) of bluegrass billbug, Sphenophorus parvulus Gyllenhal. First, using a series of behavioral assays within microcosms, we exposed billbugs to predators – Pterostichus melanarius (Illiger) (Coleoptera: Carabidae), Harpalus sp. (Coleoptera: Carabidae), Philonthus sp. (Coleoptera: Staphylinidae), and Hogna sp. (Araneae: Lycosidae) – singly and in combination to measure the effect of contact with predators and potential synergy among predator species pairs. Indirect exposure to predators, predator odor, and non‐predatory arthropods were also used to measure effects of various predator cues on billbug behavior. In subsequent Y‐tube choice assays we determined whether billbugs discriminated between filtered air and predator odor to isolate chemical signaling as a potential mechanism for billbug predator detection. Direct exposure to predators increased billbug predator avoidance (up to 201%), and decreased billbug feeding (down to −78%), though no predation was observed. Similar behavioral changes were observed when billbugs were exposed indirectly to a predator, and to predator odor. In Y‐tube assays billbugs avoided predator odor, further suggesting billbugs use chemical cues to detect predator presence and that these may be a driver in their observed behavior. Our results show that billbugs exhibit strong non‐consumptive effects, even in the absence of strong direct consumptive effects. These results provide the foundation for further research concerning the role of chemical signaling in billbug predator detection.

Detection of neuronal defensive discharge information transmission and characteristics in periaqueductal gray double-subregions using PtNP/PEDOT:PSS modified microelectrode arrays

Threatened animals respond with appropriate defensive behaviors to survive. It has been accepted that midbrain periaqueductal gray (PAG) plays an essential role in the circuitry system and organizes defensive behavioral responses. However, the role and correlation of different PAG subregions in the expression of different defensive behaviors remain largely unexplored. Here, we designed and manufactured a microelectrode array (MEA) to simultaneously detect the activities of dPAG and vPAG neurons in freely behaving rats. To improve the detection performance of the MEAs, PtNP/PEDOT:PSS nanocomposites were modified onto the MEAs. Subsequently, the predator odor was used to induce the rat’s innate fear, and the changes and information transmission in neuronal activities were detected in the dPAG and vPAG. Our results showed that the dPAG and vPAG participated in innate fear, but the activation degree was distinct in different defense behaviors. During flight, neuronal responses were stronger and earlier in the dPAG than the vPAG, while vPAG neurons responded more strongly during freezing. By applying high-performance MEA, it was revealed that neural information spread from the activated dPAG to the weakly activated vPAG. Our research also revealed that dPAG and vPAG neurons exhibited different defensive discharge characteristics, and dPAG neurons participated in the regulation of defense responses with burst-firing patterns. The slow activation and continuous firing of vPAG neurons cooresponded with the regulation of long-term freezing responses. The results demonstrated the important role of PAG neuronal activities in controlling different aspects of defensive behaviors and provided novel insights for investigating defense from the electrophysiological perspective.