Binding Affinity, Selectivity, and Pharmacokinetics of the Oxytocin Receptor Antagonist L-368,899 in the Coyote (Canis latrans).

L-368,899 is a selective small-molecule oxytocin receptor (OXTR) antagonist originally developed in the 1990s to prevent preterm labor. Although its utility for that purpose was limited, L-368,899 is now one of the most commonly used drugs in animal research for the selective blockade of neural OXTR after peripheral delivery. A growing number of rodent and primate studies have used L-368,899 to evaluate whether certain behaviors are oxytocin dependent. These studies have improved our understanding of oxytocin's function in the brains of rodents and monkeys, but very little work has been done in other mammals, and only a single paper in macaques has provided any evidence that L-368,899 can be detected in the CNS after peripheral delivery. The current study sought to extend those findings in a novel species: coyotes ( Canis latrans ). Coyotes are ubiquitous North American canids that form long-term monogamous pair-bonds. Although monogamy is rare in rodents and primates, all wild canid species studied to date exhibit social monogamy. Coyotes are therefore an excellent model organism for the study of oxytocin and social bonds. Our goal was to determine whether L-368,899 is a viable candidate for future use in behavioral studies in coyotes. We used captive coyotes at the USDA National Wildlife Research Center's Predator Research Facility to evaluate the pharmacokinetics of L-368,899 in blood and CSF during a 90-min time course after intramuscular injection. We then characterized the binding affinity and selectivity of L-368,899 to coyote OXTR and the structurally similar vasopressin 1a receptor. We found that L-368,899 peaked in CSF at 15 to 30 min after intramuscular injection and slowly accumulated in blood. L-368,899 was 40 times more selective for OXTR than vasopressin 1a receptors and bound to the coyote OXTR with an affinity of 12 nM. These features of L-368,899 support its utility in future studies to probe the oxytocin system of coyotes.

Omega-3 long chain polyunsaturated fatty acids as a potential treatment for reducing dysmenorrhoea pain: Systematic literature review and meta-analysis.

AIM: This systematic literature review with meta-analysis aimed to determine the effect of omega-3 long chain polyunsaturated fatty acids on prostaglandin levels and pain severity in women with dysmenorrhoea and identify adverse side effects. METHODS: A literature search was conducted in Embase, Scopus, Web of Science, MEDLINE complete, CINAHL and AMED databases (PROSPERO CRD42022340371). Included studies provided omega-3 long chain polyunsaturated fatty acids compared to a control in women with dysmenorrhoea and reported pain and/or prostaglandin levels. A random effects meta-analysis with Cohen's d effect size (95% confidence interval) was performed in SPPS for studies that reported pain outcomes. Study quality was assessed using the Academy of Nutrition and Dietetics Quality Criteria Checklist. RESULTS: Twelve studies (n = 881 dysmenorrhoeal women) of predominantly neutral quality (83%) were included that provided daily supplementation of 300-1800 mg omega-3 long chain polyunsaturated fatty acids over 2 or 3 months. Meta-analysis (n = 8 studies) showed a large effect of omega-3 long chain polyunsaturated fatty acids (d = -1.020, 95% confidence interval -1.53 to -0.51) at reducing dysmenorrhoea pain. No studies measured prostaglandin levels, 86% of studies measuring analgesic use showed a reduction with omega-3 long chain polyunsaturated fatty acids and few studies reported mild adverse side effects in individual participants. CONCLUSIONS: Findings suggest that daily supplementation of 300-1800 mg omega-3 long chain polyunsaturated fatty acids over 2-3 months are generally well tolerated and reduces pain and analgesic use in women with dysmenorrhoea. However, the neutral quality of research is limited by methodological issues and the mechanism of action remains to be determined.

Can a traditional partner preference test quantify monogamous behavior in captive coyotes?

Social monogamy is a unique social system exhibited by only 3-5% of mammalian taxa; however, all wild canid species exhibit this social system. Despite the high prevalence of social monogamy among canids, little is known about how they form selective social attachment relationships among non-kin. Thus, we aimed to quantify monogamous behavior in a highly ubiquitous canid, the coyote (Canis latrans). We adapted the three-chambered partner preference test, which was originally developed for prairie voles (Microtus ochrogaster), to assess social preference in mated pairs of captive coyotes at the USDA Predator Research Facility. We quantified monogamy-related behaviors, such as time spent in spatial proximity to a pair-mate versus a stranger. Our behavioral ethogram also included visual seeking, olfactory investigations, ears down, scent marking, and affiliative behavior. Test subjects showed significantly greater affiliative behavior toward their partner than toward a stranger. However, there was extremely high variability both within and between coyote pairs across behavioral measures. These data suggest the need for larger sample sizes when working with species with high individual variability, as well as the need for species- and facility-specific modifications to this testing paradigm and/or ethogram to better adapt it from its laboratory and rodent-based origins.

Disease Surveillance of Cougars (Puma concolor) in Utah, USA.

Adult, free-ranging cougars (Puma concolor) were sampled in three regions of Utah, US, from 2018 to 2021. A total of 68% (23/34) of the sampled cougars had antibodies to feline parvovirus, 15% (5/33) to canine distemper virus, 18% (6/34) to calicivirus, and 22% (8/37) to Yersinia pestis. Forty-one percent (13/32) had IgG antibodies to Toxoplasma gondii and 6% (2/33) to feline immunodeficiency virus, and 3% (1/32) were positive for Dirofilaria immitis (heartworm) antigen. All were seronegative for Toxoplasma gondii IgM, feline enteric coronavirus, SARS-CoV-2, feline leukemia virus, feline herpesvirus, and Francisella tularensis. Tapeworms and Toxascaris leonina eggs were detected in the feces. The disease exposures detected were similar to what has been reported from cougar populations in other western US states, and the current level of exposures is unlikely to have a negative impact on the state's population.

Ecological factors shape quantitative decision-making in coyotes.

Much research has focused on the development and evolution of cognition in the realm of numerical knowledge in human and nonhuman animals but often fails to take into account ecological realities that, over time, may influence and constrain cognitive abilities in real-life decision-making. Cognitive abilities such as enumerating and timing are central to many psychological and ecological models of behavior, yet our knowledge of how these are affected by environmental fluctuations remains incomplete. Our research bridges the gap between basic cognitive research and ecological decision-making. We used coyotes (Canis latrans) as a model animal system to study decision-making about smaller, more proximal food rewards and larger, more distant food rewards; we tested animals across their four reproductive cycle phases to examine effects of ecological factors such as breeding status and environmental risk on quantitative performance. Results show that coyotes, similar to other species, spatially discount food rewards while foraging. The degree to which coyotes were sensitive to the risk of obtaining the larger food reward, however, depended on the season in which they completed the foraging task, the presence of unfamiliar humans (i.e., risk), and the presence of conspecifics. Importantly, our results support that seasonal variations drive many differences in nonhuman animal behavior and cognition (e.g., hibernation, breeding, food resource availability). Further, it may be useful in the future to extend this work to humans because seasons may influence human cognition as well, and this remains unexplored in the realms of enumeration, timing, and spatial thinking.

Scavenging vs hunting affects behavioral traits of an opportunistic carnivore.

Background: Human-induced changes to ecosystems transform the availability of resources to predators, including altering prey populations and increasing access to anthropogenic foods. Opportunistic predators are likely to respond to altered food resources by changing the proportion of food they hunt versus scavenge. These shifts in foraging behavior will affect species interactions through multiple pathways, including by changing other aspects of predator behavior such as boldness, innovation, and social structure. Methods: To understand how foraging behavior impacts predator behavior, we conducted a controlled experiment to simulate hunting by introducing a prey model to captive coyotes (Canis latrans) and compared their behavior to coyotes that continued to scavenge over one year. We used focal observations to construct behavioral budgets, and conducted novel object, puzzle box, and conspecific tests to evaluate boldness, innovation, and response to conspecifics. Results: We documented increased time spent resting by hunting coyotes paired with decreased time spent active. Hunting coyotes increased boldness and persistence but there were no changes in innovation. Our results illustrate how foraging behavior can impact other aspects of behavior, with potential ecological consequences to predator ecology, predator-prey dynamics, and human-wildlife conflict; however, the captive nature of our study limits specific conclusions related to wild predators. We conclude that human-induced behavioral changes could have cascading ecological implications that are not fully understood.

Beating the heat: ecology of desert bobcats.

BACKGROUND: Relative to temperate regions, little is known about bobcats (Lynx rufus) in the Sonoran Desert portion of their range, in part due to the difficulty of sampling an elusive carnivore in harsh desert environments. Here, we quantify habitat selection and evaluate diet of bobcats at Kofa National Wildlife Refuge, Arizona, USA, using multiple sampling techniques including GPS telemetry, camera traps, and DNA metabarcoding. RESULTS: Home ranges during the hot season were smaller than during the cool season. Camera trapping failed to yield a high enough detection rate to identify habitat occupancy trends but third-order resource selection from GPS-collar data showed a preference for higher elevations and rugged terrain at lower elevations. Diet composition consisted of a diverse range of available small prey items, including a higher frequency of avian prey than previously observed in bobcats. CONCLUSIONS: Desert bobcats in our study maintained smaller home ranges and primarily consumed smaller prey than their more northern relatives. This study illustrates the benefit of employing multiple, complementary sampling methods to understand the ecology of elusive species.

Cognitive flexibility and aging in coyotes (Canis latrans).

Cognitive flexibility evolves in species that live in complex and dynamic social systems and habitats and may enable species to better cope with anthropogenic habitat modification. Aging may also impact the cognitive abilities of canids. Coyotes (Canis latrans) and domestic dogs (Canis lupus familiaris) differ markedly in their social and trophic ecology but have both been highly successful in adapting to human-modified ecosystems. Aging dogs develop a form of dementia that mirrors Alzheimer's disease in humans, but it is unknown whether similar cognitive deficits develop with age in coyotes and other wild canids. In this study, we modified a spatial serial reversal-learning test that was sensitive to cognitive aging in pet dogs to test cognitive flexibility in captive coyotes. We also performed a second experiment using a color discrimination task to test for flexible rule learning. A total of 19 of 20 coyotes demonstrated the ability to track shifts in spatial reward contingencies and learned to rapidly complete reversals by using a win-stay, lose-shift strategy. In addition, coyotes inhibited prepotent win-stay choices to acquire the color discrimination task. These findings suggest that behavioral flexibility may help coyotes to detect and respond appropriately to both rapid fluctuations and gradual changes in ecological conditions. Performance did not differ between coyotes and previously tested dogs, but similar to dogs, behavioral flexibility declined with age in adult coyotes. Thus, cognitive decline and flexibility may be conserved among canines pending additional studies on other Canis species. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Recolonizing carnivores: Is cougar predation behaviorally mediated by bears?

Conservation and management efforts have resulted in population increases and range expansions for some apex predators, potentially changing trophic cascades and foraging behavior. Changes in sympatric carnivore and dominant scavenger populations provide opportunities to assess how carnivores affect one another. Cougars (Puma concolor) were the apex predator in the Great Basin of Nevada, USA, for over 80 years. Black bears (Ursus americanus) have recently recolonized the area and are known to heavily scavenge on cougar kills. To evaluate the impacts of sympatric, recolonizing bears on cougar foraging behavior in the Great Basin, we investigated kill sites of 31 cougars between 2009 and 2017 across a range of bear densities. We modeled the variation in feeding bout duration (number of nights spent feeding on a prey item) and the proportion of primary prey, mule deer (Odocoileus hemionus), in cougar diets using mixed-effects models. We found that feeding bout duration was driven primarily by the size of the prey item being consumed, local bear density, and the presence of dependent kittens. The proportion of mule deer in cougar diet across all study areas declined over time, was lower for male cougars, increased with the presence of dependent kittens, and increased with higher bear densities. In sites with feral horses (Equus ferus), a novel large prey, cougar consumption of feral horses increased over time. Our results suggest that higher bear densities over time may reduce cougar feeding bout durations and influence the prey selection trade-off for cougars when alternative, but more dangerous, large prey are available. Shifts in foraging behavior in multicarnivore systems can have cascading effects on prey selection. This study highlights the importance of measuring the impacts of sympatric apex predators and dominant scavengers on a shared resource base, providing a foundation for monitoring dynamic multipredator/scavenger systems.

The evolutionary consequences of human-wildlife conflict in cities.

Human-wildlife interactions, including human-wildlife conflict, are increasingly common as expanding urbanization worldwide creates more opportunities for people to encounter wildlife. Wildlife-vehicle collisions, zoonotic disease transmission, property damage, and physical attacks to people or their pets have negative consequences for both people and wildlife, underscoring the need for comprehensive strategies that mitigate and prevent conflict altogether. Management techniques often aim to deter, relocate, or remove individual organisms, all of which may present a significant selective force in both urban and nonurban systems. Management-induced selection may significantly affect the adaptive or nonadaptive evolutionary processes of urban populations, yet few studies explicate the links among conflict, wildlife management, and urban evolution. Moreover, the intensity of conflict management can vary considerably by taxon, public perception, policy, religious and cultural beliefs, and geographic region, which underscores the complexity of developing flexible tools to reduce conflict. Here, we present a cross-disciplinary perspective that integrates human-wildlife conflict, wildlife management, and urban evolution to address how social-ecological processes drive wildlife adaptation in cities. We emphasize that variance in implemented management actions shapes the strength and rate of phenotypic and evolutionary change. We also consider how specific management strategies either promote genetic or plastic changes, and how leveraging those biological inferences could help optimize management actions while minimizing conflict. Investigating human-wildlife conflict as an evolutionary phenomenon may provide insights into how conflict arises and how management plays a critical role in shaping urban wildlife phenotypes.

Variation in reversal learning by three generalist mesocarnivores.

Urbanization imposes novel challenges for wildlife, but also provides new opportunities for exploitation. Generalist species are commonly found in urban habitats, but the cognitive mechanisms facilitating their successful behavioral adaptations and exploitations are largely under-investigated. Cognitive flexibility is thought to enable generalists to be more plastic in their behavior, thereby increasing their adaptability to a variety of environments, including urban habitats. Yet direct measures of cognitive flexibility across urban wildlife are lacking. We used a classic reversal-learning paradigm to investigate the cognitive flexibility of three generalist mesocarnivores commonly found in urban habitats: striped skunks (Mephitis mephitis), raccoons (Procyon lotor), and coyotes (Canis latrans). We developed an automated device and testing protocol that allowed us to administer tests of reversal learning in captivity without extensive training or experimenter involvement. Although most subjects were able to rapidly form and reverse learned associations, we found moderate variation in performance and behavior during trials. Most notably, we observed heightened neophobia and a lack of habituation expressed by coyotes. We discuss the implications of such differences among generalists with regard to urban adaptation and we identify goals for future research. This study is an important step in investigating the relationships between cognition, generalism, and urban adaptation.

Linking Human Perceptions and Spotted Hyena Behavior in Urban Areas of Ethiopia.

Humans have shaped carnivore behavior since at least the Middle Paleolithic period, about 42,000 years ago. In more recent times, spotted hyenas (Crocuta crocuta) in Ethiopia have adapted to living in urban areas, while humans have adapted to living with hyenas. Yet, relationships between coexisting humans and carnivores are rarely addressed beyond mitigating conflicts. We provided a case study for how to broadly think about coexistence and how to study it when measuring if humans and carnivores affect one another. We collected data in four Ethiopian cities: Mekelle, Harar, Addis Ababa, and Arba Minch. We held focus groups and key informant interviews that incorporated feedback from 163 people, representing a wide array of religious, economic, and educational backgrounds. We also determined how many hyenas resided in these cities, hyena behavioral responses to humans using a flight initiation test, and problem-solving abilities via puzzle box trials. We found that in three of the cities, hyenas and humans coexist at high densities and frequently encounter each other. While all participants recognized the importance of hyenas as scavengers to maintain a clean environment, there was pronounced variation in cultural perspectives across cities. For example, while the people of Harar revere hyenas in spiritual terms, in Arba Minch hyenas were regarded as nuisance animals. Hyenas were universally respected as a formidable predator across cities but reports of attacks on livestock and humans were few. Flight initiation tests revealed hyenas fled at significantly closer distances in Harar and Addis Ababa than in Mekelle. Hyenas succeeded at solving a puzzle box in Harar but not in Mekelle. These variable behavior in hyenas correlated to different human perceptions. Our case study results suggest that the hyena-human dynamic is highly variable across these locations. We conclude by exploring the implications of these findings for how humans and hyenas can shape one another's behavior. Developing studies to link human perceptions and animal behavior could advance wildlife conservation, especially in urban areas.

Interactions with humans shape coyote responses to hazing.

Medium and large carnivores coexist with people in urban areas globally, occasionally resulting in negative interactions that prompt questions about how to reduce human-wildlife conflict. Hazing, i.e., scaring wildlife, is frequently promoted as an important non-lethal means for urbanites to reduce conflict but there is limited scientific evidence for its efficacy. We used a population of captive coyotes (Canis latrans) to simulate urban human-coyote interactions and subsequent effects of hazing on coyote behavior. Past experiences with humans significantly affected the number of times a coyote approached a human to necessitate hazing. Coyotes that had been hand fed by adults had to be more frequently hazed than coyotes with other or no past experiences with adults. Past experience with children had no impact on the number of hazing events. The number of times a coyote approached an adult or child was reduced across days based on the accumulative number of times hazed, suggesting coyotes learn to avoid behaviors warranting hazing and that this could be used as a non-lethal management tool. However, prior experience and whether the interaction is with an adult or child can alter the outcomes of hazing and must be considered in determining the efficacy of hazing programs.

Persistence and conspecific observations improve problem-solving abilities of coyotes.

Social learning has important ecological and evolutionary consequences but the role of certain factors, such as social rank, neophobia (i.e., avoidance of novel stimuli), persistence, and task-reward association, remain less understood. We examined the role of these factors in social learning by captive coyotes (Canis latrans) via three studies. Study 1 involved individual animals and eliminated object neophobia by familiarizing the subjects to the testing apparatus prior to testing. Studies 2 and 3 used mated pairs to assess social rank, and included object neophobia, but differed in that study 3 decoupled the food reward from the testing apparatus (i.e., altered task-reward association). For all three studies, we compared performance between coyotes that received a demonstration from a conspecific to control animals with no demonstration prior to testing. Coyotes displayed social learning during study 1; coyotes with a demonstrator were faster and more successful at solving the puzzle box but did not necessarily use the same modality as that observed to be successful. In study 2, there was no difference in success between treatment groups but this is likely because only one coyote within each pair was successful so successful coyote results were masked by their unsuccessful mate. In study 3, there was no difference in success between treatment groups; only two coyotes, both dominant, hand-reared males with demonstrators were able to perform the task. However, coyotes with a demonstrator were less neophobic, measured as latency to approach the object, and more persistent, measured as time spent working on the apparatus. Social rank was the best predictor of neophobia and persistence and was also retained in the best model for time to eat inside the apparatus, a post-trial measurement of object neophobia. These results suggest coyotes are capable of social learning for novel tasks but social rank, neophobia, and persistence influence their social-learning capabilities. This study contributes to understanding the mechanisms underlying how animals gain information about their environment.

Space Use and Movement of Urban Bobcats.

Global urbanization is rapidly changing the landscape for wildlife species that must learn to persist in declining wild spacing, adapt, or risk extinction. Many mesopredators have successfully exploited urban niches, and research on these species in an urban setting offers insights into the traits that facilitate their success. In this study, we examined space use and activity patterns from GPS-collared bobcats (Lynx rufus) in the Dallas-Fort Worth metroplex, Texas, USA. We found that bobcats select for natural/agricultural features, creeks, and water ways and there is greater home-range overlap in these habitats. They avoid roads and are less likely to have home-range overlap in habitats with more roads. Home-range size is relatively small and overlap relatively high, with older animals showing both greater home-range size and overlap. Simultaneous locations suggest bobcats are neither avoiding nor attracted to one another, despite the high overlap across home ranges. Finally, bobcats are active at all times of day and night. These results suggest that access to natural features and behavioral plasticity may enable bobcats to live in highly developed landscapes.

The intrepid urban coyote: a comparison of bold and exploratory behavior in coyotes from urban and rural environments.

Coyotes (Canis latrans) are highly adaptable, medium-sized carnivores that now inhabit nearly every large city in the United States and Canada. To help understand how coyotes have adapted to living in urban environments, we compared two ecologically and evolutionarily important behavioral traits (i.e., bold-shy and exploration-avoidance behavior) in two contrasting environments (i.e., rural and urban). Boldness is an individual's reaction to a risky situation and exploration is an individual's willingness to explore novel situations. Our results from both tests indicate that urban coyotes are bolder and more exploratory than rural coyotes and that within both populations there are individuals that vary across both spectrums. Bolder behavior in urban coyotes emerged over several decades and we speculate on possible processes (e.g., learning and selection) and site differences that could be playing a role in this behavioral adaptation. We hypothesize that an important factor is how people treat coyotes; in the rural area coyotes were regularly persecuted whereas in the urban area coyotes were rarely persecuted and sometimes positively rewarded to be in close proximity of people. Negative consequences of this behavioral adaptation are coyotes that become bold enough to occasionally prey on pets or attack humans.

How Behavior of Nontarget Species Affects Perceived Accuracy of Scat Detection Dog Surveys.

Detection dogs, specially trained domestic dogs (Canis familiaris), have become a valuable, noninvasive, conservation tool because they remove the dependence of attracting species to a particular location. Further, detection dogs locate samples independent of appearance, composition, or visibility allowing researchers to collect large sets of unbiased samples that can be used in complex ecological queries. One question not fully addressed is why samples from nontarget species are inadvertently collected during detection dog surveys. While a common explanation has been incomplete handler or dog training, our study aimed to explore alternative explanations. Our trials demonstrate that a scat's genetic profile can be altered by interactions of nontarget species with target scat via urine-marking, coprophagy, and moving scats with their mouths, all pathways to contamination by nontarget species' DNA. Because detection dogs are trained to locate odor independent of masking, the collection of samples with a mixed olfactory profile (target and nontarget) is possible. These scats will likely have characteristics of target species' scats and are therefore only discovered faulty once genetic results indicate a nontarget species. While the collection of nontarget scats will not impact research conclusions so long as samples are DNA tested, we suggest ways to minimize their collection and associated costs.

Spatial processes decouple management from objectives in a heterogeneous landscape: predator control as a case study.

Predator control is often implemented with the intent of disrupting top-down regulation in sensitive prey populations. However, ambiguity surrounding the efficacy of predator management, as well as the strength of top-down effects of predators in general, is often exacerbated by the spatially implicit analytical approaches used in assessing data with explicit spatial structure. Here, we highlight the importance of considering spatial context in the case of a predator control study in south-central Utah. We assessed the spatial match between aerial removal risk in coyotes (Canis latrans) and mule deer (Odocoileus hemionus) resource selection during parturition using a spatially explicit, multi-level Bayesian model. With our model, we were able to evaluate spatial congruence between management action (i.e., coyote removal) and objective (i.e., parturient deer site selection) at two distinct scales: the level of the management unit and the individual coyote removal. In the case of the former, our results indicated substantial spatial heterogeneity in expected congruence between removal risk and parturient deer site selection across large areas, and is a reflection of logistical constraints acting on the management strategy and differences in space use between the two species. At the level of the individual removal, we demonstrated that the potential management benefits of a removed coyote were highly variable across all individuals removed and in many cases, spatially distinct from parturient deer resource selection. Our methods and results provide a means of evaluating where we might anticipate an impact of predator control, while emphasizing the need to weight individual removals based on spatial proximity to management objectives in any assessment of large-scale predator control. Although we highlight the importance of spatial context in assessments of predator control strategy, we believe our methods are readily generalizable in any management or large-scale experimental framework where spatial context is likely an important driver of outcomes.

Parental habituation to human disturbance over time reduces fear of humans in coyote offspring.

A fundamental tenet of maternal effects assumes that maternal variance over time should have discordant consequences for offspring traits across litters. Yet, seldom are parents observed across multiple reproductive bouts, with few studies considering anthropogenic disturbances as an ecological driver of maternal effects. We observed captive coyote (Canis latrans) pairs over two successive litters to determine whether among-litter differences in behavior (i.e., risk-taking) and hormones (i.e., cortisol and testosterone) corresponded with parental plasticity in habituation. Thus, we explicitly test the hypothesis that accumulating experiences of anthropogenic disturbance reduces parental fear across reproductive bouts, which should have disparate phenotypic consequences for first- and second-litter offspring. To quantify risk-taking behavior, we used foraging assays from 5-15 weeks of age with a human observer present as a proxy for human disturbance. At 5, 10, and 15 weeks of age, we collected shaved hair to quantify pup hormone levels. We then used a quantitative genetic approach to estimate heritability, repeatability, and between-trait correlations. We found that parents were riskier (i.e., foraged more frequently) with their second versus first litters, supporting our prediction that parents become increasingly habituated over time. Second-litter pups were also less risk-averse than their first-litter siblings. Heritability for all traits did not differ from zero (0.001-0.018); however, we found moderate support for repeatability in all observed traits (r = 0.085-0.421). Lastly, we found evidence of positive phenotypic and cohort correlations among pup traits, implying that cohort identity (i.e., common environment) contributes to the development of phenotypic syndromes in coyote pups. Our results suggest that parental habituation may be an ecological cue for offspring to reduce their fear response, thus emphasizing the role of parental plasticity in shaping their pups' behavioral and hormonal responses toward humans.

Studies of wolf x coyote hybridization via artificial insemination.

Following the production of western gray wolf (Canis lupus) x western coyote (Canis latrans) hybrids via artificial insemination (AI), the present article documents that the hybrids survived in captivity for at least 4 years and successfully bred with each other. It further reports that backcrossing one of the hybrids to a male gray wolf by AI also resulted in the birth of live pups that have survived for at least 10 months. All male hybrids (F1 and F2) produced sperm by about 10 months of age, and sperm quality of the F1 males fell within the fertile range for domestic dogs, but sperm motility and morphology, in particular, were low in F2 males at 10 months but improved in samples taken at 22 months of age. These studies are relevant to a long-standing controversy about the identity of the red wolf (Canis rufus), the existence of a proposed new species (Canis lycaon) of gray wolf, and to the role of hybridization in mammalian evolution.