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Urbanisation and habitat loss are major threats to wildlife populations globally. Understanding how species respond to anthropogenic changes is therefore crucial to mitigating threats and developing conservation management strategies. We examined the habitat use of five fishing cats (Prionailurus viverrinus) in Sri Lanka’s capital city, Colombo, a densely urbanised landscape with a mosaic of wetland habitats, cultivated areas, and altered open spaces. We investigated: (1) to what extent all five cats used human-impacted versus natural wetland habitat; (2) whether there were behavioural shifts to avoid human activity throughout the diel cycle; (3) the home range sizes of two resident females and one resident male, and the extent of the area used by the two translocated males; (4) whether the two translocated males would survive introduction to a new urban environment. We monitored the fishing cats for 637 days (mean = 127) and collected a total of 2278 GPS (5-h interval) collar locations. We found that all five individuals used highly urbanised areas more than we expected. Home range sizes of the three residents were smaller than fishing cat home ranges in less disturbed landscapes. Though our sample size was small, our findings suggest that fishing cats use urbanised areas in Colombo, particularly at night, likely to avoid daytime human activity. Further comprehensive ecological study is needed to explore the aspects of fishing cat ecology that facilitate their persistence, and aid in their conservation across increasingly urbanised areas.

Anellovirus infections are highly prevalent in mammals, however, prior to this study only a handful of anellovirus genomes had been identified in members of the Felidae family. Here we characterise anelloviruses in pumas (Puma concolor), bobcats (Lynx rufus), Canada lynx (Lynx canadensis), caracals (Caracal caracal) and domestic cats (Felis catus). The complete anellovirus genomes (n = 220) recovered from 149 individuals were diverse. ORF1 protein sequence similarity network analysis coupled with phylogenetic analysis, revealed two distinct clusters that are populated by felid-derived anellovirus sequences, a pattern mirroring that observed for the porcine anelloviruses. Of the two-felid dominant anellovirus groups, one includes sequences from bobcats, pumas, domestic cats and an ocelot, and the other includes sequences from caracals, Canada lynx, domestic cats and pumas. Coinfections of diverse anelloviruses appear to be common among the felids. Evidence of recombination, both within and between felid-specific anellovirus groups, supports a long coevolution history between host and virus. ispartof: VIROLOGY vol:562 pages:176-189 ispartof: location:United States status: published

Spatially heterogeneous landscape factors such as urbanisation can have substantial effects on the severity and spread of wildlife diseases. However, research linking patterns of pathogen transmission to landscape features remains rare. Using a combination of phylogeographic and machine learning approaches, we tested the influence of landscape and host factors on feline immunodeficiency virus (FIVLru) genetic variation and spread among bobcats (Lynx rufus) sampled from coastal southern California. We found evidence for increased rates of FIVLru lineage spread through areas of higher vegetation density. Furthermore, single-nucleotide polymorphism (SNP) variation among FIVLru sequences was associated with host genetic distances and geographic location, with FIVLru genetic discontinuities precisely correlating with known urban barriers to host dispersal. An effect of forest land cover on FIVLru SNP variation was likely attributable to host population structure and differences in forest land cover between different populations. Taken together, these results suggest that the spread of FIVLru is constrained by large-scale urban barriers to host movement. Although urbanisation at fine spatial scales did not appear to directly influence virus transmission or spread, we found evidence that viruses transmit and spread more quickly through areas containing higher proportions of natural habitat. These multiple lines of evidence demonstrate how urbanisation can change patterns of contact-dependent pathogen transmission and provide insights into how continued urban development may influence the incidence and management of wildlife disease.

We used mitochondrial sequences and nuclear microsatellites to investigate population structure of gray foxes (Urocyon cinereoargenteus) and the evolutionary origins of the endemic island fox (Urocyon littoralis), which first appeared in the northern Channel Islands

Polyomaviruses are oncogenic viruses that are generally thought to have co-evolved with their hosts. While primate and rodent polyomaviruses are increasingly well-studied, less is known about polyomaviruses that infect other mammals. In an effort to gain insight into polyomaviruses associated with carnivores, we surveyed fecal samples collected in the USA from bobcats (Lynx rufus), pumas (Puma concolor), Canada lynxes (Lynx canadensis), and grizzly bears (Ursus arctos). Using a viral metagenomic approach, we identified six novel polyomavirus genomes. Surprisingly, four of the six genomes showed a phylogenetic relationship to polyomaviruses found in prey animals. These included a putative rabbit polyomavirus from a bobcat fecal sample and two possible deer-trophic polyomaviruses from Canada lynx feces. One polyomavirus found in a grizzly bear sample was found to be phylogenetically distant from previously identified polyomaviruses. Further analysis of the grizzly bear fecal sample showed that it contained anelloviruses that are known to infect pigs, suggesting that the bear might have preyed on a wild or domestic pig. Interestingly, a polyomavirus genome identified in a puma fecal sample was found to be closely related both to raccoon polyomavirus 1 and to Lyon-IARC polyomavirus, the latter of which was originally identified in human saliva and skin swab specimens but has since been found in samples from domestic cats (Felis catus).

In human-dominated landscapes, habitat fragmentation and barriers to movement can interrupt gene flow. While often considered at a local extent, regional analyses are also needed to reveal broader landscape-mediated population processes. To explore the relationship between patterns of gene flow and fragmentation resulting from urbanization across southern California, we used the bobcat as an indicator species. We assembled data for a landscape level genetic analysis across southern California from both archived and new samples, including two northern Californian populations for comparison, to identify local and regional areas affected by isolation. Our regional analyses focused on a dataset of 19 microsatellite loci for 118 individuals and a dataset of 422 individuals genotyped at 11 loci. We examined population genetic structure and examined how pairwise genetic distance of all population clusters aligned with geographic distance. We employed a landscape genetic analysis based on resistance to determine which features of the landscape likely play a role in determining the patterns of genetic structure we observed. Study populations generally exhibited a pattern of isolation by distance and localized areas of genetic isolation. The landscape genetic analysis suggested that, in southern California, these patterns are driven by overall landscape permeability. Although local studies are key to examining the effects of urbanization and habitat fragmentation on populations, we demonstrate the importance of combining local and regional analyses for wide-ranging species to understand and maintain connectivity at local scales, while also improving and establishing sustainable linkages to habitats at the regional scale.

Urbanisation radically changes habitats and alters available resources. Populations of large, highly mobile species are often extirpated at the urban-wildland interface, while species like mesocarnivores may thrive by capitalising on changes in prey abundance. We investigated the diet of the caracal (Caracal caracal), a medium-sized felid inhabiting patchy natural habitat isolated within the dense urban matrix of South Africa’s second largest city, Cape Town. We systematically integrated two classic dietary methods (scat and GPS clusters) by accounting for gut transit times. As part of a larger caracal ecology study, we GPS-collared 26 individuals over a two-year period (2014–2016) to generate coarse (3-hour) and fine-scale (20-minute) GPS movement data. Using the movement data, we investigated 677 GPS-clusters for prey remains. We collected 654 scats, half of which were found at GPS-clusters and were linked with the individual sampled. By systematically correcting for a range of gut transit times, we determined whether scat at cluster sites was from the same or an earlier feeding event, thereby increasing the overall detection of feeding events by > 50%. Avian prey dominated GPS cluster findings while micromammals were overwhelmingly represented in scat. Although > 40% of feeding events occurred within 200 meters of the urban edge, caracals largely preyed on native species. Our findings have implications for understanding the ability of some species to persist in the face of rapid environmental change, human-wildlife conflict, pathogen transmission, and bioaccumulation of pesticides. Further, this approach could be incorporated into studies that estimate foraging-explicit resource selection and habitat preference.

Wildlife populations are increasingly challenged by human activities that disrupt landscape connectivity, animal movement, population dynamics and population persistence. Yet modified habitats may provide resource subsidies for generalist species resulting in increased selection of disturbed areas. Understanding how species adjust their space use and activity in human-modified landscapes is fundamental to conserving wildlife populations globally. To test three competing hypotheses explaining spatiotemporal responses to human activity, we investigated coarse (3 hour)- and fine (20 minute)- scale habitat selection, activity patterns, and measured home ranges in a human-dominated landscape. We GPS-collared 25 adults and subadult caracals (Caracal caracal) from three contiguous subpopulations in urban and wildland-dominated regions. Caracals in the wildland-dominated region avoided proximity to urban areas while caracals in the urban-dominated region selected for close proximity to urban areas. Selection responses for vegetative cover, freshwater, vineyards, the coast, and burned areas was dependent on age class, diel period, and whether individuals were in close proximity to urban areas. Large home ranges demonstrated habitat selection results were not a result of limited landscape use. Caracals did not shift their temporal activity to avoid humans, but selected for closer proximity to urban areas at night, suggesting they seek spatial refugia during the day. Behavioral plasticity whereby individuals seek out spatial refugia promotes the coexistence of this carnivore in a human-dominated landscape. These data are essential to our understanding of what is viable wildlife habitat to a generalist mesocarnivore and provide opportunities to guide land acquisition and conservation by local management agencies.

Smacoviruses are small circular single-stranded DNA viruses that appear to be prevalent in faeces of a range of animals and have also been found in a few insect species. In this study, we report the first viral genomes from faeces of free-roaming wild felids on two continents. Two smacoviruses were recovered from the faeces of two North American bobcats (Lynx rufus), and one was recovered from an African lion (Panthera leo). All three genomes are genetically different, sharing 59-69% genome-wide sequence identity to other smacoviruses. These are the first full smacovirus genome sequences associated with a large top-end feline predator, and their presence in these samples suggests that feline faeces are a natural niche for the organisms that these viruses infect. Electronic supplementary material The online version of this article (10.1007/s00705-019-04329-3) contains supplementary material, which is available to authorized users.

Anticoagulant rodenticides (ARs) are used worldwide to control rodent populations. ARs bioaccumulate across trophic levels and threaten non-target wildlife. We investigated the prevalence of AR exposure in seven predator species in the rapidly developing Greater Cape Town region of South Africa – a mosaic of natural, urban, and agricultural areas within a global biodiversity hotspot. We focused sampling on caracals (Caracal caracal, n = 28) as part of a larger caracal ecology study, but also opportunistically sampled Cape Clawless otters (Aonyx capensis, n = 9), large-spotted genets (Genetta tigrina, n = 4), honey badger (Mellivora capensis, n = 1), water mongoose (Atilax paludinosus, n = 1), small gray mongoose (Galerella pulverulenta, n = 1), and Cape Eagle owl (Bubo capensis, n = 1). We tested livers from all species, and blood from ten caracals, for eight AR compounds to assess prevalence and amount of exposure for each compound. We used generalized linear models to test spatial, demographic, and seasonal risk factors for ten measures of AR exposure in caracals. We detected at least one of the four most toxic AR compounds in six species. Exposure was high for caracals (92%) and all species combined (81%). For caracals, proximity to vineyards was the most important AR exposure risk factor. Vineyards in Cape Town do not use ARs to protect their vines but do host commercial hospitality structures where ARs are used. Vineyards may thus link caracals that forage within vineyards to the rat poisons used in and around their commercial structures. Residue levels were unexpected in large-spotted genets and Cape Clawless otters, suggesting invertebrate vectors. ARs may present a cryptic threat to populations already vulnerable to increasing habitat loss, vehicle collisions, poachers and fire. Targeted mitigation should include a mix of environmentally responsible policies that reduce AR use, particularly in areas near wildlife habitat.

Background: Wildlife populations are increasingly challenged by human activities that disrupt landscape connectivity and animal movement, and thus population dynamics and persistence. Yet modified habitats may provide resource subsidies for generalist species resulting in increased selection of disturbed areas. Understanding how species adjust their space use and activity in human-modified landscapes is increasingly critical to conserving wildlife populations globally. Methods: To test three competing hypotheses explaining spatiotemporal responses to human activity, we GPS-collared 26 individual adults and subadults from three contiguous subpopulations in wildland and urban-dominated regions. We investigated coarse (3 hour)- and fine (20 minute)- scale movement-explicit habitat selection using step selection functions and tested for increased nocturnality of caracals (Caracal caracal) in a human-dominated landscape in South Africa. Results: Caracals in a wildland-dominated landscape were on average > 8.0 km from the urban-natural boundary and strongly selected for proximity to the coastline. Caracals in the urban-dominated landscape were on average Conclusions: Behavioral plasticity whereby individuals seek out spatial, but not temporal refugia, appears to promote the coexistence of this generalist carnivore in a human-dominated landscape. Both age class and degree of exposure to anthropogenic activities influence habitat selection, while some individuals have learned to successfully exploit the riskiest areas at night. These data are essential to our understanding of what is viable wildlife habitat in this rapidly urbanizing landscape and provide opportunities to guide land acquisition and conservation guidelines by local management agencies.

Background Wild carnivores living alongside humans and domestic animals are vulnerable to changes in the infectious disease dynamics in their populations. The aims of this study were to determine the prevalence and diversity of selected tick-borne pathogens (TBPs) of veterinary and/or zoonotic concern in wild populations of caracals (Caracal caracal) occurring in human-modified landscapes in South Africa. Using molecular techniques, we screened 57 caracal blood samples for infection by rickettsial bacteria and piroplasms in three regions of South Africa: rangeland in the Central Karoo (n = 27) and Namaqualand (n = 14) as well as the urban edge of the Cape Peninsula (n = 16) of South Africa. To characterise pathogen identity, we sequenced the 18S rRNA and 16S rRNA genes from positive samples and analysed sequences within a phylogenetic framework. We also examine the diversity of potential tick vectors. Results All individuals tested were infected with at least one tick-borne pathogen. Pathogens included Hepatozoon felis, Babesia felis, Babesia leo and a potentially novel Babesia species. An Anaplasma species previously described in South African domestic dogs was also found in 88% of urban edge caracals. Higher rates of co-infection characterised urban edge caracals (81% vs 15% and 0% in the two rangeland populations), as well as a greater incidence of mixed infections. Host attached tick species include Haemaphysalis elliptica, an important pathogen vector among carnivore hosts. Conclusions This study confirms the occurrence of previously undocumented tick-borne pathogens infecting free-ranging caracals in human-modified landscapes. We identify clear differences in the pathogen profiles among our study populations and discuss the likely health costs to caracals living adjacent to urban areas.

Understanding the environmental contributors to population structure is of paramount importance for conservation in urbanized environments. We used spatially explicit models to determine genetic population structure under current and future environmental conditions across a highly fragmented, human-dominated environment in Southern California to assess the effects of natural ecological variation and urbanization. We focused on 7 common species with diverse habitat requirements, home-range sizes, and dispersal abilities. We quantified the relative roles of potential barriers, including natural environmental characteristics and an anthropogenic barrier created by a major highway, in shaping genetic variation. The ability to predict genetic variation in our models differed among species: 11-81% of intraspecific genetic variation was explained by environmental variables. Although an anthropogenically induced barrier (a major highway) severely restricted gene flow and movement at broad scales for some species, genetic variation seemed to be primarily driven by natural environmental heterogeneity at a local level. Our results show how assessing environmentally associated variation for multiple species under current and future climate conditions can help identify priority regions for maximizing population persistence under environmental change in urbanized regions.

We investigated

Landscape connectivity is essential to conserving resilient wildlife populations in the Anthropocene. Maintaining connectivity requires preserving or restoring patches of habitat, accounting for the behavioral factors that determine movement between patches and mitigating threats. We measured natural and anthropogenic features that influence movement and mortality for bobcats (Lynx rufus) in a system threatened with complete isolation by urbanization. Our overarching objective was to inform local land acquisition and restoration to maintain two last-chance wildlife corridors. We collected five-minute movement data from 36 bobcats in central California to assess features of road-crossing hotspots and model habitat selection, including functional responses to housing densities and vegetation. We collected opportunistic mortalities and assessed rat poison exposure to evaluate edge effects as acute threats. Bobcats strongly selected for natural vegetation, evident at the level of a single tree or shrub. Individuals selected low-density housing (

SUMMARYNotoedric mange, caused by obligately parasitic sarcoptiform Notoedres mites, is associated with potentially fatal dermatitis with secondary systemic disease in small mammals, felids and procyonids among others, as well as an occasional zoonosis. We describe clinical spectra in non-chiropteran hosts, review risk factors and summarize ecological and epidemiological studies. The genus is disproportionately represented on rodents. Disease in felids and procyonids ranges from very mild to death. Knowledge of the geographical distribution of the mites is highly inadequate, with focal hot spots known for Notoedres cati in domestic cats and bobcats. Predisposing genetic and immunological factors are not known, except that co-infection with other parasites and anticoagulant rodenticide toxicoses may contribute to severe disease. Treatment of individual animals is typically successful with macrocytic lactones such as selamectin, but herd or wildlife population treatment has not been undertaken. Transmission requires close contact and typically is within a host species. Notoedric mange can kill half all individuals in a population and regulate host population below non-diseased density for decades, consistent with frequency-dependent transmission or spillover from other hosts. Epidemics are increasingly identified in various hosts, suggesting global change in suitable environmental conditions or increased reporting bias.

The Publisher would like to correct the introduced formatting errors on the caption of Figure 1 and in the data in Table 2.

Urbanization is a major factor driving habitat fragmentation and connectivity loss in wildlife. However, the impacts of urbanization on connectivity can vary among species and even populations due to differences in local landscape characteristics, and our ability to detect these relationships may depend on the spatial scale at which they are measured. Bobcats (Lynx rufus) are relatively sensitive to urbanization and the status of bobcat populations is an important indicator of connectivity in urban coastal southern California. We genotyped 271 bobcats at 13,520 SNP loci to conduct a replicated landscape resistance analysis in five genetically distinct populations. We tested urban and natural factors potentially influencing individual connectivity in each population separately, as well as study-wide. Overall, landscape genomic effects were most frequently detected at the study-wide spatial scale, with urban land cover (measured as impervious surface) having negative effects and topographic roughness having positive effects on gene flow. The negative effect of urban land cover on connectivity was also evident when populations were analyzed separately despite varying substantially in spatial area and the proportion of urban development, confirming a pervasive impact of urbanization largely independent of spatial scale. The effect of urban development was strongest in one population where stream habitat had been lost to development, suggesting that riparian corridors may help mitigate reduced connectivity in urbanizing areas. Our results demonstrate the importance of replicating landscape genetic analyses across populations and considering how landscape genetic effects may vary with spatial scale and local landscape structure.

Author(s): Serieys, Laurel E. K.; Lea, Amanda; Epeldegui, Marta; Foley, Janet; Moriarty, Joanne Gale; Riley, Seth P. D.; Uittenbogaart, Christel H.; Fraser, Devaughn; Mouton, Alice; Wayne, Robert K. | Abstract: Human activities threaten wildlife with a variety of novel stressors such as exposure to toxicants. Anticoagulant rodenticides (ARs) are toxicants applied worldwide and through bioaccumulation, threaten species that prey on poisoned rodents or their predators. We studied a population of urban bobcats in southern California that declined rapidly from 2002-2005 due to notoedric mange. We first assessed prevalence of AR exposure using blood and liver samples across the population and found widespread exposure (g90%). Death associated with mange was strongly correlated with cumulative first- and second-generation AR exposure. These findings suggested that exposure to both first- and second-generation ARs were an underlying cause of the disease. We next aimed to understand the sublethal immunological and physiological effects of AR exposure in this natural population. We used two approaches: 1) we used a comprehensive suite of health assays (complete blood counts, blood chemistry assessment, and immunological profiling), and 2) we quantified AR-induced differential gene expression in blood for a subset of individuals. We found that sublethal AR exposure, primarily measured as exposure to diphacinone, is associated with hallmark indicators of generalized systemic inflammation that in persistence could promote immune dysfunction. Further, differential gene expression findings supported the results of immunological profiling. Further, a decrease in the expression of genes associated with epithelial maintenance simultaneous to a decrease in gene expression linked with ectoparasitic immune response may explain the link between AR exposure and mange vulnerability. Such indirect effects of sublethal exposure exemplify the challenge of protecting wild populations from common toxicants in human-dominated environments.

Anticoagulant rodenticides (ARs) are indiscriminate toxicants that threaten nontarget predatory and scavenger species through secondary poisoning. Accumulating evidence suggests that AR exposure may have disruptive sublethal consequences on individuals that can affect fitness. We evaluated AR-related effects on genome-wide expression patterns in a population of bobcats in southern California. We identify differential expression of genes involved in xenobiotic metabolism, endoplasmic reticulum stress response, epithelial integrity and both adaptive and innate immune function. Further, we find that differential expression of immune-related genes may be attributable to AR-related effects on leucocyte differentiation. Collectively, our results provide an unprecedented understanding of the sublethal effects of AR exposure on a wild carnivore. These findings highlight potential detrimental effects of ARs on a wide variety of species worldwide that may consume poisoned rodents and indicate the need to investigate gene expression effects of other toxicants added to natural environments by humans.

Owing to a complex history of host-parasite coevolution, lentiviruses exhibit a high degree of species specificity. Given the well-documented viral archeology of human immunodeficiency virus (HIV) emergence following human exposures to simian immunodeficiency virus (SIV), an understanding of processes that promote successful cross-species lentiviral transmissions is highly relevant. We previously reported natural cross-species transmission of a subtype of feline immunodeficiency virus, puma lentivirus A (PLVA), between bobcats ( Lynx rufus ) and mountain lions ( Puma concolor ) for a small number of animals in California and Florida. In this study, we investigate host-specific selection pressures, within-host viral fitness, and inter- versus intraspecies transmission patterns among a larger collection of PLV isolates from free-ranging bobcats and mountain lions. Analyses of proviral and viral RNA levels demonstrate that PLVA fitness is severely restricted in mountain lions compared to that in bobcats. We document evidence of diversifying selection in three of six PLVA genomes from mountain lions, but we did not detect selection among 20 PLVA isolates from bobcats. These findings support the hypothesis that PLVA is a bobcat-adapted virus which is less fit in mountain lions and under intense selection pressure in the novel host. Ancestral reconstruction of transmission events reveals that intraspecific PLVA transmission has occurred among panthers ( Puma concolor coryi ) in Florida following the initial cross-species infection from bobcats. In contrast, interspecific transmission from bobcats to mountain lions predominates in California. These findings document outcomes of cross-species lentiviral transmission events among felids that compare to the emergence of HIV from nonhuman primates. IMPORTANCE Cross-species transmission episodes can be singular, dead-end events or can result in viral replication and spread in the new species. The factors that determine which outcome will occur are complex, and the risk of new virus emergence is therefore difficult to predict. We used molecular techniques to evaluate the transmission, fitness, and adaptation of puma lentivirus A (PLVA) between bobcats and mountain lions in two geographic regions. Our findings illustrate that mountain lion exposure to PLVA is relatively common but does not routinely result in communicable infections in the new host. This is attributed to efficient species barriers that largely prevent lentiviral adaptation. However, the evolutionary capacity for lentiviruses to adapt to novel environments may ultimately overcome host restriction mechanisms over time and under certain ecological circumstances. This phenomenon provides a unique opportunity to examine cross-species transmission events leading to new lentiviral emergence.

Urbanization profoundly impacts animal populations by causing isolation, increased susceptibility to disease, and exposure to toxicants. Genetic effects include reduced effective population size, increased population substructure, and decreased adaptive potential. We investigated the influence that urbanization and a disease epizootic had on the population genetics of bobcats (Lynx rufus) distributed across a highly fragmented urban landscape. We genotyped more than 300 bobcats, sampled from 1996 to 2012, for variation at nine neutral and seven immune gene-linked microsatellite loci. We found that two freeways are significant barriers to gene flow. Further, a 3-year disease epizootic, associated with secondary anticoagulant rodenticide exposure, caused a population bottleneck that led to significant genetic differentiation between pre- and post-disease populations that was greater than that between populations separated by major freeways for >60 years. However, balancing selection acted on immune-linked loci during the epizootic, maintaining variation at functional regions. Conservation assessments need to assay loci that are potentially under selection to better preserve the adaptive potential of populations at the urban–wildland interface. Further, interconnected regions that contain appropriate habitat for wildlife will be critical to the long-term viability of animal populations in urban landscapes.

We investigated Toxoplasmosis gondii antibody seroprevalence in free-ranging caracals (Caracal caracal) in Cape Town, South Africa, from 2014 to 2017. Seropositivity was 83% (24/29), which...

Notoedric mange was responsible for a population decline of bobcats ( Lynx rufus ) in 2 Southern California counties from 2002-2006 and is now reported to affect bobcats in Northern and Southern California. With this study we document clinical laboratory and necropsy findings for bobcats with mange. Bobcats in this study included free-ranging bobcats with mange (n = 34), a control group of free-ranging bobcats without mange (n = 11), and a captive control group of bobcats without mange (n = 19). We used 2 control groups to evaluate potential anomalies due to capture stress or diet. Free-ranging healthy and mange-infected bobcats were trapped or salvaged. Animals were tested by serum biochemistry, complete blood count, urine protein and creatinine, body weight, necropsy, and assessment for anticoagulant rodenticide residues in liver tissue. Bobcats with severe mange were emaciated, dehydrated, and anemic with low serum creatinine, hyperphosphatemia, hypoglycemia, hypernatremia, and hyperchloremia, and sometimes septicemic when compared to control groups. Liver enzymes and leukocyte counts were elevated in free-ranging, recently captured bobcats whether or not they were infested with mange, suggesting capture stress. Bobcats with mange had lower levels of serum cholesterol, albumin, globulin, and total protein due to protein loss likely secondary to severe dermatopathy. Renal insufficiency was unlikely in most cases, as urine protein:creatinine ratios were within normal limits. A primary gastrointestinal loss of protein or blood was possible in a few cases, as evidenced by elevated blood urea nitrogen, anemia, intestinal parasitism, colitis, gastric hemorrhage, and melena. The prevalence of exposure to anticoagulant rodenticides was 100% (n = 15) in bobcats with mange. These findings paint a picture of debilitating, multisystemic disease with infectious and toxic contributing factors that can progress to death in individuals and potential decline in populations.

Notoedric mange in felids is a devastating disease caused by a hypersensitivity reaction to the mite Notoedres cati. The burrowing of the mite causes intense pruritis resulting in self-mutilation, secondary bacterial infection, and often death of affected felids if left untreated. Our understanding of how notoedric mange is maintained in felid populations, and the true geographic extent of infestations, has been hampered because wild felids are elusive and, thus, traditional diagnostic methods are difficult to implement. To create a noninvasive diagnostic test, we developed and validated a novel PCR assay to detect N. cati DNA in fecal samples of bobcats (Lynx rufus) and used this assay to investigate a recent outbreak of mange in northern California, United States. Although the fecal PCR assay was 100% specific and could detect as few as 1.9 mites/200 μg of feces, it had a moderate sensitivity of 52.6%, potentially due to intermittent shedding of mites in feces or fecal PCR inhibitors. In a field investigation, 12% (95% confidence interval [CI]: 0.06, 0.23) of fecal samples (n=65) collected from Rancho San Antonia County Park and Open Space Preserve in Santa Clara County, California were PCR-positive for N. cati. When this estimate was adjusted for test sensitivity, the corrected proportion for fecal samples containing N. cati was 23% (95% CI: 0.14, 0.36), suggesting widespread mange in this area. This novel PCR assay will be an important tool to assess the distribution and spread of notoedric mange in bobcats and could be validated to test other wild felids such as mountain lions (Puma concolor). The assay could also be used to detect notoedric mange in domestic cats (Felis catus), particularly feral cats, which may also suffer from mange and could represent an important contributor to mange in periurban bobcat populations.

The extinction vortex is a theoretical model describing the process by which extinction risk is elevated in small, isolated populations owing to interactions between environmental, demographic, and genetic factors. However, empirical demonstrations of these interactions have been elusive. We modelled the dynamics of a small mountain lion population isolated by anthropogenic barriers in greater Los Angeles, California, to evaluate the influence of demographic, genetic, and landscape factors on extinction probability. The population exhibited strong survival and reproduction, and the model predicted stable median population growth and a 15% probability of extinction over 50 years in the absence of inbreeding depression. However, our model also predicted the population will lose 40–57% of its heterozygosity in 50 years. When we reduced demographic parameters proportional to reductions documented in another wild population of mountain lions that experienced inbreeding depression, extinction probability rose to 99.7%. Simulating greater landscape connectivity by increasing immigration to greater than or equal to one migrant per generation appears sufficient to largely maintain genetic diversity and reduce extinction probability. We provide empirical support for the central tenet of the extinction vortex as interactions between genetics and demography greatly increased extinction probability relative to the risk from demographic and environmental stochasticity alone. Our modelling approach realistically integrates demographic and genetic data to provide a comprehensive assessment of factors threatening small populations.

Understanding how landscape, host, and pathogen traits contribute to disease exposure requires systematic evaluations of pathogens within and among host species and geographic regions. The relative importance of these attributes is critical for management of wildlife and mitigating domestic animal and human disease, particularly given rapid ecological changes, such as urbanization. We screened > 1000 samples from sympatric populations of puma (Puma concolor), bobcat (Lynx rufus), and domestic cat (Felis catus) across urban gradients in six sites, representing three regions, in North America for exposure to a representative suite of bacterial, protozoal, and viral pathogens (Bartonella sp., Toxoplasma gondii, feline herpesvirus-1, feline panleukopenea virus, feline calicivirus, and feline immunodeficiency virus). We evaluated prevalence within each species, and examined host trait and land cover determinants of exposure; providing an unprecedented analysis of factors relating to potential for infections in domesticated and wild felids. Prevalence differed among host species (highest for puma and lowest for domestic cat) and was greater for indirectly transmitted pathogens. Sex was inconsistently predictive of exposure to directly transmitted pathogens only, and age infrequently predictive of both direct and indirectly transmitted pathogens. Determinants of pathogen exposure were widely divergent between the wild felid species. For puma, suburban land use predicted increased exposure to Bartonella sp. in southern California, and FHV-1 exposure increased near urban edges in Florida. This may suggest interspecific transmission with domestic cats via flea vectors (California) and direct contact (Florida) around urban boundaries. Bobcats captured near urban areas had increased exposure to T. gondii in Florida, suggesting an urban source of prey Bobcats captured near urban areas in Colorado and Florida had higher FIV exposure, possibly suggesting increased intraspecific interactions through pile-up of home ranges. Beyond these regional and pathogen specific relationships, proximity to the wildland-urban interface did not generally increase the probability of disease exposure in wild or domestic felids, empha- sizing the importance of local ecological determinants. Indeed, pathogen exposure was often negatively associated with the wildland-urban interface for all felids. Our analyses suggest cross-species pathogen transmission events around this interface may be infrequent, but followed by self-sustaining propagation within the new host species. virus; puma (Puma concolor); Toxoplasma gondii; urbanization.

Understanding how human activities influence immune response to environmental stressors can support biodiversity conservation across increasingly urbanizing landscapes. We studied a bobcat ( Lynx rufus ) population in urban southern California that experienced a rapid population decline from 2002–2005 due to notoedric mange. Because anticoagulant rodenticide (AR) exposure was an underlying complication in mange deaths, we aimed to understand sublethal contributions of urbanization and ARs on 65 biochemical markers of immune and organ function. Variance in immunological variables was primarily associated with AR exposure and secondarily with urbanization. Use of urban habitat and AR exposure has pervasive, complex and predictable effects on biochemical markers of immune and organ function in free-ranging bobcats that include impacts on neutrophil, lymphocyte and cytokine populations, total bilirubin and phosphorus. We find evidence of both inflammatory response and immune suppression associated with urban land use and rat poison exposure that could influence susceptibility to opportunistic infections. Consequently, AR exposure may influence mortality and has population-level effects, as previous work in the focal population has revealed substantial mortality caused by mange infection. The secondary effects of anticoagulant exposure may be a worldwide, largely unrecognized problem affecting a variety of vertebrate species in human-dominated environments.

Anticoagulant rodenticides (ARs) are increasingly recognized as a threat to nontarget wildlife. High exposure to ARs has been documented globally in nontarget predatory species and linked to the high prevalence of an ectoparasitic disease, notoedric mange. In southern California, mange associated with AR exposure has been the proximate cause of a bobcat (Lynx rufus) population decline. We measured AR exposure in bobcats from two areas in southern California, examining seasonal, demographic and spatial risk factors across landscapes including natural and urbanized areas. The long-term study included bobcats sampled over a 16-year period (1997–2012) and a wide geographic area. We sampled blood (N = 206) and liver (N = 172) to examine exposure ante- and post-mortem. We detected high exposure prevalence (89 %, liver; 39 %, blood) and for individuals with paired liver and blood data (N = 64), 92 % were exposed. Moreover, the animals with the most complete sampling were exposed most frequently to three or more compounds. Toxicant exposure was associated with commercial, residential, and agricultural development. Bobcats of both sexes and age classes were found to be at high risk of exposure, and we documented fetal transfer of multiple ARs. We found a strong association between certain levels of exposure (ppm), and between multiple AR exposure events, and notoedric mange. AR exposure was prevalent throughout both regions sampled and throughout the 16-year time period in the long-term study. ARs pose a substantial threat to bobcats, and likely other mammalian and avian predators, living at the urban-wildland interface.

Mountain lions ( Puma concolor ) throughout North and South America are infected with puma lentivirus clade B (PLVB). A second, highly divergent lentiviral clade, PLVA, infects mountain lions in southern California and Florida. Bobcats ( Lynx rufus ) in these two geographic regions are also infected with PLVA, and to date, this is the only strain of lentivirus identified in bobcats. We sequenced full-length PLV genomes in order to characterize the molecular evolution of PLV in bobcats and mountain lions. Low sequence homology (88% average pairwise identity) and frequent recombination (1 recombination breakpoint per 3 isolates analyzed) were observed in both clades. Viral proteins have markedly different patterns of evolution; sequence homology and negative selection were highest in Gag and Pol and lowest in Vif and Env. A total of 1.7% of sites across the PLV genome evolve under positive selection, indicating that host-imposed selection pressure is an important force shaping PLV evolution. PLVA strains are highly spatially structured, reflecting the population dynamics of their primary host, the bobcat. In contrast, the phylogeography of PLVB reflects the highly mobile mountain lion, with diverse PLVB isolates cocirculating in some areas and genetically related viruses being present in populations separated by thousands of kilometers. We conclude that PLVA and PLVB are two different viral species with distinct feline hosts and evolutionary histories. IMPORTANCE An understanding of viral evolution in natural host populations is a fundamental goal of virology, molecular biology, and disease ecology. Here we provide a detailed analysis of puma lentivirus (PLV) evolution in two natural carnivore hosts, the bobcat and mountain lion. Our results illustrate that PLV evolution is a dynamic process that results from high rates of viral mutation/recombination and host-imposed selection pressure.

Gammaherpesviruses (GHVs) are a diverse and rapidly expanding group of viruses associated with a variety of disease conditions in humans and animals. To identify felid GHVs, we screened domestic cat ( Felis catus ), bobcat ( Lynx rufus ), and puma ( Puma concolor ) blood cell DNA samples from California, Colorado, and Florida using a degenerate pan-GHV PCR. Additional pan-GHV and long-distance PCRs were used to sequence a contiguous 3.4-kb region of each putative virus species, including partial glycoprotein B and DNA polymerase genes. We identified three novel GHVs, each present predominantly in one felid species: Felis catus GHV 1 (FcaGHV1) in domestic cats, Lynx rufus GHV 1 (LruGHV1) in bobcats, and Puma concolor GHV 1 (PcoGHV1) in pumas. To estimate infection prevalence, we developed real-time quantitative PCR assays for each virus and screened additional DNA samples from all three species ( n = 282). FcaGHV1 was detected in 16% of domestic cats across all study sites. LruGHV1 was detected in 47% of bobcats and 13% of pumas across all study sites, suggesting relatively common interspecific transmission. PcoGHV1 was detected in 6% of pumas, all from a specific region of Southern California. The risk of infection for each host varied with geographic location. Age was a positive risk factor for bobcat LruGHV1 infection, and age and being male were risk factors for domestic cat FcaGHV1 infection. Further characterization of these viruses may have significant health implications for domestic cats and may aid studies of free-ranging felid ecology. IMPORTANCE Gammaherpesviruses (GHVs) establish lifelong infection in many animal species and can cause cancer and other diseases in humans and animals. In this study, we identified the DNA sequences of three GHVs present in the blood of domestic cats ( Felis catus ), bobcats ( Lynx rufus ), and pumas ( Puma concolor ; also known as mountain lions, cougars, and panthers). We found that these viruses were closely related to, but distinct from, other known GHVs of animals and represent the first GHVs identified to be native to these feline species. We developed techniques to rapidly and specifically detect the DNA of these viruses in feline blood and found that the domestic cat and bobcat viruses were widespread across the United States. In contrast, puma virus was found only in a specific region of Southern California. Surprisingly, the bobcat virus was also detected in some pumas, suggesting relatively common virus transmission between these species. Adult domestic cats and bobcats were at greater risk for infection than juveniles. Male domestic cats were at greater risk for infection than females. This study identifies three new viruses that are widespread in three feline species, indicates risk factors for infection that may relate to the route of infection, and demonstrates cross-species transmission between bobcats and pumas. These newly identified viruses may have important effects on feline health and ecology.

Urbanization drastically reduces and fragments wildlife habitat, but it can also generate less obvious effects that influence wildlife populations and communities. Two important influences that are often imperceptible without intense scientific investigation are infectious disease and contaminants. In its broadest sense, disease (“dis” and “ease,” or the lack of ease) includes the effects of contaminants, but here we will use the term disease to refer to infectious diseases caused by living organisms, including microparasites, (e.g., bacteria and viruses) and macroparasites (e.g., arthropods and worms). Disease and contaminant effects can be cryptic, unless there are large, conspicuous die-offs. Furthermore, diseased animals are hard to find (unless radio-tracked), and even if they are found, lack of clear symptoms, lack of interest in cause of death, or lack of resources make diagnosis difficult.

Background Since the emergence of next generation sequencing platforms, unprecedented opportunities have arisen in the study of natural vertebrate populations. In particular, insights into the genetic and epigenetic mechanisms of adaptation can be revealed through study of the expression profiles of genes. However, as a pre-requisite to expression profiling, care must be taken in RNA preparation as factors like DNA contamination, RNA integrity or transcript abundance can affect downstream applications. Here, we evaluated five commonly used RNA extraction methods using whole blood sampled under varying conditions from 20 wild carnivores. Results Despite the use of minute starting volumes, all methods produced quantifiable RNA extracts (1.4 – 18.4 μg) with varying integrity (RIN 4.6 - 7.7), the latter being significantly affected by the storage and extraction method used. We observed a significant overall effect of the extraction method on DNA contamination. One particular extraction method, the LeukoLOCK™ filter system, yielded high RNA integrity along with low DNA contamination and efficient depletion of hemoglobin transcripts highly abundant in whole blood. In a proof of concept sequencing experiment, we found globin RNA transcripts to occupy up to ¼ of all sequencing reads if libraries were not depleted of hemoglobin prior to sequencing. Conclusion By carefully choosing the appropriate RNA extraction method, whole blood can become a valuable source for high-throughput applications like expression arrays or transcriptome sequencing from natural populations. Additionally, candidate genes showing signs of selection could subsequently be genotyped in large population samples using whole blood as a source for RNA without harming individuals from rare or endangered species.

Author(s): Moriarty, Joanne Gale; Riley, Seth P. D.; Serieys, Laurel E.; Sikich, Jeff A.; Schoonmaker, Cathy M.; Poppenga, Robert H. | Abstract: A 15-year study of carnivores in an urban landscape in southern California has revealed a high incidence of exposure of non-target wildlife to anticoagulant rodenticides (ARs). All carnivore species studied, including mountain lions, coyotes, bobcats, and gray foxes, have tested positive for exposure to the toxicants. Anticoagulant residues have been detected in post-mortem liver samples at a rate of 83-93% of individuals tested, for coyotes, bobcats, and mountain lions, the three species for which we have extensive sampling. We have also documented mortalities caused directly by exposure to ARs in all four species, particularly in the canids. In both felid species, we found a positive correlation between AR exposure and mange disease, specifically notoedric mange. The incidence of fatal mange infection in bobcats has been at epizootic levels since 2002 in our study area, and more recently outbreaks of the disease have been documented in several other populations in California, all apparently (where testing has been done) in association with exposure to ARs. There are no previously reported instances of epizootics of notoedric mange in any wild felid population. Carnivore exposure to these toxicants appears to be largely secondary (or tertiary, as may be the case in mountain lions) through consumption of their natural prey. In our most recent work, we have evaluated AR exposure of carnivore prey species, including rodents and lagomorphs. We have documented exposure in ground squirrels and woodrats, both of which are regularly consumed by gray foxes, coyotes, and bobcats. Overall, we have found widespread exposure of non-target wildlife to these toxicants, with potentially significant consequences for some species.