Draft Genome Sequences of 158 Listeria monocytogenes Strains Isolated from Black Bears (Ursus americanus) in the United States.

Listeria monocytogenes is responsible for severe foodborne disease and major economic losses, but its potential reservoirs in natural ecosystems remain poorly understood. Here, we report the draft genome sequences of 158 L. monocytogenes strains isolated from black bears (Ursus americanus) in the southeastern United States between 2014 and 2017.

Madagascar Terrestrial Camera Survey Database 2021: A collation of protected forest camera surveys from 2007-2021.

Madagascar is a threatened global biodiversity hotspot and conservation priority, yet we lack broad-scale surveys to assess biodiversity across space and time. To fill this gap, we collated camera trap surveys, capturing species occurrences within Madagascar into a single standardized database. This data set includes nine distinct protected areas of Madagascar and encompasses 13 subprojects, 38 camera arrays, and 1156 sampling units (independent camera site per survey) within two important biodiversity eco-regions: western dry deciduous forest and eastern humid rainforest. Camera surveys were conducted from June 2007 to January 2021. The final data set includes 17 unique families of mammals (Bovidae, Canidae, Cheirogaleidae, Daubentoniidae, Equidae, Eupleridae, Felidae, Hominidae, Indriidae, Lemuridae, Lepilemuridae, Muridae, Nesomyidae, Pteropodidae, Soricidae, Suidae, Tenrecidae) comprising 45 species and 27 unique families of birds (Accipitridae, Acrocephalidae, Alcedinidae, Bernieridae, Brachypteraciidae, Caprimulgidae, Cisticolidae, Columbidae, Coraciidae, Corvidae, Cuculidae, Dicruridae, Mesitornithidae, Monarchidae, Motacillidae, Muscicapidae, Numididae, Phasianidae, Rallidae, Sarothruridae, Strigidae, Sturnidae, Sulidae, Threskiornithidae, Upupidae, Vangidae, Zosteropidae) comprising 58 species. Images were processed and verified by individual project data set creators and camera operation and species tables were then collated. The final product represents the first broad-scale freely available standardized formal faunal database for Madagascar. Data are available through this publication and at DOI: 10.5281/zenodo.5801806. These data will be useful for examining species-level and community-level trends in occurrence across space or time within Madagascar and globally, evaluating native and invasive species dynamics, and will aid in determining species conservation status and planning for at-risk species. There are no copyright restrictions; please cite this paper when using the data for publication.

Assessing carnivore spatial co-occurrence and temporal overlap in the face of human interference in a semiarid forest.

Apex predators drive top-down effects in ecosystems and the loss of such species can trigger mesopredator release. This ecological process has been well documented in human-modified small areas, but for management and conservation of ecological communities, it is important to know which human factors affect apex predator occurrence and which mediate mesopredators release at large scales. We hypothesized that mesopredators would avoid spatial and temporal overlap with the apex predator, the puma; but that human perturbations (i.e., cattle raising and trophy hunting) would dampen top-down effects and mediate habitat use. We installed 16 camera traps in each of 45, 10 × 10 km grid cells in the Caldén forest region of central Argentina resulting in 706 total stations covering 61,611 km2 . We used single-season occupancy and two-species co-occurrence models and calculated the species interaction factor (SIF) to explore the contributions of habitat, biotic, and anthropic variables in explaining co-occurrence between carnivore pairs. We also used kernel density estimation techniques to analyze temporal overlap in activity patterns of the carnivore guild. We found that puma habitat use increased with abundance of large prey and with proximity to protected areas. Geoffroy's cats and skunks spatially avoided pumas and this effect was strong and mediated by distance to protected areas and game reserves, but pumas did not influence pampas fox and pampas cat space use. At medium and low levels of puma occupancy, we found evidence of spatial avoidance between three pairs of mesocarnivores. All predators were mostly nocturnal and crepuscular across seasons and mesopredators showed little consistent evidence of changing activity patterns with varying levels of puma occupancy or human interference. We found potential for mesopredator release at large scale, especially on the spatial niche axis. Our results suggest that a combination of interacting factors, in conjunction with habitat features and intervening human activities, may make mesopredator release unlikely or difficult to discern at broad scales. Overall, we believe that promoting the creation of new protected areas linked by small forest patches would likely lead to increased predator and prey abundances, as well as the interactions among carnivores inside and outside of protected areas.

Sex-specific effects of reproductive season on bobcat space use, movement, and resource selection in the Appalachian Mountains of Virginia.

Across taxa, sex-specific demands vary temporally in accordance with reproductive investments. In solitary carnivores, females must provision and protect young independently while meeting increased energetic demands. Males seek to monopolize access to females by maintaining large territories and defending them from other males. For many species, it is poorly understood how these demands relate to broad-scale animal movements. To investigate predictions surrounding the reproductive strategies of solitary carnivores and effects of local conditions on bobcat (Lynx rufus) spatial ecology, we examined the effects of sex and reproductive season on home range size, movement rate, and resource selection of bobcats in the central Appalachian Mountains. Male seasonal home ranges were approximately 3 times larger than those of females (33.9 ± 2.6 vs. 12.1 ± 2.4 km2, x±SE), and male movement rates were 1.4 times greater than females (212.6 ± 3.6 vs. 155 ± 8.2 m/hr), likely reflecting male efforts to maximize access to females. Both sexes appear to maintain relatively stable seasonal home ranges despite temporally varying reproductive investments, instead adjusting movements within home ranges. Males increased movements during the dispersal period, potentially reflecting increased territoriality prior to breeding. Females increased movements during the kitten-rearing period, when foraging more intensively, and frequently returning to den sites. Both sexes selected home ranges at higher elevations. However, females selected deciduous forest and avoided fields, whereas males selected fields and avoided deciduous forest, perhaps explained by male pressure to access multiple females across several mountain ridges and higher risk tolerance. Seasonal changes in home range selection likely reflect changes in home range shape. Increased female avoidance of fields during kitten rearing may indicate female avoidance of presumably resource rich, yet risky, fields at the time when kittens are most vulnerable. Our results indicate that while reproductive chronology influences the spatial ecology of solitary carnivores, effects may be constrained by territoriality.

Listeria monocytogenes at the human-wildlife interface: black bears (Ursus americanus) as potential vehicles for Listeria.

Listeria monocytogenes is the causative agent of the foodborne illness listeriosis, which can result in severe symptoms and death in susceptible humans and other animals. L. monocytogenes is ubiquitous in the environment and isolates from food and food processing, and clinical sources have been extensively characterized. However, limited information is available on L. monocytogenes from wildlife, especially from urban or suburban settings. As urban and suburban areas are expanding worldwide, humans are increasingly encroaching into wildlife habitats, enhancing the frequency of human-wildlife contacts and associated pathogen transfer events. We investigated the prevalence and characteristics of L. monocytogenes in 231 wild black bear capture events between 2014 and 2017 in urban and suburban sites in North Carolina, Georgia, Virginia and United States, with samples derived from 183 different bears. Of the 231 captures, 105 (45%) yielded L. monocytogenes either alone or together with other Listeria. Analysis of 501 samples, primarily faeces, rectal and nasal swabs for Listeria spp., yielded 777 isolates, of which 537 (70%) were L. monocytogenes. Most L. monocytogenes isolates exhibited serotypes commonly associated with human disease: serotype 1/2a or 3a (57%), followed by the serotype 4b complex (33%). Interestingly, approximately 50% of the serotype 4b isolates had the IVb-v1 profile, associated with emerging clones of L. monocytogenes. Thus, black bears may serve as novel vehicles for L. monocytogenes, including potentially emerging clones. Our results have significant public health implications as they suggest that the ursine host may preferentially select for L. monocytogenes of clinically relevant lineages over the diverse listerial populations in the environment. These findings also help to elucidate the ecology of L. monocytogenes and highlight the public health significance of the human-wildlife interface.

Gut microbiota and their putative metabolic functions in fragmented Bengal tiger population of Nepal.

Bengal tigers (Panthera tigris tigris) serve a pivotal role as an apex predator in forest ecosystems. To increase our knowledge on factors impacting the viability and health of this endangered species, we studied the gut microbiota in 32 individual Bengal tigers from three geographically separated areas (Chitwan National Park (CNP), Bardia National Park (BNP) and Suklaphanta Wildlife Reserve (SWR)) in Nepal, using noninvasive genetic sampling methods. Gut microbiota influence the immune system, impact various physiological functions, and modulates metabolic reactions, that ultimately impact the host health, behavior and development. Across the tiger populations in Nepal, we found significant differences in the composition of microbial communities based on their geographic locations. Specifically, we detected significant differences between CNP and the other two protected areas (CNP vs BNP: pseudo t = 1.944, P = 0.006; CNP vs SWR: pseudo t = 1.9942, P = 0.0071), but no differences between BNP and SWR. This mirrors what has been found for tiger gene flow in the same populations, suggesting gut microbiota composition and host gene flow may be linked. Furthermore, predictive metagenome functional content analysis (PICRUSt) revealed a higher functional enrichment and diversity for significant gut microbiota in the Chitwan tiger population and the lowest enrichment and diversity in Suklaphanta. The CNP tiger population contained higher proportions of microbiota that are associated with predicted functions relevant for metabolism of amino acid, lipid, xenobiotics biodegradation, terpenoides and polyketides than the SWR population. We conclude the tiger population structure, gut microbiota profile and associated functional metabolic categories are correlated, with geographically most separated CNP and SWR tiger population having the most distinct and different host genotype and microbiota profiles. Our work dramatically expands the understanding of tiger microbiota in wild populations and provides a valuable case study on how to investigate genetic diversity at different hierarchical levels, including hosts as well as their microbial communities.

Does faecal matter reflect location? An initial assessment of isotopic variability between consumed prey remains and faecal matter for wild jaguars.

Faecal isotopic analysis may complement other non-invasive wildlife survey tools for monitoring landscape use by carnivores, such as motion-detecting cameras and non-invasive genetic sampling. We analysed carbon, nitrogen, and strontium isotopes in faecal matter produced by jaguars (Panthera onca) as well as bones from consumed prey at the Mountain Pine Ridge Forest Reserve (MPR) in Belize, Central America. The MPR is ideally suited for a spatial isotope study as vegetation and geology both vary considerably. The isotopic composition of faecal matter should reflect the habitat and geology where consumed prey lived. We used bone from consumed prey recovered from jaguar scats as a proxy for diet. Faecal matter and bone showed comparable spatial isotopic trends, suggesting that the isotopic composition of jaguar faeces can be used to detect foraging in different habitats (pine forest versus broadleaf forest) or on different geologies (Mesozoic carbonates; Palaeozoic granite, contact metamorphics, and metasediments). This result is reassuring as bones are not always present in carnivore scats. Studying landscape use by cryptic and wide-ranging carnivore species like jaguars remains challenging. Isotopic analysis of faecal matter complements the existing array of non-invasive spatial monitoring tools.

The road to deforestation: Edge effects in an endemic ecosystem in Sumatra, Indonesia.

Worldwide, roads are a main driver of deforestation and degradation as they increase forest access along the forest edge. In many tropical areas, unofficial roads go unreported and unrecorded, resulting in inaccurate estimates of intact forested areas. This is the case in central Sumatra, which boasts populations of critically endangered Sumatran elephants (Elephas maximus sumatrensis), tigers (Panthera tigris sumatrae) and other endemic flora and fauna that make the area globally unique. However, maps do not reflect the reality of forest loss in the area. Here we present new maps from 2002 and 2016 of digitized and ground-truthed roads in one of Sumatra's unique lowland tropical protected areas, Tesso Nilo National Park. Using our newly created roads dataset, we examine the distribution of forest with respect to distance to roads. Our data show >2,400 km of roads within the national park in 2016 -nearly a 10-fold increase from roads known in 2002. Most forest (82-99%) within Tesso Nilo falls within 100 m, 500 m, and 1000 m of road edges. Length of road increased 157% and road density increased from 1.06 km/km2 to 2.63 km/km2 from 2002-2016. Our results suggest that this endemic ecosystem is facing substantial threat from roads and their associated impacts. Without swift management action, such as road closures and increased enforcements by park management, this ecosystem, and its endemic wildlife, could be lost. It is imperative that protected areas worldwide more rigorously consider roads and road effects on ecosystem fragmentation in their conservation plans.

Elephant (Elephas maximus) temporal activity, distribution, and habitat use patterns on the tiger's forgotten trails across the seasonally dry, subtropical, hilly Churia forests of Nepal.

Understanding spatial distribution, habitat use, and temporal activity patterns is important for species conservation planning. This information especially is crucial for mega herbivores like elephants as their ranging patterns encompass a myriad of habitats types. Churia habitat is geological fragile yet important for wildlife in Nepal and India. We used camera trapping and sign surveys covering 536 km2 of Churia and surrounding areas within Chitwan National Park. Across 152 trapping locations, we accumulated 2,097 trap nights in a 60-day survey during the winter season of 2010-11. We used a non-parametric kernel density function to analyze winter activity patterns of elephants detected in camera-traps. Additionally, we walked 643 km over 76 grid cells in two surveys (winter and summer) to estimate elephant distribution and intensity of habitat use using an occupancy framework. Multi-season models allowed us to make seasonal (winter versus summer) inferences regarding changes in habitat use based on covariates influencing use and detection. We photographed 25 mammalian species including elephants with calves with a trapping rate of 2.72 elephant photos events per 100 trap nights. Elephant winter activity pattern was found to be mainly nocturnal, with crepuscular peaks. Covariates such as normalized differential vegetation index and terrain ruggedness positively influenced elephant spatial distribution and habitat use patterns within the Churia habitat. We also found lower elephant habitat use ([Formula: see text]) of Churia in winter 0.51 (0.02) than in summer 0.57 (0.02). Elephants heavily used the eastern portion of Churia in both seasons (67-69%). Overall, Churia habitat, which is often ignored, clearly is used by elephants, with increases in summer use in the west and high use year-round in the east, and thus should no longer be neglected or forgotten in species conservation planning.

Mapping and predicting forest loss in a Sumatran tiger landscape from 2002 to 2050.

Riau Province in central Sumatra, with its peatland, lowland, and montane forest habitats, was once a stronghold for Sumatran tiger (Panthera tigris sumatrae) populations. Today, Riau may have one of the highest deforestation rates in the world and wildlife populations are dwindling, with natural forest now comprising approximately only 18% of the province, mostly contained within protected areas. Agriculture (acacia, rubber, and oil palm) makes up the majority of Riau's land cover and deforestation for the creation of new plantations is rampant. Natural forest and tigers still remain in Bukit Tigapuluh National Park and Rimbang Baling Wildlife Reserve, which remain connected to tiger populations in montane forest on the western edge of Sumatra. In this study, using freely available Landsat imagery and a maximum likelihood classification algorithm, we create land cover maps for central Sumatra from 2002 to 2016. We then use current land cover, elevation, and slope variables to predict changes from forest to plantation from 2016 to 2050 at five year intervals using a multilayer perceptron neural network. Finally, we compare connectivity based on a 100 km distance threshold (based on potential tiger dispersal) across the landscape and across years. Land cover maps had 80-90% accuracy, and we predict forest in Tesso Nilo and the western edge of the study area to be lost by 2050 given current rates of deforestation. Our connectivity analysis shows that Tesso Nilo and the area between Rimbang Baling and Bukit Tigapuluh are important components for maintaining connectivity throughout the study area. Focusing conservation and rehabilitation efforts on forests close to plantations in flat areas, including Tesso Nilo, is necessary to maintain forests and increase connectivity in Riau to ensure future habitat connectivity for survival of tigers and Sumatra's other diverse endemic species.

Species, sex and geo-location identification of seized tiger (Panthera tigris tigris) parts in Nepal-A molecular forensic approach.

Tiger (Panthera tigris) populations are in danger across their entire range due to habitat loss, poaching and the demand for tiger parts. The Bengal tiger (Panthera tigris tigris) is an endangered apex predator with a population size estimated to be less than 200 in Nepal. In spite of strict wildlife protection laws, illegal trade of tiger parts is increasing; and Nepal has become one of the major sources and transit routes for poached wildlife parts. Identification of wildlife parts is often challenging for law enforcement officials due to inadequate training and lack of available tools. Here, we describe a molecular forensic approach to gain insight into illegally trafficked tiger parts seized across Nepal. We created Nepal's first comprehensive reference genetic database of wild tigers through the Nepal Tiger Genome Project (2011-2013). This database has nuclear DNA microsatellite genotype and sex profiles, including geo-spatial information, of over 60% (n = 120) of the wild tigers of Nepal. We analyzed 15 putative cases of confiscated poached tiger parts and all were confirmed to be of tiger. Ten samples were identified as male and five were female. We determined probable geo-source location for 9 of the 14 samples with 6-8 nuclear DNA microsatellite loci using inferences from four different statistical assignment methods. Six samples were assigned to Bardia National Park and one of these was an exact match to a female tiger previously profiled in our fecal DNA reference database. Two tiger samples were assigned to Shuklaphanta Wildlife Reserve and one to Chitwan National Park. We are unable to definitively assign five tiger samples which could be offspring dispersers or might have come from tiger population outside of Nepal. Our study revealed that the western region, particularly Bardia National Park, is a poaching hotspot for illegal tiger trade in Nepal. We present feasibility of using molecular forensic based evidence to incriminate criminals in a court of law in the fight against wildlife crime.

Ecological correlates of the spatial co-occurrence of sympatric mammalian carnivores worldwide.

The composition of local mammalian carnivore communities has far-reaching effects on terrestrial ecosystems worldwide. To better understand how carnivore communities are structured, we analysed camera trap data for 108 087 trap days across 12 countries spanning five continents. We estimate local probabilities of co-occurrence among 768 species pairs from the order Carnivora and evaluate how shared ecological traits correlate with probabilities of co-occurrence. Within individual study areas, species pairs co-occurred more frequently than expected at random. Co-occurrence probabilities were greatest for species pairs that shared ecological traits including similar body size, temporal activity pattern and diet. However, co-occurrence decreased as compared to other species pairs when the pair included a large-bodied carnivore. Our results suggest that a combination of shared traits and top-down regulation by large carnivores shape local carnivore communities globally.

Assessment of genetic diversity, population structure, and gene flow of tigers (Panthera tigris tigris) across Nepal's Terai Arc Landscape.

With fewer than 200 tigers (Panthera tigris tigris) left in Nepal, that are generally confined to five protected areas across the Terai Arc Landscape, genetic studies are needed to provide crucial information on diversity and connectivity for devising an effective country-wide tiger conservation strategy. As part of the Nepal Tiger Genome Project, we studied landscape change, genetic variation, population structure, and gene flow of tigers across the Terai Arc Landscape by conducting Nepal's first comprehensive and systematic scat-based, non-invasive genetic survey. Of the 770 scat samples collected opportunistically from five protected areas and six presumed corridors, 412 were tiger (57%). Out of ten microsatellite loci, we retain eight markers that were used in identifying 78 individual tigers. We used this dataset to examine population structure, genetic variation, contemporary gene flow, and potential population bottlenecks of tigers in Nepal. We detected three genetic clusters consistent with three demographic sub-populations and found moderate levels of genetic variation (He = 0.61, AR = 3.51) and genetic differentiation (FST = 0.14) across the landscape. We detected 3-7 migrants, confirming the potential for dispersal-mediated gene flow across the landscape. We found evidence of a bottleneck signature likely caused by large-scale land-use change documented in the last two centuries in the Terai forest. Securing tiger habitat including functional forest corridors is essential to enhance gene flow across the landscape and ensure long-term tiger survival. This requires cooperation among multiple stakeholders and careful conservation planning to prevent detrimental effects of anthropogenic activities on tigers.

Long term monitoring of jaguars in the Cockscomb Basin Wildlife Sanctuary, Belize; Implications for camera trap studies of carnivores.

In this study, we estimate life history parameters and abundance for a protected jaguar population using camera-trap data from a 14-year monitoring program (2002-2015) in Belize, Central America. We investigated the dynamics of this jaguar population using 3,075 detection events of 105 individual adult jaguars. Using robust design open population models, we estimated apparent survival and temporary emigration and investigated individual heterogeneity in detection rates across years. Survival probability was high and constant among the years for both sexes (φ = 0.78), and the maximum (conservative) age recorded was 14 years. Temporary emigration rate for the population was random, but constant through time at 0.20 per year. Detection probability varied between sexes, and among years and individuals. Heterogeneity in detection took the form of a dichotomy for males: those with consistently high detection rates, and those with low, sporadic detection rates, suggesting a relatively stable population of 'residents' consistently present and a fluctuating layer of 'transients'. Female detection was always low and sporadic. On average, twice as many males than females were detected per survey, and individual detection rates were significantly higher for males. We attribute sex-based differences in detection to biases resulting from social variation in trail-walking behaviour. The number of individual females detected increased when the survey period was extended from 3 months to a full year. Due to the low detection rates of females and the variable 'transient' male subpopulation, annual abundance estimates based on 3-month surveys had low precision. To estimate survival and monitor population changes in elusive, wide-ranging, low-density species, we recommend repeated surveys over multiple years; and suggest that continuous monitoring over multiple years yields even further insight into population dynamics of elusive predator populations.

Density and carrying capacity in the forgotten tigerland: Tigers in the understudied Nepalese Churia.

While there are numerous wildlife ecology studies in lowland areas of Nepal, there are no in-depth studies of the hilly Churia habitat even though it comprises 7642 km2 of potential wildlife habitat across the Terai Arc. We investigated tiger, leopard and prey densities across this understudied habitat. Our camera trapping survey covered 536 km2 of Churia and surrounding areas within Chitwan National Park (CNP). We used 161 trapping locations and accumulated 2097 trap-nights in a 60-day survey period during the winter season of 2010-2011. In addition, we walked 136 km over 81 different line transects using distance sampling to estimate prey density. We photographed 31 individual tigers, 28 individual leopards and 25 other mammalian species. Spatial capture-recapture methods resulted in lower density estimates for tigers, ranging from 2.3 to 2.9 tigers per 100 km2 , than for leopards, which ranged from 3.3 to 5.1 leopards per 100 km2 . In addition, leopard densities were higher in the core of the Churia compared to surrounding areas. We estimated 62.7 prey animals per 100 km2 with forest ungulate prey (sambar, chital, barking deer and wild pig), accounting for 47% of the total. Based on prey availability, Churia habitat within CNP could potentially support 5.86 tigers per 100 km2 but our density estimates were lower, perhaps indicating that the tiger population is below carrying capacity. Our results demonstrate that Churia habitat should not be ignored in conservation initiatives, but rather management efforts should focus on reducing human disturbance to support higher predator numbers.

Space Use and Habitat Selection by Resident and Transient Red Wolves (Canis rufus).

Recovery of large carnivores remains a challenge because complex spatial dynamics that facilitate population persistence are poorly understood. In particular, recovery of the critically endangered red wolf (Canis rufus) has been challenging because of its vulnerability to extinction via human-caused mortality and hybridization with coyotes (Canis latrans). Therefore, understanding red wolf space use and habitat selection is important to assist recovery because key aspects of wolf ecology such as interspecific competition, foraging, and habitat selection are well-known to influence population dynamics and persistence. During 2009-2011, we used global positioning system (GPS) radio-telemetry to quantify space use and 3rd-order habitat selection for resident and transient red wolves on the Albemarle Peninsula of eastern North Carolina. The Albemarle Peninsula was a predominantly agricultural landscape in which red wolves maintained spatially stable home ranges that varied between 25 km2 and 190 km2. Conversely, transient red wolves did not maintain home ranges and traversed areas between 122 km2 and 681 km2. Space use by transient red wolves was not spatially stable and exhibited shifting patterns until residency was achieved by individual wolves. Habitat selection was similar between resident and transient red wolves in which agricultural habitats were selected over forested habitats. However, transients showed stronger selection for edges and roads than resident red wolves. Behaviors of transient wolves are rarely reported in studies of space use and habitat selection because of technological limitations to observed extensive space use and because they do not contribute reproductively to populations. Transients in our study comprised displaced red wolves and younger dispersers that competed for limited space and mating opportunities. Therefore, our results suggest that transiency is likely an important life-history strategy for red wolves that facilitates metapopulation dynamics through short- and long-distance movements and eventual replacement of breeding residents lost to mortality.

A Comparative Analysis of Genetic Diversity and Structure in Jaguars (Panthera onca), Pumas (Puma concolor), and Ocelots (Leopardus pardalis) in Fragmented Landscapes of a Critical Mesoamerican Linkage Zone.

With increasing anthropogenic impact and landscape change, terrestrial carnivore populations are becoming more fragmented. Thus, it is crucial to genetically monitor wild carnivores and quantify changes in genetic diversity and gene flow in response to these threats. This study combined the use of scat detector dogs and molecular scatology to conduct the first genetic study on wild populations of multiple Neotropical felids coexisting across a fragmented landscape in Belize, Central America. We analyzed data from 14 polymorphic microsatellite loci in 1053 scat samples collected from wild jaguars (Panthera onca), pumas (Puma concolor), and ocelots (Leopardus pardalis). We assessed levels of genetic diversity, defined potential genetic clusters, and examined gene flow for the three target species on a countrywide scale using a combination of individual- and population-based analyses. Wild felids in Belize showed moderate levels of genetic variation, with jaguars having the lowest diversity estimates (HE = 0.57 ± 0.02; AR = 3.36 ± 0.09), followed by pumas (HE = 0.57 ± 0.08; AR = 4.20 ± 0.16), and ocelots (HE = 0.63 ± 0.03; AR = 4.16 ± 0.08). We observed low to moderate levels of genetic differentiation for all three target species, with jaguars showing the lowest degree of genetic subdivision across the country, followed by ocelots and pumas. Although levels of genetic diversity and gene flow were still fairly high, we detected evidence of fine-scale genetic subdivision, indicating that levels of genetic connectivity for wild felids in Belize are likely to decrease if habitat loss and fragmentation continue at the current rate. Our study demonstrates the value of understanding fine-scale patterns of gene flow in multiple co-occurring felid species of conservation concern, which is vital for wildlife movement corridor planning and prioritizing future conservation and management efforts within human-impacted landscapes.

Hunting, Exotic Carnivores, and Habitat Loss: Anthropogenic Effects on a Native Carnivore Community, Madagascar.

The wide-ranging, cumulative, negative effects of anthropogenic disturbance, including habitat degradation, exotic species, and hunting, on native wildlife has been well documented across a range of habitats worldwide with carnivores potentially being the most vulnerable due to their more extinction prone characteristics. Investigating the effects of anthropogenic pressures on sympatric carnivores is needed to improve our ability to develop targeted, effective management plans for carnivore conservation worldwide. Utilizing photographic, line-transect, and habitat sampling, as well as landscape analyses and village-based bushmeat hunting surveys, we provide the first investigation of how multiple forms of habitat degradation (fragmentation, exotic carnivores, human encroachment, and hunting) affect carnivore occupancy across Madagascar's largest protected area: the Masoala-Makira landscape. We found that as degradation increased, native carnivore occupancy and encounter rates decreased while exotic carnivore occupancy and encounter rates increased. Feral cats (Felis species) and domestic dogs (Canis familiaris) had higher occupancy than half of the native carnivore species across Madagascar's largest protected landscape. Bird and small mammal encounter rates were negatively associated with exotic carnivore occupancy, but positively associated with the occupancy of four native carnivore species. Spotted fanaloka (Fossa fossana) occupancy was constrained by the presence of exotic feral cats and exotic small Indian civet (Viverricula indica). Hunting was intense across the four study sites where hunting was studied, with the highest rates for the small Indian civet (mean=90 individuals consumed/year), the ring-tailed vontsira (Galidia elegans) (mean=58 consumed/year), and the fosa (Cryptoprocta ferox) (mean=31 consumed/year). Our modeling results suggest hunters target intact forest where carnivore occupancy, abundance, and species richness, are highest. These various anthropogenic pressures and their effects on carnivore populations, especially increases in exotic carnivores and hunting, have wide-ranging, global implications and demand effective management plans to target the influx of exotic carnivores and unsustainable hunting that is affecting carnivore populations across Madagascar and worldwide.

Noninvasive individual and species identification of jaguars (Panthera onca), pumas (Puma concolor) and ocelots (Leopardus pardalis) in Belize, Central America using cross-species microsatellites and faecal DNA.

There is a great need to develop efficient, noninvasive genetic sampling methods to study wild populations of multiple, co-occurring, threatened felids. This is especially important for molecular scatology studies occurring in challenging tropical environments where DNA degrades quickly and the quality of faecal samples varies greatly. We optimized 14 polymorphic microsatellite loci for jaguars (Panthera onca), pumas (Puma concolor) and ocelots (Leopardus pardalis) and assessed their utility for cross-species amplification. Additionally, we tested their reliability for species and individual identification using DNA from faeces of wild felids detected by a scat detector dog across Belize in Central America. All microsatellite loci were successfully amplified in the three target species, were polymorphic with average expected heterozygosities of HE = 0.60 ± 0.18 (SD) for jaguars, HE = 0.65 ± 0.21 (SD) for pumas and HE = 0.70 ± 0.13 (SD) for ocelots and had an overall PCR amplification success of 61%. We used this nuclear DNA primer set to successfully identify species and individuals from 49% of 1053 field-collected scat samples. This set of optimized microsatellite multiplexes represents a powerful tool for future efforts to conduct noninvasive studies on multiple, wild Neotropical felids.

Effects of natural environmental conditions on faecal glucocorticoid metabolite concentrations in†jaguars (Panthera onca) in Belize.

In situ studies that rely on non-invasive faecal hormone monitoring are subject to problems due to potential changes in hormone concentrations in samples exposed to field conditions. In this study, we conducted an environmental validation for measurement of faecal glucocorticoid metabolites (FGMs) in jaguars (Panthera onca). We collected fresh faeces (e.g. no older than 8 h) from jaguars (six males and four females), housed at the Belize Zoo, and exposed them randomly to two environmental conditions: shade and sun. A control (first sub-sample) was immediately frozen, after which sub-samples were frozen daily over a 5 day period in both the dry and wet seasons. We quantified FGMs using a cortisol enzyme immunoassay (EIA) and a corticosterone radioimmunoassay (RIA), both capable of identifying relevant metabolites. Results indicated that FGMs assessed with the cortisol EIA were stable for 5 days during the dry season but for <1 day during the wet season, while FGMs assessed with the corticosterone RIA were stable for 5 days during both the dry and wet seasons. Exposure of jaguar faeces to sun or shade had no effect on FGM concentrations, despite significant differences in weather parameters. Analysis of faecal morphology proved unreliable in identifying faecal age. We conclude that the corticosterone RIA is suitable for assessing FGMs in free-ranging Belizean jaguars by surveying the same transects every 3-4 days in both seasons. The cortisol EIA can be used during the dry season, but there are possible shifts in metabolite immunoactivity in wet conditions. Assessment of adrenal activity in jaguars ranging areas of varying human disturbance is a timely application of this methodology in Belize.