Mortalities in Black Vultures (Coragyps atratus) caused by A/goose/Guangdong/1/1996 2.3.4.4b lineage highly pathogenic (HP) influenza A virus (IAV) H5N1 (HP H5N1) were detected in Florida, USA, on 14 February 2022. By the end of 2022, at least 2,674 and 5,707 deaths where HP H5N1 infection was confirmed or suspected had been documented in Florida and the USA as a whole, respectively. Reported vulture mortalities at die-off locations nationwide ranged from 1 to 700. In Florida and Pennsylvania, USA, antibodies to both H5 and N1 subtypes of IAV were detected in apparently healthy Black Vultures sampled ≤12 mo after mortality events. Antibodies to these subtypes were not detected in Black Vultures at sites in Kentucky and Tennessee, USA, where HP H5N1 was not detected, nor in Pennsylvania vultures sampled before HP H5N1 introduction into North America. Infections in vultures probably originated through scavenging of infected bird carcasses, but once in the vulture population, HP H5N1 infections may have been maintained by conspecific scavenging. Black Vultures can serve as an indicator species for HP H5N1 in North America, and they may sustain an outbreak after infection rates have declined in other species.

The objective of this study was to characterize Notoedres from Florida, USA and to determine if rabbits and squirrels share the same Notoedres species. From July 2022-November 2024, 19 suspected mange (i.e., alopecia and crusty lesions) cases were identified in 16 eastern cottontail rabbits (Sylvilagus floridanus), an eastern gray squirrel (Sciurus carolinensis), a marsh rabbit (Sylvilagus palustris), and a northern raccoon (Procyon lotor) in Florida. Most were in poor nutritional condition with multifocal alopecia and hyperkeratosis. Histologically, mites were noted in the variably hyperkeratotic stratum corneum with associated serocellular crusts and epidermal hyperplasia. Mites from rabbits and the squirrel were identified as Notoedres centrifera and the raccoon was infested with Notoedres cati. ITS-2 sequences from the squirrel and 11 cottontail rabbits were identical and 100 % similar to a N. centrifera sequence from a Florida cottontail rabbit. The other six N. centrifera sequences from the marsh rabbit and five cottontail rabbits matched those from a Florida marsh rabbit and a Kansas fox squirrel (Sciurus niger). The N. cati ITS-2 sequence was 100 % similar to N. cati. The N. centrifera COI sequences were more variable and fell into two groups that were 79.5 % and 79.8 % similar to S. scabiei. The raccoon N. cati COI sequence was 85.1 % (582/684) and 84.1 % (575/684) similar to Group 1 and Group 2 of N. centrifera, respectively, and 80.3 % similar to S. scabiei. Overall, these data may indicate that notoedric mange is endemic in rabbits and squirrels in Florida, confirm raccoons as hosts for N. cati, and highlight the utility of molecular characterization in mite speciation. Additionally, new sequences of Notoedres were generated that will facilitate future phylogenetic analyses. The biodiversity and ecosystem impacts of notoedric mange in Florida are currently unknown, but this research indicates a need for further field-based surveys to assess these potential risks.

Previously, a high prevalence of piroplasms has been reported from Florida pumas (Puma concolor coryi) from southern Florida. In the current study, we provide biological, morphological, serological, and molecular data on a novel Babesia species. Ring-stage trophozoites were morphologically similar to trophozoites of numerous small babesids of felids including B. leo, B. felis, and Cytauxzoon felis. Parasitemias in Florida pumas were very low (<1%) and hematologic values of 25 Babesia-infected Florida pumas were within normal ranges for P. concolor. Phylogenetic analysis of near full-length 18S rRNA gene, β-tubulin, cytochrome c oxidase subunit I, cytochrome c oxidase subunit III, and cytochrome b gene sequences indicated that this Babesia species is a member of the Babesia sensu stricto clade and is related to groups of Babesia spp. from carnivores or ungulates, although the closest group varied by gene target. Internal transcribed spacer (ITS)-1 region sequences from this Babesia sp. from 19 Florida pumas were 85.7-99.5% similar to each other and ∼88% similar to B. odocoilei. Similarly, an ITS-2 sequence from one puma was 96% similar to B. bigemina and 92% similar to a Babesia sp. from a red panda (Ailurus fulgens). Infected pumas were positive for antibodies that reacted with B. odocoilei, B. canis, and B. bovis antigens with titers of 1:256, 1:128, and 1:128, respectively. No serologic reactivity was noted for Theileria equi. No molecular evidence of congenital infection was detected in 24 kittens born to 11 Babesia-infected female pumas. Pumas from other populations in the United States [Louisiana (n=1), North Dakota (n=5) and Texas (n=28)], British Columbia, Canada (n=9), and Costa Rica (n=2) were negative for this Babesia sp. Collectively, these data indicate that this Babesia sp. is novel and is the first description of a felid-associated Babesia species in North America.

Genetic rescue—an increase in population fitness following the introduction of new alleles—has been proven to ameliorate inbreeding depression in small, isolated populations, yet is rarely applied as a conservation tool. A lingering question regarding genetic rescue in wildlife conservation is how long beneficial effects persist in admixed populations. Using data collected over 40 years from 1192 endangered Florida panthers (Puma concolor coryi) across nine generations, we show that the experimental genetic rescue implemented in 1995—via the release of eight female pumas from Texas—alleviated morphological, genetic, and demographic correlates of inbreeding depression, subsequently preventing extirpation of the population. We present unequivocal evidence, for the first time in any terrestrial vertebrate, that genetic and phenotypic benefits of genetic rescue remain in this population after five generations of admixture, which helped increase panther abundance (> fivefold) and genetic effective population size (> 20-fold). Additionally, even with extensive admixture, microsatellite allele frequencies in the population continue to support the distinctness of Florida panthers from other North American puma populations, including Texas. Although threats including habitat loss, human-wildlife conflict, and infectious diseases are challenges to many imperiled populations, our results suggest genetic rescue can serve as an effective, multi-generational tool for conservation of small, isolated populations facing extinction from inbreeding.

Baylisascaris procyonis, or raccoon roundworm, is an intestinal nematode parasite of raccoons (Procyon lotor) that is important to public and wildlife health. Historically, the parasite was uncommon in the southeastern US; however, the range of B. procyonis has expanded to include Florida, US. From 2010 to 2016, we opportunistically sampled 1,030 raccoons statewide. The overall prevalence was 3.7% (95% confidence interval=2.5-4.8%) of sampled individuals, and infection intensity ranged from 1 to 48 (mean±standard deviation 9.9±4.0). We found raccoon roundworm in 9/56 (16%) counties sampled, and the percent positive ranged from 1.1% to 13.3% of specimens collected per county. Including previously published data, B. procyonis was detected in 11 Florida counties. We used logistic regression to estimate the contribution of raccoon demographic variables and the presence of the endoparasite Macracanthorhynchus ingens to B. procyonis detection in Florida. Following the model selection process we found housing density, M. ingens presence, and urbanicity to be predictive of raccoon roundworm presence. We also found substantial among-county variation. Raccoon sex and age were not useful predictors. Public health officials, wildlife rehabilitators, wildlife managers, and others should consider any Florida raccoon to be potentially infected with B. procyonis, particularly in areas where housing density is high.

Seasonal variation in habitat use and animal behavior can alter host contact patterns with potential consequences for pathogen transmission dynamics. The endangered Florida panther (Puma concolor coryi) has experienced significant pathogen-induced mortality and continues to be at risk of future epidemics. Prior research has found increased panther movement in Florida’s dry versus wet seasons, which may affect panther population connectivity and seasonally increase potential pathogen transmission. Our objective was to determine if Florida panthers are more spatially connected in dry seasons relative to wet seasons, and test if identified connectivity differences resulted in divergent predicted epidemic dynamics. We leveraged extensive panther telemetry data to construct seasonal panther home range overlap networks over an 11 year period. We tested for differences in network connectivity, and used observed network characteristics to simulate transmission of a broad range of pathogens through dry and wet season networks. We found that panthers were more spatially connected in dry seasons than wet seasons. Further, these differences resulted in a trend toward larger and longer pathogen outbreaks when epidemics were initiated in the dry season. Our results demonstrate that seasonal variation in behavioral patterns—even among largely solitary species—can have substantial impacts on epidemic dynamics.

The bald eagle (Haliaeetus leucocephalus) is a culturally and ecologically vital species in North America that embodies conservation success but continues to face threats that include emerging pathogens. The introduction of A/goose/Guangdong/1/1996 lineage highly pathogenic (HP) clade 2.3.4.4b H5N1 influenza A virus (IAV) in North America in late 2021 resulted in high rates of mortality among bald eagles. Here we show an alarming rate of bald eagle nest failure and mortality attributed to HP IAV. We documented fatal, systemic HP IAV infection in breeding adult and nestling bald eagles along the southeastern U.S. coast. Concurrently, annual bald eagle nest surveys in Georgia and Florida revealed a precipitous drop in success in coastal counties compared with previous years, portending negative impacts on population recruitment. As an apex predator and efficient scavenger, it is likely that bald eagles become infected through consumption of infected waterfowl. These results and similar reports of raptor mortality in Europe, Asia, and Africa, indicate a clear threat to raptor health. The possible long-term persistence of HP H5N1 IAV in North America poses an impending threat to bald eagle populations not only related to direct mortality but also decreased recruitment and warrants continued efforts to understand these potential impacts.

Mange is a contagious skin disease caused by different mite species affecting numerous domestic and wild animals, worldwide. This report details notoedric mange in an eastern cottontail (Sylvilagus floridanus) and in a marsh rabbit (Sylvilagus palustris) from Florida, USA. Clinical examination revealed similar gross lesions including poor nutritional condition, multifocal alopecia and hyperkeratosis. Skin scrapings from both rabbits revealed numerous subcutaneous mites identified as Notoedres centrifera, a species previously only associated with rodents, primarily squirrels. Mites from both rabbits were identified based on morphology and confirmed by sequencing the internal transcribed spacer-2 (ITS-2) region. These cases emphasize the need for continued surveillance and accurate diagnostic evaluation to determine the cause and characterization of the skin disease, while distinguishing it from other potential pathogens that may manifest similarly in rabbits, such as Notoedres cati, Sarcoptes scabiei or Psoroptes cuniculi.

In 1995, eight female pumas from Texas (Puma concolor stanleyana) were introduced into five areas in South Florida to remediate morphological and biomedical correlates of inbreeding depression that threatened the long-term survival of Florida panthers (P. c. coryi). Initial analysis of the results of this genetic introgression initiative has highlighted its success, including growth of the population, increased genetic heterozygosity, and reduced frequencies of physical traits associated with inbreeding, including cryptorchidism. However, panthers remain isolated from other puma populations and natural gene flow is not currently plausible. Using data collected from 65 panthers in the post-genetic introgression era (1998–2020), we assessed the benefits accrued to male panther reproductive parameters via this management initiative, the effect of genetic variables, and compared results to historic data. Microsatellite data were used to determine individual heterozygosity levels and qualify panther genetic ancestry as admixed (i.e., panthers descended from crosses with female Texas puma) or canonical (i.e., panthers that comprised most of the population prior to genetic introgression). A total of 47 admixed and 10 canonical panthers were identified; mean individual heterozygosity of canonical panthers (0.39) was significantly different from admixed panthers (0.63; t54 = 4.7, P-value < 0.001). Admixed panthers exhibited higher mean values for total testicular volume, structurally normal spermatozoa, and total sperm in comparison to canonical panthers. The progressive improvement of sperm metrics demonstrates that the benefits of genetic introgression continue to have a positive impact on the population >25 years since the release of the Texas pumas. However, the Florida panther population remains small, isolated, and vulnerable to deterministic and stochastic events that warrants continued monitoring to assess when additional genetic introgression initiatives will need to be implemented in the future.

Feline leukemia virus (FeLV) is a retrovirus that primarily affects domestic cats. Close interactions with domestic cats, including predation, can lead to the interspecific transmission of the virus to pumas, bobcats, or other feline species. Feline leukemia virus (FeLV) is a gammaretrovirus with horizontally transmitted and endogenous forms. Domestic cats are the primary reservoir species, but FeLV outbreaks in endangered Florida panthers and Iberian lynxes have resulted in mortalities. To assess prevalence and interspecific/intraspecific transmission, we conducted an extensive survey and phylogenetic analysis of FeLV infection in free-ranging pumas (n = 641) and bobcats (n = 212) and shelter domestic cats (n = 304). Samples were collected from coincident habitats across the United States between 1985 and 2018. FeLV infection was detected in 3.12% of the puma samples, 0.47% of the bobcat samples, and 6.25% of the domestic cat samples analyzed. Puma prevalence varied by location, with Florida having the highest rate of infection. FeLV env sequences revealed variation among isolates, and we identified two distinct clades. Both progressive and regressive infections were identified in cats and pumas. Based on the time and location of sampling and phylogenetic analysis, we inferred 3 spillover events between domestic cats and pumas; 3 puma-to-puma transmissions in Florida were inferred. An additional 14 infections in pumas likely represented spillover events following contact with reservoir host domestic cat populations. Our data provide evidence that FeLV transmission from domestic cats to pumas occurs widely across the United States, and puma-to-puma transmission may occur in genetically and geographically constrained populations.IMPORTANCE Feline leukemia virus (FeLV) is a retrovirus that primarily affects domestic cats. Close interactions with domestic cats, including predation, can lead to the interspecific transmission of the virus to pumas, bobcats, or other feline species. Some infected individuals develop progressive infections, which are associated with clinical signs of disease and can result in mortality. Therefore, outbreaks of FeLV in wildlife, including the North American puma and the endangered Florida panther, are of high conservation concern. This work provides a greater understanding of the dynamics of the transmission of FeLV between domestic cats and wild felids and presents evidence of multiple spillover events and infections in all sampled populations. These findings highlight the concern for pathogen spillover from domestic animals to wildlife but also identify an opportunity to understand viral evolution following cross-species transmissions more broadly.

Identifying drivers of transmission-especially of emerging pathogens-is a formidable challenge for proactive disease management efforts. While close social interactions can be associated with microbial sharing between individuals, and thereby imply dynamics important for transmission, such associations can be obscured by the influences of factors such as shared diets or environments. Directly-transmitted viral agents, specifically those that are rapidly evolving such as many RNA viruses, can allow for high-resolution inference of transmission, and therefore hold promise for elucidating not only which individuals transmit to each other, but also drivers of those transmission events. Here, we tested a novel approach in the Florida panther, which is affected by several directly-transmitted feline retroviruses. We first inferred the transmission network for an apathogenic, directly-transmitted retrovirus, feline immunodeficiency virus (FIV), and then used exponential random graph models to determine drivers structuring this network. We then evaluated the utility of these drivers in predicting transmission of the analogously transmitted, pathogenic agent, feline leukemia virus (FeLV), and compared FIV-based predictions of outbreak dynamics against empirical FeLV outbreak data. FIV transmission was primarily driven by panther age class and distances between panther home range centroids. FIV-based modeling predicted FeLV dynamics similarly to common modeling approaches, but with evidence that FIV-based predictions captured the spatial structuring of the observed FeLV outbreak. While FIV-based predictions of FeLV transmission performed only marginally better than standard approaches, our results highlight the value of proactively identifying drivers of transmission-even based on analogously-transmitted, apathogenic agents-in order to predict transmission of emerging infectious agents. The identification of underlying drivers of transmission, such as through our workflow here, therefore holds promise for improving predictions of pathogen transmission in novel host populations, and could provide new strategies for proactive pathogen management in human and animal systems.

Our work brings preliminary data on the diversity of microorganisms in Dobšinská and Demänovská ice caves (Slovakia). The caves are located at about 80 km from each other. Demänovská Ice Cave is part of the Demänovské caves system, located in the Demänovská Valley in the Low Tatras National Park. Dobšinská Ice Cave (969m asl), is part of the Stratená Cave System, located on the south-western edge of the Slovak Paradise National Park in the Spiš-Gemer karst. Both caves have been opened for the public during the last decades of the 19th century and are presently intensively visited by tourists [in 2014 alone, Demänovská ice cave was visited by 70,769 tourists (Nudziková 2018)]. There are few reports on the biocenosis of these two caves, the available data referring mainly to the invertebrate fauna (Papáč et al. 2019). In what concerns the microorganisms, up to our investigation, in focus have been only the fungi. Thus, in Dobšinská Ice Cave was reported the presence of species such as Botrytis cinerea and Aspergillus fumigatus isolated from bat guano (Nováková 2006) and the abundance of fungi outside and inside the cave was compared while from Demänovská Ice Cave, based on ITS sequences, Ogórek et al. (2018) reports on the phenotypic and genotypic diversity of airborne fungal spores. Our work complements these two studies adding data on the bacterial diversity from different parts of the two caves and depending on the substrate types (ice blocks and/or sediments). Using Sanger sequencing of the 16S amplicons from DNA isolated from pure bacterial cultures obtained from ice and/or sediments of Demänovská and Dobšinská Ice Caves, we identified 7 and respectively 12 species of bacteria. Surprisingly, only two species where common to both caves. In both Demänovská Ice Cave and Dobšinská Ice Cave the bacterial communities were dominated by Acinetobacter and Bacillus. Beside the identification of the main bacterial groups present in the underground habitats, Illumina sequencing enabled us to build genus heatmaps for the caves, allowing for comparision between various segments of each cave in relation with the specific climate features. When compared, the different types of samples (water, sediment, ice) showed different abundance of bacteria, ice diversity being greater than sediment/water diversity in both Dobšinska and Demänovská ice caves. The chemical composition of the samples (ice and water) is also discussed. This work is the first to bring insights on the bacterial diversity of Demänovská and Dobšinská Ice Caves and we appreciate what it opens new directions of research not only on the evolution of the microbial communities in relation to the climate changes occurred in past times, but also on how intense visiting affects the structures of the microorganisms communities.

Pathogen management strategies in wildlife are typically accompanied by an array of uncertainties such as the efficacy of vaccines or potential unintended consequences of interventions. In the context of such uncertainties, models of disease transmission can provide critical insight for optimizing pathogen management, especially for species of conservation concern. The endangered Florida panther experienced an outbreak of feline leukemia virus (FeLV) in 2002-04, and continues to be affected by this deadly virus. Ongoing management efforts aim to mitigate the effects of FeLV on panthers, but with limited information about which strategies may be most effective and efficient. We used a simulation-based approach to determine optimal FeLV management strategies in panthers. We simulated use of proactive FeLV management strategies (i.e., proactive vaccination) and several reactive strategies, including reactive vaccination and test-and-removal. Vaccination strategies accounted for imperfect vaccine-induced immunity, specifically partial immunity in which all vaccinates achieve partial pathogen protection. We compared the effectiveness of these different strategies in mitigating the number of FeLV mortalities and the duration of outbreaks. Results showed that inadequate proactive vaccination can paradoxically increase the number of disease-induced mortalities in FeLV outbreaks. These effects were most likely due to imperfect vaccine immunity causing vaccinates to serve as a semi-susceptible population, thereby allowing outbreaks to persist in circumstances otherwise conducive to fadeout. Combinations of proactive vaccination with reactive test-and-removal or vaccination, however, had a synergistic effect in reducing impacts of FeLV outbreaks, highlighting the importance of using mixed strategies in pathogen management. Synthesis and applications: Management-informed disease simulations are an important tool for identifying unexpected negative consequences and synergies among pathogen management strategies. In particular, we find that imperfect vaccine-induced immunity necessitates further consideration to avoid unintentionally worsening epidemics in some conditions. However, mixing proactive and reactive interventions can improve pathogen control while mitigating uncertainties associated with imperfect interventions.

Feral swine (Sus scrofa), an important prey species for the endangered Florida panther (Puma concolor coryi), is the natural host for pseudorabies virus (PRV). Prior to this study, PRV had been detected in just three panthers. To determine the effect of PRV on the panther population, we prospectively necropsied 199 panthers and retrospectively reviewed necropsy and laboratory findings, reexamined histology, and tested archived tissues using real-time PCR from 46 undiagnosed panther mortalities. Seven additional infections (two prospective, five retrospective) were detected for a total of 10 confirmed panther mortalities due to PRV. To further evaluate the effect of PRV, we categorized radio-collared (n=168) and uncollared panther mortalities (n=367) sampled from 1981 to 2018 based on the likelihood of PRV infection as confirmed, probable, suspected, possible, or unlikely/negative. Of 168 radio-collared panthers necropsied, PRV was the cause of death for between eight (confirmed; 4.8%) and 32 (combined confirmed, probable, suspected, and possible categories; 19.0%) panthers. The number of radio-collared panther mortalities due to PRV was estimated to be 15 (95% empirical limits: 12-19), representing 8.9% (confidence interval: 4.6-13.2%) of mortalities. Gross necropsy findings in 10 confirmed cases were nonspecific. Microscopic changes included slight to mild perivascular cuffing and gliosis (primarily in the brain stem), lymphoplasmacytic meningoencephalitis (cerebral cortex), and intranuclear inclusion bodies (adrenal medulla). The PRV glycoprotein C gene sequences from three positive panthers grouped with the sequence from a Florida feral swine. Our findings indicate that PRV may be an important and underdiagnosed cause of death in Florida panthers.

Feline foamy virus (FFV) is a retrovirus that has been detected in multiple feline species, including domestic cats (Felis catus) and pumas (Puma concolor). FFV results in persistent infection but is generally thought to be apathogenic. Sero-prevalence in domestic cat populations has been documented in several countries, but the extent of viral infections in nondomestic felids has not been reported. In this study, we screened sera from 348 individual pumas from Colorado, Southern California and Florida for FFV exposure by assessing sero-reactivity using an FFV anti-Gag ELISA. We documented a sero-prevalence of 78.6% across all sampled subpopulations, representing 69.1% in Southern California, 77.3% in Colorado, and 83.5% in Florida. Age was a significant risk factor for FFV infection when analyzing the combined populations. This high prevalence in geographically distinct populations reveals widespread exposure of puma to FFV and suggests efficient shedding and transmission in wild populations.

Determining parameters that govern pathogen transmission (such as the force of infection, FOI), and pathogen impacts on morbidity and mortality, is exceptionally challenging for wildlife. Vital parameters can vary, for example across host populations, between sexes and within an individual's lifetime. Feline immunodeficiency virus (FIV) is a lentivirus affecting domestic and wild cat species, forming species-specific viral-host associations. FIV infection is common in populations of puma (Puma concolor), yet uncertainty remains over transmission parameters and the significance of FIV infection for puma mortality. In this study, the age-specific FOI of FIV in pumas was estimated from prevalence data, and the evidence for disease-associated mortality was assessed. We fitted candidate models to FIV prevalence data and adopted a maximum likelihood method to estimate parameter values in each model. The models with the best fit were determined to infer the most likely FOI curves. We applied this strategy for female and male pumas from California, Colorado, and Florida. When splitting the data by sex and area, our FOI modeling revealed no evidence of disease-associated mortality in any population. Both sex and location were found to influence the FOI, which was generally higher for male pumas than for females. For female pumas at all sites, and male pumas from California and Colorado, the FOI did not vary with puma age, implying FIV transmission can happen throughout life; this result supports the idea that transmission can occur from mothers to cubs and also throughout adult life. For Florida males, the FOI was a decreasing function of puma age, indicating an increased risk of infection in the early years, and a decreased risk at older ages. This research provides critical insight into pathogen transmission and impact in a secretive and solitary carnivore. Our findings shed light on the debate on whether FIV causes mortality in wild felids like puma, and our approach may be adopted for other diseases and species. The methodology we present can be used for identifying likely transmission routes of a pathogen and also estimating any disease-associated mortality, both of which can be difficult to establish for wildlife diseases in particular.

Once on the brink of extinction, the Florida panther (Puma concolor coryi) has reoccupied parts of its extirpated range in southern Florida, USA over the past 20 years, which has largely been attributed to genetic restoration efforts initiated in 1995 to combat inbreeding depression and subsequent deleterious traits. Concurrent to the resurgence, an increase in documented livestock depredation events has heightened concern over human-panther conflicts. We examined 312 stomach content, scat, and feces (large intestine contents) samples collected 1989 to 2014 across the endemic range in southern Florida. We compared frequency of occurrence of prey items in samples by temporal (pre- and post-genetic restoration), geographic (north and south of 26 degrees 10.017' latitude), and demographic (age and sex) categories. We observed an apparent temporal shift in prey item occurrence in scats, where raccoon (Procyon lotor) occurrence increased while wild hog (Sus scrofa) occurrence decreased, whereas white-tailed deer (Odocoileus virginianus) occurrence appeared constant. Post-genetic restoration, we observed a geographic difference in panther prey, where white-tailed deer and raccoons were consumed more commonly in the southern part of the study area (characterized by lower soil quality and higher hydrological fluctuations), while wild hogs were consumed more frequently in the northern part of the study area. Neither sex nor age appeared to affect frequency of prey occurrence. Pets and livestock were not frequently found in the samples we examined. Overall, our results show shifts in panther diets both temporally and geographically; however, no notable changes in frequency of livestock found in panther diets were observed.