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Host population persistence in the face of introduced vector-borne diseases: Hawaii amakihi and avian malaria

The past quarter century has seen an unprecedented increase in the number of new and emerging infectious diseases throughout the world, with serious implications for human and wildlife populations. We examined host persistence in the face of introduced vector-borne diseases in Hawaii, where introduced avian malaria and introduced vectors have had a negative impact on most populations of Hawaiian forest birds for nearly a century. We studied birds, parasites, and vectors in nine study areas from 0 to 1,800 m on Mauna Loa Volcano, Hawaii from January to October, 2002. Contrary to predictions of prior work, we found that Hawaii amakihi (Hemignathus virens), a native species susceptible to malaria, comprised from 24.5% to 51.9% of the avian community at three low-elevation forests (55-270 m). Amakihi were more abundant at low elevations than at disease-free high elevations, and were resident and breeding there. Infection rates were 24-40% by microscopy and 55-83% by serology, with most infected individuals experiencing low-intensity, chronic infections. Mosquito trapping and diagnostics provided strong evidence for year-round local transmission. Moreover, we present evidence that Hawaii amakihi have increased in low elevation habitats on south-eastern Hawaii Island over the past decade. The recent emergent phenomenon of recovering amakihi populations at low elevations, despite extremely high prevalence of avian malaria, suggests that ecological or evolutionary processes acting on hosts or parasites have allowed this species to recolonize low-elevation habitats. A better understanding of the mechanisms allowing coexistence of hosts and parasites may ultimately lead to tools for mitigating disease impacts on wildlife and human populations.

Proceedings of the National Academy of Sciences of

Similar population dynamics before and after a chytridiomycosis outbreak in a tropical riparian amphibian species

Emerging infectious diseases can cause rapid, widespread host mortality, and the lack of demographic data before and after pathogen emergence complicates understanding mechanisms of host persistence. This challenge is further compounded by environmental conditions that influence host behavior, while driving pathogen growth and virulence. These interactions create complex disease outcomes that hinder predictions of when and how hosts endure pathogen outbreaks. Here, we analyzed 10 years of capture-mark-recapture data (2000–2014) spanning wet and dry seasons for male Espadarana prosoblepon in El Copé, Panama, encompassing a period before (2000–2004) and after (2010–2014) a Batrachochytrium dendrobatidis ( Bd ) outbreak using Jolly-Seber models. We found that post- Bd male E. prosoblepon population size (range in mean population size among primary periods = 136–225 individuals) was similar to pre- Bd population size (range in mean population size among primary periods = 201–242 individuals). Pre- Bd , average monthly survival probability in the wet season was 0.93 (95% credible interval [CI] = 0.90–0.96). Post- Bd , uninfected individuals had survival probability higher in the wet season (mean = 0.97; [95% CI = 0.95–0.98]) than the dry season (mean = 0.90 [95% CI = 0.84–0.94]), while survival probability for infected individuals decreased as a function of Bd infection intensity. Pre- Bd , mean monthly per-capita entry probability was 0.07 (95% CI = 0.05–0.10), and post- Bd , mean monthly per-capita entry probability was 0.06 (95% CI = 0.00–0.10). Lastly, infection probability during the wet season was lower (mean = 0.04 [95% CI = 0.03–0.05]) than the dry season (mean = 0.10 [95% CI = 0.05–0.15]), and recovery probability during the wet season was lower (mean = 0.19 [95% CI = 0.11–0.28]) than the dry season (mean = 0.54 [95% CI = 0.20–0.88]). Our findings suggest that survival probabilities of uninfected individuals, as well as per-capita entry probabilities, are similar pre- and post- Bd , leading to a stable and similar sized pre- Bd population. These results contribute to understanding disease dynamics and tropical amphibian ecology.

Ecosphere

Detection of tick-borne pathogen coinfections and coexposures to foot-and-mouth disease, brucellosis, and Q fever in selected wildlife from Kruger National Park, South Africa, and Etosha National Park, Namibia

Background: Although the rate of emerging infectious diseases that originate in wildlife has been increasing globally in recent decades, there is currently a lack of epidemiological data from wild animals. Methodology: We used serology to determine prior exposure to foot-and-mouth disease virus (FMDV), Brucella spp., and Coxiella burnetii and used genetic testing to detect blood-borne parasitic infections in the genera Ehrlichia , Anaplasma , Theileria , and Babesia from wildlife in two national parks, Kruger National Park (KNP), South Africa, and Etosha National Park (ENP), Namibia. Serum and whole blood samples were obtained from free-roaming plains zebra ( Equus quagga ), greater kudu ( Tragelaphus strepsiceros ), impala ( Aepyceros melampus ), and blue wildebeest ( Connochaetes taurinus ). Risk factors (host species, sex, and sampling park) for infection with each pathogen were assessed, as well as the prevalence and distribution of co-occurring infections. Results: In KNP 13/29 (45%; confidence interval [CI]: 26%–64%) kudus tested positive for FMD, but none of these reacted to SAT serotypes. For brucellosis, seropositive results were obtained for 3/29 (10%; CI: 2%–27%) kudu samples. Antibodies against C. burnetii were detected in 6/29 (21%; CI: 8%–40%) kudus, 14/21 (67%; CI: 43%–85%) impalas, and 18/39 (46%; CI: 30%–63%) zebras. A total of 28/28 kudus tested positive for Theileria spp. (100%; CI: 88%–100%) and 27/28 for Anaplasma/Ehrlichia spp. (96%; CI: 82%–100%), whereas 12/19 impalas (63%) and 2/39 zebra (5%) tested positive for Anaplasma centrale . In ENP, only 1/29 (3%; CI: 0%–18%) wildebeest samples tested positive for FMD. None of the samples tested positive for brucellosis, while C. burnetii antibodies were detected in 26/30 wildebeests (87%; CI: 69%–96%), 16/40 kudus (40%; CI: 25%–57%), and 26/26 plains zebras (100%; CI: 87%–100%). A total of 60% Anaplasma/Ehrlichia spp. and 35% Theileria/Babesia spp. in kudu and 37% wildebeest tested positive to Theileria sp. (sable), 30% to Babesia occultans , and 3%–7% to Anaplasma spp. The seroprevalence of Q fever was significantly higher in ENP, while Brucella spp., Anaplasma , Ehrlichia , Theileria , and Babesia species were significantly higher in KNP. Significant coinfections were also identified. Conclusion: This work provided baseline serological and molecular data on 40+ pathogens in four wildlife species from two national parks in southern Africa.

Etosha National Park, Kruger National Park

Snake fungal disease in North America: U.S. Geological Survey updates

Snake fungal disease (SFD) results from a skin infection that has been documented only in snakes. Historically, reports of snakes with skin infections of unknown origin have been sporadic. Recently, the number of reported cases of skin infections in snakes has increased substantially. This emerging infectious disease, confirmed in numerous species of snakes, is caused by the fungus Ophidiomyces ophiodiicola . As of August 2017, O. ophiodiicola has been detected in at least 23 States and one Canadian Province. However, researchers suspect that SFD may be more widely distributed than these documented cases suggest, because efforts to monitor the health of many snake populations are limited. Snake fungal disease may also be underreported in populations where it affects snakes infrequently or in species that develop less severe illness. Signs of SFD include crusted or ulcerated scales, nodules (that is, abnormal bumps) under the skin, and facial disfiguration that can be quite severe, leading to emaciation and death. Many snake populations are already in decline due to habitat loss and dwindling prey populations, and the recent emergence of SFD may accelerate this decline, causing certain species to disappear entirely from some locations.

Fact Sheet

Empirical evidence for effects of invasive American Bullfrogs on occurrence of native amphibians and emerging pathogens

Invasive species and emerging infectious diseases are two of the greatest threats to biodiversity. American Bullfrogs ( Rana [ Lithobates ] catesbeiana ), which have been introduced to many parts of the world, are often linked with declines of native amphibians via predation and spreading emerging pathogens such as amphibian chytrid fungus ( Batrachochytrium dendrobatidis [Bd]) and ranaviruses. Although many studies have investigated the potential role of bullfrogs in declines of native amphibians, analyses that account for shared habitat affinities and imperfect detection have found limited support for clear effects. Similarly, the role of bullfrogs in shaping the patch-level distribution of pathogens is unclear. We used eDNA methods to sample 233 sites in the southwestern USA and Sonora, Mexico (2016–2018) to estimate how presence of bullfrogs affects occurrence of 4 native amphibians, Bd, and ranaviruses. Based on 2-species, dominant-subordinate occupancy models fitted in a Bayesian context, federally threatened Chiricahua Leopard Frogs ( R. chiricahuensis ) and Western Tiger Salamanders ( Ambystoma mavortium ) were 8 times (32% vs. 4%) and 2 times (36% vs. 18%), respectively, less likely to occur at sites where bullfrogs occurred. Evidence for negative effects of bullfrogs on Lowland Leopard Frogs ( R. yavapaiensis ) and Northern Leopard Frogs ( R. pipiens ) was less clear, possibly because of smaller numbers of sites where these native species still occur and because bullfrogs often occur at lower densities in streams, the primary habitat for Lowland Leopard Frogs. At the community level, Bd was most likely to occur where bullfrogs co-occurred with native amphibians, which could increase risk to native species. Ranaviruses were estimated to occur at 33% of bullfrog-only sites, 10% of sites where bullfrogs and native amphibians co-occurred, and only 3% of sites where only native amphibians occurred. Of the 85 sites where we did not detect any of the 5 target amphibian species, we also did not detect Bd or ranaviruses; this suggests other hosts do not drive the distribution of these pathogens in our study area. Our results provide landscape-scale evidence that bullfrogs reduce occurrence of native amphibians and increase occurrence of pathogens, information that can clarify risks and aid the prioritization of conservation actions.

Ecological Applications

Optimal management decisions are robust to unknown dynamics in an amphibian metapopulation plagued by disease

Identifying conservation actions to recover threatened species can be challenging due to many ecological uncertainties. For example, major threats to a species' conservation are commonly known or suspected, but the specific impacts on population or metapopulation dynamics can be uncertain. This is frequently the case with emerging infectious diseases, including chytridiomycosis, a global driver of amphibian population declines caused by the fungal pathogens Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans . While these diseases are known to cause amphibian declines and extirpations, the mechanisms of their landscape-scale spread are still largely unknown. Such uncertainty can lead to inaction which may jeopardize timely recovery of a species. Decision analysis is a pragmatic approach to making transparent and defensible decisions while dealing with uncertainties. We investigated whether optimal actions aimed at recovering boreal toad ( Anaxyrus boreas boreas ) metapopulations in the southern Rocky Mountains are robust to the unknown dynamics of Bd spread using value of information and regret analyses. Value of information is a decision-analytic tool for calculating the value of new information in terms of performance on management objectives, while regret measures the cost of acting under incorrect information. We further conducted a stochastic sensitivity analysis to identify the relative effects of metapopulation parameters on system dynamics. We found optimal actions were robust to the unknown dynamics of Bd spread. While boreal toad breeding occurrence is highly sensitive to Bd distribution, the optimal decision is not. Resolving the unknown dynamics of Bd spread would lead to a minimal gain of less than one breeding toad subpopulation at the end of 50 years, given the currently available management actions. Applying a decision-analytic framework coupled with value of information and regret analyses can help frame how uncertainties affect decisions in a way that empowers decision makers.

Animal Conservation

Context-dependent representation of within- and between-model uncertainty: Aggregating probabilistic predictions in infectious disease epidemiology

Probabilistic predictions support public health planning and decision making, especially in infectious disease emergencies. Aggregating outputs from multiple models yields more robust predictions of outcomes and associated uncertainty. While the selection of an aggregation method can be guided by retrospective performance evaluations, this is not always possible. For example, if predictions are conditional on assumptions about how the future will unfold (e.g. possible interventions), these assumptions may never materialize, precluding any direct comparison between predictions and observations. Here, we summarize literature on aggregating probabilistic predictions, illustrate various methods for infectious disease predictions via simulation, and present a strategy for choosing an aggregation method when empirical validation cannot be used. We focus on the linear opinion pool (LOP) and Vincent average, common methods that make different assumptions about between-prediction uncertainty. We contend that assumptions of the aggregation method should align with a hypothesis about how uncertainty is expressed within and between predictions from different sources. The LOP assumes that between-prediction uncertainty is meaningful and should be retained, while the Vincent average assumes that between-prediction uncertainty is akin to sampling error and should not be preserved. We provide an R package for implementation. Given the rising importance of multi-model infectious disease hubs, our work provides useful guidance on aggregation and a deeper understanding of the benefits and risks of different approaches.

Journal of the Royal Society Interface

A call to action: Standardizing white-tailed deer harvest data in the Midwestern United States and implications for quantitative analysis and disease management

Recreational hunting has been the dominant game management and conservation mechanism in the United States for the past century. However, there are numerous modern-day issues that reduce the viability and efficacy of hunting-based management, such as fewer hunters, overabundant wildlife populations, limited access, and emerging infectious diseases in wildlife. Quantifying the drivers of recreational harvest by hunters could inform potential management actions to address these issues, but this is seldom comprehensively accomplished because data collection practices limit some analytical applications (e.g., differing spatial scales of harvest regulations and harvest data). Additionally, managing large-scale issues, such as infectious diseases, requires collaborations across management agencies, which is challenging or impossible if data are not standardized. Here we discuss modern issues with the prevailing wildlife management framework in the United States from an analytical point of view with a case study of white-tailed deer ( Odocoileus virginianus ) in the Midwest. We have four aims: (1) describe the interrelated processes that comprise hunting and suggest improvements to current data collections systems, (2) summarize data collection systems employed by state wildlife management agencies in the Midwestern United States and discuss potential for large-scale data standardization, (3) assess how aims 1 and 2 influence managing infectious diseases in hunted wildlife, and (4) suggest actionable steps to help guide data collection standards and management practices. To achieve these goals, Wisconsin Department of Natural Resources disseminated a questionnaire to state wildlife agencies (Illinois, Indiana, Iowa, Kentucky, Michigan, Minnesota, Missouri, Ohio, Wisconsin), and we report and compare their harvest management structures, data collection practices, and responses to chronic wasting disease. We hope our “call to action” encourages re-evaluation, coordination, and improvement of harvest and management data collection practices with the goal of improving the analytical potential of these data. A deeper understanding of the strengths and deficiencies of our current management systems in relation to harvest and management data collection methods could benefit the future development of comprehensive and collaborative management and research initiatives (e.g., adaptive management) for wildlife and their diseases.

Illinois, Indiana, Iowa, Kentucky, Michigan, Minne

Disease in a dynamic landscape: host behavior and wildfire reduce amphibian chytrid infection

Disturbances are often expected to magnify effects of disease, but these effects may depend on the ecology, behavior, and life history of both hosts and pathogens. In many ecosystems, wildfire is the dominant natural disturbance and thus could directly or indirectly affect dynamics of many diseases. To determine how probability of infection by the aquatic fungus Batrachochytrium dendrobatidis (Bd) varies relative to habitat use by individuals, wildfire, and host characteristics, we sampled 404 boreal toads ( Anaxyrus boreas boreas ) across Glacier National Park, Montana (USA). Bd causes chytridiomycosis, an emerging infectious disease linked with widespread amphibian declines, including the boreal toad. Probability of infection was similar for females and the combined group of males and juveniles. However, only 9% of terrestrial toads were infected compared to >30% of aquatic toads, and toads captured in recently burned areas were half as likely to be infected as toads in unburned areas. We suspect these large differences in infection reflect habitat choices by individuals that affect pathogen exposure and persistence, especially in burned forests where warm, arid conditions could limit Bd growth. Our results show that natural disturbances such as wildfire and the resulting diverse habitats can influence infection across large landscapes, potentially maintaining local refuges and host behaviors that facilitate evolution of disease resistance.

Montana

Disease‐structured N‐mixture models: A practical guide to model disease dynamics using count data

Obtaining inferences on disease dynamics (e.g., host population size, pathogen prevalence, transmission rate, host survival probability) typically requires marking and tracking individuals over time. While multistate mark–recapture models can produce high‐quality inference, these techniques are difficult to employ at large spatial and long temporal scales or in small remnant host populations decimated by virulent pathogens, where low recapture rates may preclude the use of mark–recapture techniques. Recently developed N ‐mixture models offer a statistical framework for estimating wildlife disease dynamics from count data. N ‐mixture models are a type of state‐space model in which observation error is attributed to failing to detect some individuals when they are present (i.e., false negatives). The analysis approach uses repeated surveys of sites over a period of population closure to estimate detection probability. We review the challenges of modeling disease dynamics and describe how N ‐mixture models can be used to estimate common metrics, including pathogen prevalence, transmission, and recovery rates while accounting for imperfect host and pathogen detection. We also offer a perspective on future research directions at the intersection of quantitative and disease ecology, including the estimation of false positives in pathogen presence, spatially explicit disease‐structured N ‐mixture models, and the integration of other data types with count data to inform disease dynamics. Managers rely on accurate and precise estimates of disease dynamics to develop strategies to mitigate pathogen impacts on host populations. At a time when pathogens pose one of the greatest threats to biodiversity, statistical methods that lead to robust inferences on host populations are critically needed for rapid, rather than incremental, assessments of the impacts of emerging infectious diseases.

Ecology and Evolution

Metagenomic sequencing sheds light on microbes putatively associated with pneumonia-related fatalities of white-tailed deer (Odocoileus virginianus)

With emerging infectious disease outbreaks in human, domestic and wild animal populations on the rise, improvements in pathogen characterization and surveillance are paramount for the protection of human and animal health, as well as the conservation of ecologically and economically important wildlife. Genomics offers a range of suitable tools to meet these goals, with metagenomic sequencing facilitating the characterization of whole microbial communities associated with emerging and endemic disease outbreaks. Here, we use metagenomic sequencing in a case-control study to identify microbes in lung tissue associated with newly observed pneumonia-related fatalities in 34 white-tailed deer ( Odocoileus virginianus ) in Wisconsin, USA. We identified 20 bacterial species that occurred in more than a single individual. Of these, only Clostridium novyi was found to substantially differ (in number of detections) between case and control sample groups; however, this difference was not statistically significant. We also detected several bacterial species associated with pneumonia and/or other diseases in ruminants ( Mycoplasma ovipneumoniae , Trueperella pyogenes , Pasteurella multocida , Anaplasma phagocytophilum , Fusobacterium necrophorum ); however, these species did not substantially differ between case and control sample groups. On average, we detected a larger number of bacterial species in case samples than controls, supporting the potential role of polymicrobial infections in this system. Importantly, we did not detect DNA of viruses or fungi, suggesting that they are not significantly associated with pneumonia in this system. Together, these results highlight the utility of metagenomic sequencing for identifying disease-associated microbes. This preliminary list of microbes will help inform future research on pneumonia-associated fatalities of white-tailed deer.

Microbial Genomics

The Amphibian Research and Monitoring Initiative (ARMI): 5-year report

The Amphibian Research and Monitoring Initiative (ARMI) is an innovative, multidisciplinary program that began in 2000 in response to a congressional directive for the Department of the Interior to address the issue of amphibian declines in the United States. ARMI’s formulation was cross-disciplinary, integrating U.S. Geological Survey scientists from Biology, Water, and Geography to develop a course of action (Corn and others, 2005a). The result has been an effective program with diverse, yet complementary, expertise. ARMI’s approach to research and monitoring is multiscale. Detailed investigations focus on a few species at selected local sites throughout the country; monitoring addresses a larger number of species over broader areas (typically, National Parks and National Wildlife Refuges); and inventories to document species occurrence are conducted more extensively across the landscape. Where monitoring is conducted, the emphasis is on an ability to draw statistically defensible conclusions about the status of amphibians. To achieve this objective, ARMI has instituted a monitoring response variable that has nationwide applicability. At research sites, ARMI focuses on studying species/environment interactions, determining causes of observed declines, and developing new techniques to sample populations and analyze data. Results from activities at all scales are provided to scientists, land managers, and policymakers, as appropriate. The ARMI program and the scientists involved contribute significantly to understanding amphibian declines at local, regional, national, and international levels. Within National Parks and National Wildlife Refuges, findings help land managers make decisions applicable to amphibian conservation. For example, the National Park Service (NPS) selected amphibians as a vital sign for several of their monitoring networks, and ARMI scientists provide information and assistance in developing monitoring methods for this NPS effort. At the national level, ARMI has had major exposure at a variety of meetings, including a dedicated symposium at the 2004 joint meetings of the Herpetologists’ League, the American Society of Ichthyologists and Herpetologists, and the Society for the Study of Amphibians and Reptiles. Several principal investigators have brought international exposure to ARMI through venues such as the World Congress of Herpetology in South Africa in 2005 (invited presentation by Dr. Gary Fellers), the Global Amphibian Summit, sponsored by the International Union for Conservation of Nature (IUCN) and Wildlife Conservation International, in Washington, D.C., 2005 (invited participation by Dr. P.S. Corn), and a special issue of the international herpetological journal Alytes focused on ARMI in 2004 (edited by Dr. C.K. Dodd, Jr.). ARMI research and monitoring efforts have addressed at least 7 of the 21 Threatened and Endangered Species listed by the U.S. Fish and Wildlife Service (California red-legged frog [Rana draytonii], Chiricahua leopard frog [R. chiricahuensis], arroyo toad [Bufo californicus], dusky gopher frog [Rana sevosa], mountain yellow-legged frog [R. muscosa], flatwoods salamander [Ambystoma cingulatum], and the golden coqui [Eleutherodactylus jasperi]), and 9 additional species of concern recognized by the IUCN. ARMI investigations have addressed time-sensitive research, such as emerging infectious diseases and effects on amphibians related to natural disasters like wildfire, hurricanes, and debris flows, and the effects of more constant, environmental change, like urban expansion, road development, and the use of pesticides. Over the last 5 years, ARMI has partnered with an extensive list of government, academic, and private entities. These partnerships have been fruitful and have assisted ARMI in developing new field protocols and analytic tools, in using and refining emerging technologies to improve accuracy and efficiency of data handling, in conducting amphibian disease, malformation, and environmental effects research, and in implementing a network of monitoring and research sites. Accomplishments from these endeavors include more than 40 publications on amphibian status and trends, nearly 100 publications on amphibian ecology and causes of declines, and over 30 methodological publications. Several databases have emerged as a result of ARMI and its partnerships; one, a digital atlas of ranges for all U.S. amphibian species, was used by the IUCN to display amphibian distribution maps in the Global Amphibian Assessment Project. Given the scope of ARMI and the panoply of projects, findings have had implications for policy. Investigations that demonstrate amphibian declines or illuminate causes of declines provide valuable information about habitat management, environmental effects, mechanisms for the spread of disease, and human/amphibian interfaces. This information has been made available to land managers, scientists, educators, Congress and other policymakers, and the public. The support afforded ARMI by Congress has been influential in the program’s development and success. The value of ARMI’s efforts will continue to increase as we are able to extend our studies spatially and temporally to answer critical questions with more confidence. We are using ARMI’s resources efficiently and continuing to develop innovative mechanisms for leveraging resources for maximum effectiveness during challenging financial times. This report is a 5-year retrospective of the structure, methodology, progress, and contributions to the broader scientific community that have resulted from this national USGS program. We evaluate ARMI’s success to date, with regard to the challenges faced by the program and the strengths that have emerged. We chart objectives for the next 5 years that build on current accomplishments, highlight areas meriting further research, and direct efforts to overcome existing weaknesses.

Scientific Investigations Report

Pathogenic lineage of Perkinsea associated with mass mortality of frogs across the United States

Emerging infectious diseases such as chytridiomycosis and ranavirus infections are important contributors to the worldwide decline of amphibian populations. We reviewed data on 247 anuran mortality events in 43 States of the United States from 1999–2015. Our findings suggest that a severe infectious disease of tadpoles caused by a protist belonging to the phylum Perkinsea might represent the third most common infectious disease of anurans after ranavirus infections and chytridiomycosis. Severe Perkinsea infections (SPI) were systemic and led to multiorganic failure and death. The SPI mortality events affected numerous anuran species and occurred over a broad geographic area, from boreal to subtropical habitats. Livers from all PCR-tested SPI-tadpoles (n = 19) were positive for the Novel Alveolate Group 01 (NAG01) of Perkinsea, while only 2.5% histologically normal tadpole livers tested positive (2/81), suggesting that subclinical infections are uncommon. Phylogenetic analysis demonstrated that SPI is associated with a phylogenetically distinct clade of NAG01 Perkinsea. These data suggest that this virulent Perkinsea clade is an important pathogen of frogs in the United States. Given its association with mortality events and tendency to be overlooked, the potential role of this emerging pathogen in amphibian declines on a broad geographic scale warrants further investigation.

Scientific Reports

Rapid risk assessment framework to estimate potential for spillback at human-wildlife interfaces

More than 60% of emerging infectious diseases of humans have a wildlife origin, and when these diseases spread through human populations to new geographical areas, there is a considerable risk of spillback from humans to wildlife species. Spillback events can have severe consequences for wildlife populations, where the disease may cause morbidity and mortality, and human populations, where the establishment in wildlife may lead to prolonged transmission or new exposures in humans. Mitigating these consequences requires identifying the key risk factors that lead to human–wildlife transmission events and implementing risk-reducing actions, a challenge given that cross-species transmission events are rare and often data deficient. To identify potential species and locations that are most likely to lead to these rare events, we developed a spatially explicit, rapid risk assessment framework that incorporates three components of the spillback process: wildlife susceptibility, wildlife exposure, and pathogen introduction pressure. To demonstrate the broad applicability of our framework, we conducted a rapid risk assessment on two recent emerging zoonotic pathogens in humans, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and mpox, to determine the relative spillback risk to wild mammalian species in the continental United States. The rapid risk assessment identified both species and locations with higher than expected spillback risk, providing managers and researchers with valuable information to prioritize surveillance and risk-mitigation actions. Our framework represents a rapid and flexible approach to assess the risks of spillback to wildlife populations during rapidly evolving zoonotic disease outbreaks.

Transboundary and Emerging Diseases

The scope and severity of white-nose syndrome on hibernating bats in North America

Assessing the scope and severity of threats is necessary for evaluating impacts on populations to inform conservation planning. Quantitative threat assessment often requires monitoring programs that provide reliable data over relevant spatial and temporal scales, yet such programs can be difficult to justify until there is an apparent stressor. Leveraging efforts of wildlife management agencies to record winter counts of hibernating bats, we collated data for 5 species from over 200 sites across 27 U.S. states and 2 Canadian provinces from 1995 to 2018 to determine the impact of white-nose syndrome (WNS), a deadly disease of hibernating bats. We estimated declines of winter counts of bat colonies at sites where the invasive fungus that causes WNS ( Pseudogymnoascus destructans ) had been detected to assess the threat impact of WNS. Three species undergoing species status assessment by the U.S. Fish and Wildlife Service ( Myotis septentrionalis , Myotis lucifugus , and Perimyotis subflavus ) declined by more than 90%, which warrants classifying the severity of the WNS threat as extreme based on criteria used by NatureServe. The scope of the WNS threat as defined by NatureServe criteria was large (36% of Myotis lucifugus range) to pervasive (79% of Myotis septentrionalis range) for these species. Declines for 2 other species ( Myotis sodalis and Eptesicus fuscus ) were less severe but still qualified as moderate to serious based on NatureServe criteria. Data-sharing across jurisdictions provided a comprehensive evaluation of scope and severity of the threat of WNS and indicated regional differences that can inform response efforts at international, national, and state or provincial jurisdictions. We assessed the threat impact of an emerging infectious disease by uniting monitoring efforts across jurisdictional boundaries and demonstrated the importance of coordinated monitoring programs, such as the North American Bat Monitoring Program (NABat), for data-driven conservation assessments and planning.

Conservation Biology

Design- and model-based recommendations for detecting and quantifying an amphibian pathogen in environmental samples

Accurate pathogen detection is essential for developing management strategies to address emerging infectious diseases, an increasingly prominent threat to wildlife. Sampling for free-living pathogens outside of their hosts has benefits for inference and study efficiency, but is still uncommon. We used a laboratory experiment to evaluate the influences of pathogen concentration, water type, and qPCR inhibitors on the detection and quantification of Batrachochytrium dendrobatidis ( Bd ) using water filtration. We compared results pre- and post-inhibitor removal, and assessed inferential differences when single versus multiple samples were collected across space or time. We found that qPCR inhibition influenced both Bd detection and quantification in natural water samples, resulting in biased inferences about Bd occurrence and abundance. Biases in occurrence could be mitigated by collecting multiple samples in space or time, but biases in Bd quantification were persistent. Differences in Bd concentration resulted in variation in detection probability, indicating that occupancy modeling could be used to explore factors influencing heterogeneity in Bd abundance among samples, sites, or over time. Our work will influence the design of studies involving amphibian disease dynamics and studies utilizing environmental DNA (eDNA) to understand species distributions.

Ecolology and Evolution

Selection, trans-species polymorphism, and locus identification of major histocompatibility complex class IIβ alleles of New World ranid frogs

Genes encoded by the major histocompatibility complex (MHC) play key roles in the vertebrate immune system. However, our understanding of the evolutionary processes and underlying genetic mechanisms shaping these genes is limited in many taxa, including amphibians, a group currently impacted by emerging infectious diseases. To further elucidate the evolution of the MHC in frogs (anurans) and develop tools for population genetics, we surveyed allelic diversity of the MHC class II ??1 domain in both genomic and complementary DNA of seven New World species in the genus Rana (Lithobates). To assign locus affiliation to our alleles, we used a "gene walking" technique to obtain intron 2 sequences that flanked MHC class II?? exon 2. Two distinct intron sequences were recovered, suggesting the presence of at least two class II?? loci in Rana. We designed a primer pair that successfully amplified an orthologous locus from all seven Rana species. In total, we recovered 13 alleles and documented trans-species polymorphism for four of the alleles. We also found quantitative evidence of selection acting on amino acid residues that are putatively involved in peptide binding and structural stability of the ??1 domain of anurans. Our results indicated that primer mismatch can result in polymerase chain reaction (PCR) bias, which influences the number of alleles that are recovered. Using a single locus may minimize PCR bias caused by primer mismatch, and the gene walking technique was an effective approach for generating single-copy orthologous markers necessary for future studies of MHC allelic variation in natural amphibian populations. ?? 2010 Springer-Verlag.

Immunogenetics

Effect of amphibian chytrid fungus (Batrachochytrium dendrobatidis) on apparent survival of frogs and toads in the western USA

Despite increasing interest in determining the population-level effects of emerging infectious diseases on wildlife, estimating effects of disease on survival rates remains difficult. Even for a well-studied disease such as amphibian chytridiomycosis (caused by the fungus Batrachochytrium dendrobatidis [Bd]), there are few estimates of how survival of wild hosts is affected. We applied hierarchical models to long-term capture-mark-recapture data (mean = 10.6 yrs, range = 6–15 yrs) from >5500 uniquely-marked individuals to estimate the effect of Bd on apparent survival of four threatened or endangered ranid frog species ( Rana draytonii , R. muscosa , R. pretiosa , R. sierrae ) at 14 study sites in California and Oregon (USA) and one bufonid toad ( Anaxyrus boreas ) at two study sites in Wyoming and Montana. Our models indicated that the presence of Bd on an individual reduced apparent survival of ranid frogs by ~6–15% depending on species and sex. The estimated difference between toads with and without Bd was 19% for the Montana population and 55% for the Wyoming population; however, the 95% Credible Interval of these estimates included zero. These results provide evidence for negative effects of Bd on survival in wild populations even in the absence of obvious die-offs. Determining what factors influence the magnitude of the effects of Bd on wildlife populations is an important next step toward identifying management actions. These estimates of Bd effects are important for understanding the extent and severity of disease, whether disease effects have changed over time, and for informing management actions.

California, Montana, Oregon, Wyoming