Abstract Background The continued spread of insecticide resistance in mosquito vectors of malaria and arboviral diseases may lead to operational failure of insecticide-based interventions if resistance is not monitored and managed efficiently. This study aimed to develop and validate a new WHO glass bottle bioassay method as an alternative to the WHO standard insecticide tube test to monitor mosquito susceptibility to new public health insecticides with particular modes of action, physical properties or both. Methods A multi-centre study involving 21 laboratories worldwide generated data on the susceptibility of seven mosquito species (Aedes aegypti, Aedes albopictus, Anopheles gambiae sensu stricto [An. gambiae s.s.], Anopheles funestus, Anopheles stephensi, Anopheles minimus and Anopheles albimanus) to seven public health insecticides in five classes, including pyrethroids (metofluthrin, prallethrin and transfluthrin), neonicotinoids (clothianidin), pyrroles (chlorfenapyr), juvenile hormone mimics (pyriproxyfen) and butenolides (flupyradifurone), in glass bottle assays. The data were analysed using a Bayesian binomial model to determine the concentration–response curves for each insecticide–species combination and to assess the within-bioassay variability in the susceptibility endpoints, namely the concentration that kills 50% and 99% of the test population (LC50 and LC99, respectively) and the concentration that inhibits oviposition of the test population by 50% and 99% (OI50 and OI99), to measure mortality and the sterilizing effect, respectively. Results Overall, about 200,000 mosquitoes were tested with the new bottle bioassay, and LC50/LC99 or OI50/OI99 values were determined for all insecticides. Variation was seen between laboratories in estimates for some mosquito species–insecticide combinations, while other test results were consistent. The variation was generally greater with transfluthrin and flupyradifurone than with the other compounds tested, especially against Anopheles species. Overall, the mean within-bioassay variability in mortality and oviposition inhibition were < 10% for most mosquito species-insecticide combinations. Conclusion Our findings, based on the largest susceptibility dataset ever produced on mosquitoes, showed that the new WHO bottle bioassay is adequate for evaluating mosquito susceptibility to new and promising public health insecticides currently deployed for vector control. The datasets presented in this study have been used recently by the WHO to establish 17 new insecticide discriminating concentrations (DCs) for either Aedes spp. or Anopheles spp. The bottle bioassay and DCs can now be widely used to monitor baseline insecticide susceptibility of wild populations of vectors of malaria and Aedes-borne diseases worldwide. Graphical abstract

Chronic wasting disease (CWD) is a fatal neurological disease of cervids caused by a misfolded protein with no vaccines or other cures to prevent infection and death. In the past decade, CWD has been recognized as one of the 10 most important challenges facing wildlife management. This paper evaluates the temporal and spatial patterns of CWD prevalence in southern Wisconsin white-tailed deer during the past 20 years. In most CWD areas, prevalence progresses from introduction to enzootic equilibrium in 15-20 years. In some monitoring areas, the disease grows at a much slower rate and remains below the equilibrium levels. Outbreaks are characterized by four distinct epizootic stages. Disease establishment (Stage 1) depends on the distance from a CWD focus. High rates of incidence and prevalence growth (Stage 2) are followed by slower growth as the relative number of susceptible deer declines (Stage 3). The rate of prevalence increase over time is higher in more forested ecoregions and in males (0.48 vs. 0.43) but similar in females (0.43 vs. 0.37) and yearlings (0.44 vs. 0.39). Habitat features, acting as surrogates for deer behavior and contact, may affect the rate of prevalence growth at a landscape (ecoregion) scale. Additionally, prevalence may be affected by deer management practices that favor higher deer abundance and more mature males. Finally, enzootic equilibrium (Stage 4) is higher in males (0.5), followed by females (0.36) and then yearlings (0.26). These equilibrium prevalence levels are high enough to have significant population impacts, reduce the abundance of mature males, and facilitate CWD spread by infected yearlings. Epizootic patterns suggest that CWD transmission has been frequency-dependent and primarily driven by direct contact with infected deer. Evidence for a meaningful change in the epizootic pattern from a 10-year management program to reduce deer abundance is lacking. The trajectory of CWD dynamics in Wisconsin suggests rapid growth in regional prevalence following introduction and increased spread across the landscape.

Pathogens can cause host extinction, affect population dynamics and influence natural selection. Host susceptibility to pathogens can vary by species, demographics and genetics which affect epizootic and population dynamics, ultimately determining population trends and evolution. Chronic wasting disease (CWD), a fatal neuro-degenerative prion disease of cervids, has varying host susceptibility conferred by polymorphisms of the prion protein gene (PRNP) at codon 96 for white-tailed deer Odocoileus virginianus. Deer with the homozygous Glycine allele (96GG) are most susceptible and a single Serine allele (96GS/96SS) reduces the risk of infection and mortality. We developed epizootiological models that demonstrate CWD infection and disease-associated mortality were higher for the more susceptible (96GG) genotype; and, infection was higher for males than females. We used population models to evaluate future shifts in genotype frequencies under alternative harvest and infection rate scenarios. Genetic shifts towards less susceptible genotypes were predicted as CWD prevalence increased during the course of an outbreak. This further increased CWD prevalence, and likely environmental contamination from prion shedding, due to longer incubation periods. Alternative harvest management strategies directly influenced CWD prevalence and spread, the rate of genetic selection and deer population growth. Synthesis and applications. We show that chronic wasting disease (CWD) transmission varied by sex, age class and PRNP genotype, and that CWD disease-associated mortality varied by genotype. Together, these forces lead to CWD-mediated genetic selection for a white-tailed deer population. We predict that genetic selection pressure increases when hunter harvest pressure is lowered, and conversely, increasing hunter harvest can reduce genetic selection rates of antlered deer. Our results support the control of CWD prevalence by aggressive harvest of adult males, because they have the highest infection rates. Our results have strong implications for evolution, disease ecology, geographical spread, disease mitigation and cervid population management.

Fleas are common ectoparasites of vertebrates worldwide and vectors of many pathogens causing disease, such as sylvatic plague in prairie dog colonies. Development of fleas is regulated by environmental conditions, especially temperature and relative humidity. Development rates are typically slower at low temperatures and faster at high temperatures, which are bounded by lower and upper thresholds where development is reduced. Prairie dogs and their associated fleas (mostly Oropsylla spp) live in burrows that moderate outside environmental conditions, remaining cooler in summer and warmer in winter. We found burrow microclimates were characterized by stable daily temperatures and high relative humidity, with temperatures increasing from spring through summer. We previously showed temperature increases corresponded with increasing off-host flea abundance. To evaluate how changes in temperature could affect future prairie dog flea development and abundance, we used development rates of O. montana (a species related to prairie dog fleas), determined how prairie dog burrow microclimates are affected by ambient weather, and combined these results to develop a predictive model. Our model predicts burrow temperatures and flea development rates will increase during the twenty-first century, potentially leading to higher flea abundance and an increased probability of plague epizootics if Y. pestis is present.

The development of models to elucidate the transmission pathways and dynamics of wildlife diseases remains challenging. Sylvatic plague, caused by the bacterium Yersinia pestis (Yp), is an infectious zoonotic disease that primarily affects wild rodents, including prairie dogs (Cynomys spp.) in North America. Proposed transmission pathways for Yp include flea bites, direct contacts between hosts, and environmental reservoirs (e.g. soil, carcasses). We developed a spatially explicit, agent-based model of Yp transmission to explore the effects of alternative transmission pathways, different disease initiation mechanisms (host or fleas), parameter uncertainty, and spatial structure of hosts. A particularly novel aspect of our model was the integration of ecological models with traditional disease models. Specifically, we used estimates from spatial capture-recapture models to generate data-driven spatial distributions, densities, and contact rates to capture the spatial structure of prairie dogs. We simulated similar to 9 million scenarios across a wide range of parameter values and conducted sensitivity analyses to determine the most influential parameters on the number of flea-days (sum of the mean number of fleas on hosts each day of the simulation), number of newly infected hosts per day, the time to depopulation (<20 prairie dogs remaining), and the proportion of the prairie dog population remaining at the end of the simulation (after 150 days). When including spatial structure, we found the probability of transmission via environmental sources of Yp (i.e. carcasses) had the greatest influence on model results when Yp infection was initiated in prairie dog hosts, rather than in fleas. Conversely, the mechanism of transmission by fleas to prairie dogs had the greatest influence on model results when Yp infection was initiated in fleas (i.e. via introduction by carnivores, a migrant prairie dog, or other mammalian host). Uncertainty in parameter estimates, particularly those related to the transmission pathways of Yp, continue to hamper efforts to realistically model plague dynamics in wild rodents. Our results elucidate the complexity of the flea-plague-prairie dog system and reiterate the importance of research on Yp transmission mechanisms to provide a full understanding of this disease. Our results also emphasize the importance of realistic estimates of spatial structure for exploring transmission dynamics of wildlife diseases and provide a framework for generating a data-driven description of spatial structure.

BACKGROUND Exposure to inorganic fertilizer is common for the major malaria vector Anopheles arabiensis, which is closely associated with agricultural activities. The aim of this study was to understand if insecticide susceptible and resistant individuals respond to fertilizer exposure in the same manner. Two laboratory strains, SENN, an insecticide susceptible strain, and SENN-DDT, an insecticide resistant strain selected strain selected from SENN, were used in this study. Both strains were exposed to one of three concentrations of a combination nitrogen-phosphorus-potassium (NPK) inorganic fertilizer, as well as nitrogenous (urea), phosphorus (superphosphate) and kaelic (potassium chloride, KCl) elemental fertilizer. The time to pupation was monitored, adult longevity was assessed and the insecticide tolerance of adults was determined. The effect of elemental fertilizers on ovipositioning site choice was also assessed. RESULTS For both strains, urea increased the number of eggs laid, while superphosphate resulted in a significant decrease in egg laying. Larval NPK exposure decreased the time to pupation in the SENN strain but not in SENN-DDT. Urea exposure increased the time to pupation in both strains, while KCl decreased the time to pupation in both strains. Larval NPK exposure only affected adult male longevity at high concentrations. Larval exposure to NPK and KCl resulted in increased insecticide tolerance in both strains, with variable efficacy from strain to strain. CONCLUSION Exposure to inorganic fertilizers has a greater effect on insecticide susceptible An. arabiensis as compared to resistant strains, where the primary advantage is increased insecticide tolerance. These data also demonstrate that larval fertilizer exposure can affect fecundity and fertility, and alter the life histories of adult An. arabiensis. (c) 2019 Society of Chemical Industry

Background Birds can maximize their reproductive success through careful selection of nest-sites. The ‘total-foliage’ hypothesis predicts that nests concealed in vegetation should have higher survival. We propose an additional hypothesis, the ‘predator proximity’ hypothesis, which states that nests placed farther from predators would have higher survival. We examined these hypotheses in the world’s southernmost forests of Navarino Island, in the Cape Horn Biosphere reserve, Chile (55°S). This island has been free of mammalian ground predators until recently, and forest passerines have been subject to depredation only by diurnal and nocturnal raptors. Methods During three breeding seasons (2014–2017), we monitored 104 nests for the five most abundant open-cup forest-dwelling passerines (Elaenia albiceps, Zonotrichia capensis, Phrygilus patagonicus, Turdus falcklandii, and Anairetes parulus). We identified nest predators using camera traps and assessed whether habitat characteristics affected nest-site selection and survival. Results Nest predation was the main cause of nest failure (71% of failed nests). Milvago chimango was the most common predator, depredating 13 (87%) of the 15 nests where we could identify a predator. By contrast, the recently introduced mammal Neovison vison, the only ground predator, depredated one nest (7%). Species selected nest-sites with more understory cover and taller understory, which according to the total-foliage hypothesis would provide more concealment against both avian and mammal predators. However, these variables negatively influenced nest survival. The apparent disconnect between selecting nest-sites to avoid predation and the actual risk of predation could be due to recent changes in the predator assemblage driven by an increased abundance of native M. chimango associated with urban development, and/or the introduction of exotic mammalian ground predators to this island. These predator assemblage changes could have resulted in an ecological trap. Further research will be needed to assess hypotheses that could explain this mismatch between nest-site selection and nest survival.

Sylvatic plague, caused by the bacterium Yersinia pestis and transmitted by fleas, occurs in prairie dogs of the western United States. Outbreaks can devastate prairie dog communities, often causing nearly 100% mortality. Three competent flea vectors, prairie dog specialists Oropsylla hirsuta and O. tuberculata, and generalist Pulex simulans, are found on prairie dogs and in their burrows. Fleas are affected by climate, which varies across the range of black-tailed prairie dogs (Cynomys ludovicianus), but these effects may be ameliorated somewhat due to the burrowing habits of prairie dogs. Our goal was to assess how temperature and precipitation affect off-host flea abundance and whether relative flea abundance varied across the range of black-tailed prairie dogs. Flea abundance was measured by swabbing 300 prairie dog burrows at six widely distributed sites in early and late summer of 2016 and 2017. Relative abundance of flea species varied among sites and sampling sessions. Flea abundance and prevalence increased with monthly mean high temperature and declined with higher winter precipitation. Predicted climate change in North America will likely influence flea abundance and distribution, thereby impacting plague dynamics in prairie dog colonies.

First posted October 23, 2019 Revised November 20, 2019 For additional information, contact: Associate Director, EcosystemsU.S. Geological Survey954 National Center12201 Sunrise Valley DriveReston, VA 20192 Chronic wasting disease (CWD) is the only transmissible spongiform encephalopathy, a class of invariably fatal neurodegenerative mammalian diseases associated with a misfolded cellular prion protein found in wild free-ranging animals. Because it has a long incubation period, affected animals in Cervidae (the deer family; referred to as “cervids”) may not show signs of disease for several years. While signs are not specific to CWD, affected cervids (deer, elk, moose, and reindeer) show changes in appearance (such as progressive weight loss) and changes in behavior such as stumbling, tremors, and teeth grinding. CWD can be transmitted by direct contact or through a contaminated environment. The causative prion agent is highly resistant to degradation.In recent decades, CWD has transitioned from a novel, obscure prion disease of cervids with limited geographical distribution, to a disease that poses substantial ecological, agricultural, and economic risks across large regions of North America. Since its discovery in free-ranging elk and deer populations in the western United States in the 1980s, CWD has been reported in captive or free-ranging cervid populations in 26 States, 3 Canadian Provinces, the Republic of South Korea, Finland, Sweden, and Norway. In addition, the proportion of CWD-infected animals is increasing in many areas where the disease is already established. In some heavily affected areas, total cervid numbers have decreased over time due to CWD, which suggests that these cervid populations may not be sustainable in the long-term.The U.S. Geological Survey (USGS) conducts wildlife disease surveillance and research to support management of CWD-affected species and their habitats. The scientific information is relevant to governmental agencies that manage wildlife and their habitats including the U.S. Fish and Wildlife Service, the National Park Service, the U.S. Department of Agriculture, and other Federal, State, and Tribal agencies as well as conservation partners (non-governmental organizations, businesses, and private landowners). Each project description in this report (1–30) includes the non-USGS collaborators (Federal, State, Tribal agencies, universities) and a USGS point of contact (principal investigator). If there are USGS publications associated with the project, a publication list is provided at the end of each project description.

Avian malaria has played a significant role in causing extinctions, population declines, and limiting the elevational distribution of Hawaiian honeycreepers. Most threatened and endangered honeycreepers only exist in high-elevation forests where the risk of malaria infection is limited. Because Culex mosquito vectors and avian malaria dynamics are strongly influenced by temperature and rainfall, future climate change is predicted to expand malaria infection to high-elevation forests and intensify malaria infection at lower elevations, likely resulting in future extinctions and loss of avian biodiversity in Hawaii. Novel, landscape-level mosquito control strategies are promising, but are logistically challenging and require costly long-term efforts. As an alternative or supplemental strategy, we evaluated the potential of releasing a gene-edited malaria-resistant honeycreeper (Iiwi, Drepanis coccinea) in Hawaiian rainforests; a strategy known as facilitated adaptation. While this approach also has significant technical challenges and costs, it may offer a more permanent solution to increasing malaria threats. If malaria-resistant honeycreepers can be developed, facilitated adaptation may provide a practical strategy for the reestablishment of abundant avian populations in Hawaiian forests. A successful strategy could be the release of malaria-resistant Iiwi in mid-elevation forests where development of a resistant population has the best chance of success. Establishment of a resistant Iiwi population could provide a source for dispersal and development of resistant populations in high-elevation forests and a permanent source of resistant individuals for translocation to other vulnerable areas.

Oral vaccination is an emerging management strategy to reduce the prevalence of high impact infectious diseases within wild animal populations. Plague is a flea-borne zoonosis of rodents that often decimates prairie dog (Cynomys spp.) colonies in the western USA. Recently, an oral sylvatic plague vaccine (SPV) was developed to protect prairie dogs from plague and aid recovery of the endangered black-footed ferret (Mustela nigripes). Although oral vaccination programs are targeted toward specific species, field distribution of vaccine-laden baits can result in vaccine uptake by non-target animals and unintended indirect effects. We assessed the impact of SPV on non-target rodents at paired vaccine and placebo-treated prairie dog colonies in four US states from 2013 to 2015. Bait consumption by non-target rodents was high (70.8%, n = 3113), but anti-plague antibody development on vaccine plots was low (23.7%, n = 266). In addition, no significant differences were noted in combined deer mice (Peromyscus maniculatus) and western harvest mouse (Reithrodontomys megalotis) abundance or community evenness and richness of non-target rodents between vaccine-treated and placebo plots. In our 3-year field study, we could not detect a significant positive or negative effect of SPV application on non-target rodents.

Chronic wasting disease (CWD) is a fatal neurodegenerative disease of deer, elk, moose, and reindeer (cervids) caused by misfolded prion proteins. The disease has been reported across North America and recently discovered in northern Europe. Transmission of CWD in wild cervid populations can occur through environmental routes, but limited ability to detect prions in environmental samples has prevented the identification of potential transmission “hot spots”. We establish widespread CWD prion contamination of mineral licks used by free-ranging cervids in an enzootic area in Wisconsin, USA. We show mineral licks can serve as reservoirs of CWD prions and thus facilitate disease transmission. Furthermore, mineral licks attract livestock and other wildlife that also obtain mineral nutrients via soil and water consumption. Exposure to CWD prions at mineral licks provides potential for cross-species transmission to wildlife, domestic animals, and humans. Managing deer use of mineral licks warrants further consideration to help control outbreaks of CWD.

1. Surveillance is critical for early detection of emerging and re-emerging infectious diseases. Weighted surveillance leverages heterogeneity in infection risk to increase sampling efficiency. 2. Here, we apply a Bayesian approach to estimate weights for 16 surveillance classes of white-tailed deer in Wisconsin, USA, relative to hunter-harvested yearling males. We used these weights to conduct a surveillance programme for detecting chronic wasting disease (CWD) in white-tailed deer at Shenandoah National Park (SHEN) in Virginia, USA. 3. Generally, for surveillance, risk of infection increased with age and was greater in males. Clinical suspect deer had the highest risk, with weight estimates of 33.33 and 9.09 for community-reported and hunter-reported suspect deer, respectively. Fawns had the lowest risk with an estimated weight of 0.001. 4. We used surveillance weights for Wisconsin deer to determine sampling effort required to detect a CWD-positive case in SHEN if prevalence in yearling males >= 0.025. The sampling required to detect CWD was 37-91 adult deer, depending on the adult male: female ratio in the surveillance stream. We collected rectal biopsies from 49 female and 21 male adult deer, and 10 additional samples from vehicle-killed deer. CWD was not detected and we concluded with 95% probability that prevalence in the reference population (yearling males) was between 0.0% and 3.6%. 5. Synthesis and applications. Our approach allows managers to estimate relative surveillance weights for different host classes and quantify limits of disease detection in real time when only a sample of animals from a population can be tested, resulting in considerable cost savings for agencies performing wildlife disease detection surveillance. Additionally, it provides a rigorous means of estimating prevalence limits when a disease/pathogen is not detected in a sample set. It is therefore applicable to other wildlife, domestic animal and human disease systems, which can be characterized by surveillance classes with heterogeneous probability of infection. This methodology is also extendable to other disciplines such as invasive species, environmental toxicology, and generally, any ecological question seeking to efficiently use scarce financial and human resources to maximize the detection probability of a rare event.

Despite the purported role of avian pox (Avipoxvirus spp.) in the decline of endemic Hawaiian birds, few studies have been conducted on the dynamics of this disease, its impact on free-living avian populations, or its interactions with avian malaria (Plasmodium relictum). We conducted four longitudinal studies of 3-7 yr in length and used generalized linear models to evaluate cross-sectional prevalence of active pox infection and individuals with healed deformities that had recovered from pox. Our goal was to understand how species, season, elevation, malaria infection, and other biological characteristics influenced pox infection in Modified Letter Turned CommaApapane, HawaiModified Letter Turned Commai Modified Letter Turned CommaAmakihi, Modified Letter Turned CommaIModified Letter Turned Commaiwi, and Japanese White-eye across low-, mid-, and high-elevation forests on the island of HawaiModified Letter Turned Commai. We also used multi-state capture-recapture (longitudinal) models to estimate pox infection rates, recovery rates, and potential pox-related mortality. Pox infection rates were typically highest in low-elevation forests, followed by mid-elevation forests, and lowest in high-elevation forests. We also found seasonal changes in pox prevalence throughout the annual cycle; typically increasing from spring through summer, peaking in fall, and declining in winter. These seasonal changes occurred in low- and mid-elevation forests, but not in high elevations where pox infection was low. Seasonal and elevation patterns of pox infection are like those for avian malaria, strongly implicating mosquito vectors, rather than other biting arthropods or contact transmission, as the primary source of transmitting both diseases. Most native Hawaiian birds recovered from pox infection within 6 months; frequently without permanent lesions. Contrary to our expectations, we found no direct evidence that pox is a substantial mortality factor in any of the three native bird species we studied. Birds with chronic malaria infection were more likely to have both active pox infection and healed pox lesions suggesting a synergistic interaction that may influence the evolution of pox virulence. Because pox infection can be assessed visually, and birds have a high recovery rate, this disease may be a sensitive indicator of the seasonal and annual risk of transmission of malaria in HawaiModified Letter Turned Commai.

Avian malaria, transmitted by Culex quinquefasciatus mosquitoes in the Hawaiian Islands, has been a primary contributor to population range limitations, declines, and extinctions for many endemic Hawaiian honeycreepers. Avian malaria is strongly influenced by climate; therefore, predicted future changes are expected to expand transmission into higher elevations and intensify and lengthen existing transmission periods at lower elevations, leading to further population declines and potential extinction of highly susceptible honeycreepers in mid- and high-elevation forests. Based on future climate changes and resulting malaria risk, we evaluated the viability of alternative conservation strategies to preserve endemic Hawaiian birds at mid and high elevations through the 21st century. We linked an epidemiological model with three alternative climatic projections from the Coupled Model Intercomparison Project to predict future malaria risk and bird population dynamics for the coming century. Based on climate change predictions, proposed strategies included mosquito population suppression using modified males, release of genetically modified refractory mosquitoes, competition from other introduced mosquitoes that are not competent vectors, evolved malaria-tolerance in native honeycreepers, feral pig control to reduce mosquito larval habitats, and predator control to improve bird demographics. Transmission rates of malaria are predicted to be higher than currently observed and are likely to have larger impacts in high-elevation forests where current low rates of transmission create a refuge for highly-susceptible birds. As a result, several current and proposed conservation strategies will be insufficient to maintain existing forest bird populations. We concluded that mitigating malaria transmission at high elevations should be a primary conservation goal. Conservation strategies that maintain highly susceptible species like Iiwi (Drepanis coccinea) will likely benefit other threatened and endangered Hawai’i species, especially in high-elevation forests. Our results showed that mosquito control strategies offer potential long-term benefits to high elevation Hawaiian honeycreepers. However, combined strategies will likely be needed to preserve endemic birds at mid elevations. Given the delay required to research, develop, evaluate, and improve several of these currently untested conservation strategies we suggest that planning should begin expeditiously.

Our ability to infer unobservable disease-dynamic processes such as force of infection (infection hazard for susceptible hosts) has transformed our understanding of disease transmission mechanisms and capacity to predict disease dynamics. Conventional methods for inferring FOI estimate a time-averaged value and are based on population-level processes. Because many pathogens exhibit epidemic cycling and FOI is the result of processes acting across the scales of individuals and populations, a flexible framework that extends to epidemic dynamics and links within-host processes to FOI is needed. Specifically, within-host antibody kinetics in wildlife hosts can be short-lived and produce patterns that are repeatable across individuals, suggesting individual-level antibody concentrations could be used to infer time since infection and hence FOI. Using simulations and case studies (influenza A in lesser snow geese and Yersinia pestis in coyotes), we argue that with careful experimental and surveillance design, the population-level FOI signal can be recovered from individual-level antibody kinetics, despite substantial individual-level variation. In addition to improving inference, the cross-scale quantitative antibody approach we describe can reveal insights into drivers of individual-based variation in disease response, and the role of poorly understood processes such as secondary infections, in population-level dynamics of disease.

Altitudinal movement by tropical birds to track seasonally variable resources can move them from protected areas to areas of increased vulnerability. In Hawai?i, historical reports suggest that many Hawaiian honeycreepers such as the Iiwi (Drepanis coccinea) once undertook seasonal migrations, but the existence of such movements today is unclear. Because Hawaiian honeycreepers are highly susceptible to avian malaria, currently minimal in high-elevation forests, understanding the degree to which honeycreepers visit lower elevation forests may be critical to predict the current impact of malaria on population dynamics and how susceptible bird populations may respond to climate change and mitigation scenarios. Using radio telemetry data, we demonstrate for the first time that a large fraction of breeding adult and juvenile Iiwi originating from an upper-elevation (1,920m) population at Hakalau Forest National Wildlife Refuge exhibit post-breeding movements well below the upper elevational limit for mosquitoes. Bloom data suggest seasonal variation in floral resources is the primary driver of seasonal movement for Iiwi. To understand the demographic implications of such movement, we developed a spatial individual-based model calibrated using previously published and original data. ?Iiwi dynamics were simulated backward in time, to estimate population levels in the absence of avian malaria, and forward in time, to assess the impact of climate warming as well as two potential mitigation actions. Even in disease-free refuge' populations, we found that breeding densities failed to reach the estimated carrying capacity, suggesting the existence of a seasonal migration load as a result of travel to disease-prevalent areas. We predict that Iiwi may be on the verge of extinction in 2100, with the total number of pairs reaching only +/- 0.2-12.3% of the estimated pre-malaria density, based on an optimistic climate change scenario. The probability of extinction of Iiwi populations, as measured by population estimates for 2100, is strongly related to their estimated migration propensity. Long-term conservation strategies likely will require a multi-pronged response including a reduction of malaria threats, habitat restoration and continued landscape-level access to seasonally variable nectar resources.

Chronic wasting disease (CWD) is the only naturally occurring transmissible spongiform encephalopathy affecting free-ranging wildlife populations. Transmission of CWD occurs by direct contact or through contaminated environments; however, little is known about the temporal patterns of CWD prion excretion and shedding in wild cervids. We tested the urine and faeces of three species of captive cervids (elk, mule and white-tailed deer) at 6, 12, 18 and 24 months after oral inoculation to evaluate the temporal, species- and genotype-specific factors affecting the excretion of CWD prions. Although none of the animals exhibited clinical signs of CWD during the study, we determined that all three cervid species were excreting CWD prions by 6 months post-inoculation. Faecal samples were consistently positive for CWD prions for all three cervid species (88 %), and were more likely to be positive than urine samples (28 %). Cervids with genotypes encoding for the prion protein (PRNP) that were considered to be more susceptible to CWD were more likely to excrete CWD prions (94 %) than cervids with genotypes considered to be less susceptible (64 %). All cervids with CWD prions in their urine also had positive faeces (n=5), but the converse was not true. Our study is the first to demonstrate CWD prion excretion in urine by asymptomatic elk and mule deer. Our results indicate that the excretion of CWD prions in faeces and, to a lesser extent, urine may provide an important avenue for depositing prions in the environment.