DISTRIBUTION, DIET, AND PREVALENCE OF AMPHIBIAN CHYTRID FUNGUS IN NON-NATIVE AMERICAN BULLFROGS (LITHOBATES CATESBEIANUS) AT THE VALENTINE NATIONAL WILDLIFE REFUGE, NEBRASKA, USA

American Bullfrogs (Lithobates catesbeianus) have been widely introduced beyond their native range in North America and can negatively affect organisms in wetland environments via a suite of mechanisms including interspecific interactions and disease transmission. Bullfrogs were introduced to the Valentine National Wildlife Refuge in the Sandhill Region of Nebraska, USA, yet little is known regarding their distribution, abundance, and potential effects on other vertebrates in the Refuge. Surveys in 1991-1992 documented bullfrogs in only one lake by the Refuge headquarters, and anecdotal historical observations indicated that bullfrogs were present primarily in lakes open to public fishing. In 2012, we determined the distribution of bullfrogs across the Refuge, examined their diets, and sampled them for the occurrence of the pathogenic fungus, Batrachochytrium dendrobatidis. We documented that bullfrogs were almost ubiquitous across the Refuge lakes and wetlands. From diet analyses, we observed that adult bullfrogs consumed several vertebrate species on the Refuge including: a Blanding’s Turtle (Emydoidea blandingii), small bullfrogs, an unidentifiable frog species, and numerous invertebrates, including crayfish. The amphibian chytrid fungus was prevalent on the refuge with 73.7% of bullfrogs testing positive for the fungus in early June and 6.3% in late June-July. Preliminary data indicate that bullfrogs likely have already affected interspecific interactions with native amphibians via predation, competition, and/or disease transmission. Bullfrogs likely cannot be eradicated from the Refuge, but expanding the season of harvest of bullfrogs might reduce their abundance, which may benefit native amphibians and reptiles

Spatial Variation of False Map Turtle (Graptemys pseudogeographica) Bacterial Microbiota in the Lower Missouri River, United States

Turtle populations around the world are continually confronted with changing environments that affect their ecology and conservation status. Among freshwater turtles, population dynamics are thought to be mediated by complex yet often cryptic causes. One recent direction of focus in addressing these causes is the turtle-associated microbiota. In turtles, the gut- associated microbiota is of exceptional interest due to its continual association with host species under changing conditions. Diet-based fluctuations and changes in microbial diversity may correspond to varying external environments at both the individual and population level. Environmental responses are of particular interest due to the anthropogenic changes that may underlie them. Pollutants, disruption of climatic patterns, and habitat fragmentation all have the potential to affect turtle-associated microbiota and subsequent population and species conservation. To better understand potential human-induced changes, the diversity of turtle-associated microbiota over local spatial gradients must be better understood. We examined microbial community alpha- and beta-diversity among 30 adult False Map Turtles (Graptemys pseudogeographica) at three sites within the lower Missouri River, United States. Our results indicate significant microbial community centroid differences among sites (beta-diversity), which are likely mediated by various local environmental factors. Such factors will have to be carefully considered in any future attribution of anthropogenic determinants on turtle-associated microbiota as it relates to turtle population dynamics

Host species composition influences infection severity among amphibians in the absence of spillover transmission

Wildlife epidemiological outcomes can depend strongly on the composition of an ecological community, particularly when multiple host species are affected by the same pathogen. However, the relationship between host species richness and disease risk can vary with community context and with the degree of spillover transmission that occurs among co‐occurring host species. We examined the degree to which host species composition influences infection by Batrachochytrium dendrobatidis (Bd), a widespread fungal pathogen associated with amphibian population declines around the world, and whether transmission occurs from one highly susceptible host species to other co‐occurring host species. By manipulating larval assemblages of three sympatric amphibian species in the laboratory, we characterized the relationship between host species richness and infection severity, whether infection mediates growth and survivorship differently across various combinations of host species, and whether Bd is transmitted from experimentally inoculated tadpoles to uninfected tadpoles. We found evidence of a dilution effect where Bd infection severity was dramatically reduced in the most susceptible of the three host species (Anaxyrus boreas). Infection also mediated survival and growth of all three host species such that the presence of multiple host species had both positive (e.g., infection reduction) and negative (e.g., mortality) effects on focal species. However, we found no evidence that Bd infection is transmitted by this species. While these results demonstrate that host species richness as well as species identity underpin infection dynamics in this system, dilution is not the product of reduced transmission via fewer infectious individuals of a susceptible host species. We discuss various mechanisms, including encounter reduction and antagonistic interactions such as competition and opportunistic cannibalism that may act in concert to mediate patterns of infection severity, growth, and mortality observed in multihost communities.There are many ways in which infection can be influenced by species diversity. Here we show experimentally that the interactions between species in a multi‐host amphibian community drive the severity of infection by the amphibian chytrid fungus. We find no evidence that infection is transmitted between two host species in our study, suggesting that spillover infection is not a cause of dilution effects in this system.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/111214/1/ece31385.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/111214/2/ece31385-sup-0001-FigureS1.pd

Integrative behavioral ecotoxicology: bringing together fields to establish new insight to behavioral ecology, toxicology, and conservation

The fields of behavioral ecology, conservation science, and environmental toxicology individually aim to protect and manage the conservation of wildlife in response to anthropogenic stressors, including widespread anthropogenic pollution. Although great emphasis in the field of toxicology has been placed on understanding how single pollutants affect survival, a comprehensive, interdisciplinary approach that includes behavioral ecology is essential to address how anthropogenic compounds are a risk for the survival of species and populations in an increasingly polluted world. We provide an integrative framework for behavioral ecotoxicology using Tinbergen\u27s four postulates (causation and mechanism, development and ontogeny, function and fitness, and evolutionary history and phylogenetic patterns). The aims of this review are: 1) to promote an integrative view and re-define the field of integrative behavioral ecotoxicology; 2) to demonstrate how studying ecotoxicology can promote behavior research; and 3) to identify areas of behavioral ecotoxicology that require further attention to promote the integration and growth of the field

Prevalence and Distribution of Ranavirus in Amphibians From

Several infectious diseases are threatening amphibian species worldwide and have resulted in massmortality events across the globe. An emerging group of viral pathogens (ranaviruses) are documented to cause die-offs in amphibian populations worldwide, including in several regions of the U.S. Unfortunately, large gaps remain in our understanding of the distribution of this systemic pathogen in the U.S., including within the state of Oklahoma. To address this gap in our understanding, we carried out surveys of this infectious pathogen across 14 sites in seven southeastern Oklahoma counties in spring 2015, screening 17 amphibian species from this region. Using liver and tail tissue samples collected from individual amphibians, we screened for the presence and infection load of ranavirus. Of the 390 samples, 84 (21.5%) tested positive for ranavirus, with infection prevalence varying among species surveyed. Notably, the family Bufonidae had no samples that tested positive for ranavirus, whereas the remaining families had an infection prevalence ranging from 14–50%. Despite an overall infection prevalence of 21.5%, we detected no clinical signs of ranavirosis and all sampled individuals appeared outwardly healthy. These results provide data on the geographic and host distribution of ranavirus in southeastern Oklahoma, as well as the first documented cases of the pathogen in three species of anurans: Gastrophryne carolinensis (Eastern Narrow-mouthed Toad), G. olivacea (Western Narrow-mouthed Toad), and Pseudacris fouquettei (Cajun Chorus Frog). With widespread ranavirus infection, there is potential for transmission from abundant, widespread species to more vulnerable, state-threatened amphibians

Genotyping and differential bacterial inhibition of Batrachochytrium dendrobatidis in threatened amphibians in Costa Rica

Amphibians have declined around the world in recent years, in parallel with the emergence of an epidermal disease called chytridiomycosis, caused by the chytrid fungus Batrachochytrium dendrobatidis (Bd). This disease has been associated with mass mortality in amphibians worldwide, including in Costa Rica, and Bd is considered an important contributor to the disappearance of this group of vertebrates. While many species are susceptible to the disease, others show tolerance and manage to survive infection with the pathogen. We evaluated the pathogen Bd circulating in Costa Rica and the capacity of amphibian skin bacteria to inhibit the growth of the pathogen in vitro. We isolated and characterized – genetically and morphologically – several Bd isolates from areas with declining populations of amphibians. We determined that the circulating chytrid fungus in Costa Rica belongs to the virulent strain Bd-GPL- 2, which has been related to massive amphibian deaths worldwide; however, the isolates obtained showed genetic and morphological variation. Furthermore, we isolated epidermal bacteria from 12 amphibian species of surviving populations, some in danger of extinction, and evaluated their inhibitory activity against the collection of chytrid isolates. Through bioassays we confirmed the presence of chytrid-inhibitory bacterial genera in Costa Rican amphibians. However, we observed that the inhibition varied between different isolates of the same bacterial genus, and each bacterial isolation inhibited fungal isolation differently. In total, 14 bacterial isolates belonging to the genera Stenotrophomonas, Streptomyces, Enterobacter, Pseudomonas and Klebsiella showed inhibitory activity against all Bd isolates. Given the observed variation both in the pathogen and in the bacterial inhibition capacity, it is highly relevant to include local isolates and to consider the origin of the microorganisms when performing in vivo infection tests aimed at developing and implementing mitigation strategies for chytridiomycosis.United States Fish and Wildlife Service/[46-6003541]/USFWS/Estados UnidosUniversidad de Costa Rica/[801-B2-029]/UCR/Costa RicaUniversidad de Costa Rica/[810-B7-A46]/UCR/Costa RicaMinisterio de Ciencia, Tecnología y Telecomunicaciones/[849-PINN-2015-I]/MICITT/Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Estructuras Microscópicas (CIEMIC)UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM

Characterization of Batrachochytrium dendrobatidis Inhibiting Bacteria from Amphibian Populations in Costa Rica

Global amphibian declines and extinction events are occurring at an unprecedented rate. While several factors are responsible for declines and extinction, the fungal pathogen Batrachochytrium dendrobatidis (Bd) has been cited as a major constituent in these events. While the effects of this chytrid fungus have been shown to cause broad scale population declines and extinctions, certain individuals and relict populations have shown resistance. This resistance has been attributed in part to the cutaneous bacterial microbiome. Here, we present the first study characterizing anti-Bd bacterial isolates from amphibian populations in Costa Rica, including the characterization of two strains of Serratia marcescens presenting strong anti-Bd activity. Transcriptome sequencing was utilized for delineation of shifts in gene expression of the two previously uncharacterized strains of S. marcescens grown in three different treatments comprising Bd, heat-killed Bd, and a no Bd control. These results revealed up- and down-regulation of key genes associated with different metabolic and regulatory pathways. This information will be valuable in continued efforts to develop a bacterial-based approach for amphibian protection as well as providing direction for continued mechanistic inquiries of the bacterial anti-Bd response.Fish and Wildlife Service/[46-6003541]/USFWS/Estados UnidosUniversidad de Costa Rica/[801-B2-029]/UCR/Costa RicaGeneral Medicine Sciences of the National Institutes of Health/[P20GM103443]7NIGMS/Estados UnidosUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Estructuras Microscópicas (CIEMIC)UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM)UCR::Vicerrectoría de Docencia::Salud::Facultad de Medicina::Escuela de Medicin

The Arctic in the twenty-first century: changing biogeochemical linkages across a paraglacial landscape of Greenland

The Kangerlussuaq area of southwest Greenland encompasses diverse ecological, geomorphic, and climate gradients that function over a range of spatial and temporal scales. Ecosystems range from the microbial communities on the ice sheet and moisture-stressed terrestrial vegetation (and their associated herbivores) to freshwater and oligosaline lakes. These ecosystems are linked by a dynamic glacio-fluvial-aeolian geomorphic system that transports water, geological material, organic carbon and nutrients from the glacier surface to adjacent terrestrial and aquatic systems. This paraglacial system is now subject to substantial change because of rapid regional warming since 2000. Here, we describe changes in the eco- and geomorphic systems at a range of timescales and explore rapid future change in the links that integrate these systems. We highlight the importance of cross-system subsidies at the landscape scale and, importantly, how these might change in the near future as the Arctic is expected to continue to warm

Impact of clinical phenotypes on management and outcomes in European atrial fibrillation patients: a report from the ESC-EHRA EURObservational Research Programme in AF (EORP-AF) General Long-Term Registry

Background: Epidemiological studies in atrial fibrillation (AF) illustrate that clinical complexity increase the risk of major adverse outcomes. We aimed to describe European AF patients\u2019 clinical phenotypes and analyse the differential clinical course. Methods: We performed a hierarchical cluster analysis based on Ward\u2019s Method and Squared Euclidean Distance using 22 clinical binary variables, identifying the optimal number of clusters. We investigated differences in clinical management, use of healthcare resources and outcomes in a cohort of European AF patients from a Europe-wide observational registry. Results: A total of 9363 were available for this analysis. We identified three clusters: Cluster 1 (n = 3634; 38.8%) characterized by older patients and prevalent non-cardiac comorbidities; Cluster 2 (n = 2774; 29.6%) characterized by younger patients with low prevalence of comorbidities; Cluster 3 (n = 2955;31.6%) characterized by patients\u2019 prevalent cardiovascular risk factors/comorbidities. Over a mean follow-up of 22.5 months, Cluster 3 had the highest rate of cardiovascular events, all-cause death, and the composite outcome (combining the previous two) compared to Cluster 1 and Cluster 2 (all P <.001). An adjusted Cox regression showed that compared to Cluster 2, Cluster 3 (hazard ratio (HR) 2.87, 95% confidence interval (CI) 2.27\u20133.62; HR 3.42, 95%CI 2.72\u20134.31; HR 2.79, 95%CI 2.32\u20133.35), and Cluster 1 (HR 1.88, 95%CI 1.48\u20132.38; HR 2.50, 95%CI 1.98\u20133.15; HR 2.09, 95%CI 1.74\u20132.51) reported a higher risk for the three outcomes respectively. Conclusions: In European AF patients, three main clusters were identified, differentiated by differential presence of comorbidities. Both non-cardiac and cardiac comorbidities clusters were found to be associated with an increased risk of major adverse outcomes