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Abstract Allosteric modulation is a central mechanism for metabolic regulation but has yet to be described for a gut microbiota-host interaction. Phenylacetylglutamine (PAGln), a gut microbiota-derived metabolite, has previously been clinically associated with and mechanistically linked to cardiovascular disease (CVD) and heart failure (HF). Here, using cells expressing β1- versus β2-adrenergic receptors (β1AR and β2AR), PAGln is shown to act as a negative allosteric modulator (NAM) of β2AR, but not β1AR. In functional studies, PAGln is further shown to promote NAM effects in both isolated male mouse cardiomyocytes and failing human heart left ventricle muscle (contracting trabeculae). Finally, using in silico docking studies coupled with site-directed mutagenesis and functional analyses, we identified sites on β2AR (residues E122 and V206) that when mutated still confer responsiveness to canonical β2AR agonists but no longer show PAGln-elicited NAM activity. The present studies reveal the gut microbiota-obligate metabolite PAGln as an endogenous NAM of a host GPCR.

Climate change is increasingly recognized as a serious threat to ecological systems, particularly freshwater systems. As a result, climate adaptation planning is more common in natural resource management. Because conservation resources are limited, decision support tools can help managers prioritize actions. We created the Species and Habitat-based Ecological Decision Support tool (wtrSHEDS) for conservation managers by integrating two distinct indices: one biologically based on aquatic watershed-scale species assemblages and the other a habitat climate change vulnerability index. To do this, we evaluated 60 crayfish and fish species in West Virginia, USA, using the NatureServe Climate Change Vulnerability Index under an A1B emissions scenario. The species vulnerability assessment results indicated that most species we assessed are vulnerable to climate change. Four species of fish and two species of crayfish had extremely high vulnerability due to their dependence on a specific hydrological cycle, sensitivity to barriers, and reproductive dependence on a thermal niche. Aggregating species at the watershed scale (8-digit hydrological unit code [HUC]) using an adapted weighted sum indicated which assemblages were the most and least vulnerable to climate change. Secondly, we created a unique habitat vulnerability index using the established Northeast Association of Fish and Wildlife Agencies habitat vulnerability index and an index of land cover vulnerability from the Forecast Scenario model’s land cover change projections under the A1B emissions scenario to incorporate potential synergistic impacts of land use. Land cover projections were reclassified based on the assumption that developed land would have a higher potential to interact with climate change than forested lands synergistically. We combined the two habitat indices at the watershed scale (HUC 8) and performed a Hot Spot Analysis to 1) combine the assemblage and habitat vulnerability indices and 2) identify watersheds of conservation priority. This assessment methodology illustrates a means of identifying priority conservation areas in West Virginia that utilize pre-existing data, can be modified to suit management needs, and, therefore, one that managers across regions can readily apply to their system.

Abstract Homeodomains (HDs) are the second largest class of DNA binding domains (DBDs) among eukaryotic sequence-specific transcription factors (TFs) and are the TF structural class with the largest number of disease-associated mutations in the Human Gene Mutation Database (HGMD). Despite numerous structural studies and large-scale analyses of HD DNA binding specificity, HD-DNA recognition is still not fully understood. Here, we analyze 92 human HD mutants, including disease-associated variants and variants of uncertain significance (VUS), for their effects on DNA binding activity. Many of the variants alter DNA binding affinity and/or specificity. Detailed biochemical analysis and structural modeling identifies 14 previously unknown specificity-determining positions, 5 of which do not contact DNA. The same missense substitution at analogous positions within different HDs often exhibits different effects on DNA binding activity. Variant effect prediction tools perform moderately well in distinguishing variants with altered DNA binding affinity, but poorly in identifying those with altered binding specificity. Our results highlight the need for biochemical assays of TF coding variants and prioritize dozens of variants for further investigations into their pathogenicity and the development of clinical diagnostics and precision therapies.

Simulating shallow groundwater (SGW) flow dynamics and stream–SGW interactions using numerical modeling tools is necessary to develop a mechanistic understanding of water flow systems and improve confidence in water resource management practices. A three-dimensional (3D) SGW flow model was developed for a riparian wetland in a mixed forest and agricultural catchment in West Virginia (WV), Appalachia, USA, using a Modular 3D Groundwater Model (MODFLOW). The MODFLOW simulation was calibrated in steady (R2 = 0.98, ME = −0.21, and RMSE = 0.77), transient state (R2 = 0.97, ME = −0.41, and RMSE = 1.28) and validated (R2 = 0.97, ME = −0.28, and RMSE = 1.05) using observed SGW levels from thirteen nested piezometers under steady and transient states. An experimental MT3D transport scenario was developed to show the lateral transport of NO₃-N from the aquifer to stream cells. Relatively stable SGW head distribution was observed. In the downstream reach, SGW discharge varied from 948 m3/day to 907 m3/day in 2020, with creek seepage ranging from 802 m3/day to 790 m3/day. Similarly, SGW input to the stream ranged from 891 m3/day to 978 m3/day, while creek seepage ranged from 796 m3/day to 800 m3/day in 2021. In upstream reaches, losing stream conditions were observed in January, June, and September 2020 and January to April 2021, while gaining stream conditions prevailed during other months. Thus, an approximately monthly alternating gaining–losing stream condition was observed in the upstream area. An experimental MT3D transport scenario resulted in an advection–dispersion scenario, showing a cumulative loss of 947 g of NO3-N from SGW to the stream. Denitrification accounted for the cumulative loss of 1406 g of NO3-N from SGW, surpassing 639 g of nitrate from the SGW to the stream during the study period. Additionally, particle tracking using MODPATH indicated a long residence time for SGW nutrients, affirming the efficiency of nitrogen transformation through denitrification. This study is among the first to simulate hydrologic and nutrient interactions in riparian wetlands of a mixed land use catchment in the Appalachian region of the northeastern United States. The results better inform water resource management decisions and modeling efforts in the Appalachian region and similar physiographic regions globally.

Abstract Understanding survival probabilities is critical for the sustainable harvest of wildlife and fisheries populations. Age‐ and stage class‐specific survival probabilities are needed to inform a suite of population models used to estimate abundance and track population trends. However, current techniques for estimating survival probabilities using age‐at‐harvest methods require restrictive assumptions or incorporate potentially unknown parameters within the model. Using a Bayesian approach, we developed a flexible age‐at‐harvest model that incorporates either age‐ or stage‐structured populations, while accounting for uncertainty in age structure, population growth rates and relative selectivity. Survival probabilities can vary by age or stage class, as well as by environmental covariates, and both population growth rates and selectivity for each age or stage class can be specified as fixed and known or these parameters can be specified as informative priors, allowing for the incorporation of expert opinion. We evaluated our model with simulations and empirical data from harvested bobcats Lynx rufus and American paddlefish Polyodon spathula. Models fit to simulated age‐at‐harvest data yielded unbiased estimates of survival probability when population growth rates and selectivity were centered on the data‐generating parameter. We obtained unbiased estimates of survival probability even with biased prior estimates of selectivity and random departures from the assumed stage distribution, although the latter increased uncertainty in those estimates. We found biased estimates of survival probability when the prior distribution for population growth rate was not centered on the data‐generating value. When fit to empirical harvest data, our proposed age‐at‐harvest model produced estimates of survival probability congruent to those reported in the literature within similar geographic regions. We demonstrate the utility of a novel age‐at‐harvest model that estimates survival probability and realistically account for uncertainty in model parameters, transcending the restrictive assumptions and auxiliary data requirements of other methods. Furthermore, we advise collecting information about population trends and age structure alongside age‐at‐harvest data to help reduce bias. Although our model cannot replace more rigorous methods, we believe our model will be transformative for wildlife and fisheries practitioners who collect age‐at‐harvest data to estimate age‐ or stage‐specific survival probabilities to help inform management decisions.

Precipitation patterns, water flow direction, and local land-use practices affect surface water (SW) and shallow groundwater (SGW) nutrient concentrations in riparian wetlands. Given physical process complexities, spatiotemporal quantification of nutrients and physical factors influencing nutrient concentrations are needed to advance wetland water resource management. To address these needs, a study was conducted in riparian wetlands of a mixed land-use catchment in West Virginia (WV), USA. Observed data included SW–SGW levels and nutrient concentrations, including nitrate (NO3-N), nitrite (NO2-N), ammonium (NH4-N), orthophosphate (PO43-P), total nitrogen (total_N), and total phosphorus (total_P) from January 2020 to December 2021. Water samples were collected monthly from stream gauge sites (n = 4) and co-located piezometers (n = 13). Results showed that, on average, gaining stream conditions were observed in upstream sites, and losing stream conditions were observed in downstream sites. Observed nutrient profiles between SW and SGW included SW exhibiting a higher average NO3-N concentration (0.42 mg/L), while SGW displayed an elevated NH4-N concentration (1.55 mg/L) relative to other nitrogen species. Significantly high (p < 0.05) SW NO3- concentrations in summer and fall were attributed to increased precipitation and corresponding water level and, therefore, pressure head and transport fluctuations. Principal Component Analysis (PCA) showed differences in nutrient concentrations based on the water source type and catchment land use, explaining 65% of data variability. Spearman correlation analysis illustrated the correlation among nutrient concentrations, land use, and water level changes in SW and SGW environments. This study provides needed baseline data on nutrient dynamics for a riparian wetland in a mixed land-use catchment, supplying science-based information to advance land and water management practices in the study watershed and similar physiographic watersheds globally.

Wetlands are important ecosystems that contribute to the sustainability of global ecosystems and provide ecosystem functions and services to human civilization. However, many anthropogenic land use practices have led to the degradation of wetlands, making them globally imperiled ecosystems. Within the United States, wetland mitigation is a federally regulated restoration strategy that offsets and compensates for impacts on aquatic resources through restoration. Performance standards assess post-restoration ecosystem development and help regulate management actions. The primary objective of this study is to investigate the organization and interactions of states and federal agencies in determining wetland mitigation performance standards. Using a mixed method approach, including semi-structured interviews and online database reviews, we identify decision-making drivers from the state agency perspective. We develop a ranking classification of state legislation that references performance standards and describes guidance documents by type of authorship. Our findings detail the results of our inquiry into each state’s procedures, including performance standards, revealing diverse management approaches across the nation as states play various implementation and regulatory roles and are driven by collaboration and negotiation among regulators, state and federal legislation, and guidance documents. In addition, we found performance standards most often assess biotic characteristics, with vegetative criteria being the most common. This study synthesizes performance-standard determination and criteria derived from interviews across a spectrum of federal and state participants and a series of guidance documents. We have built a database of these criteria by state and theme to improve our understanding of the dynamic interplay between wetland mitigation science, practice, and policy. Our findings are discussed in the context of the 2023 Sackett vs. United States Environmental Protection Agency ruling.

Summary: High-protein diets are promoted for individuals with type 2 diabetes (T2D). However, effects of dietary protein interventions on (gut-derived) metabolites in T2D remains understudied. We therefore performed a multi-center, randomized-controlled, isocaloric protein intervention with 151 participants following either 12-week high-protein (HP; 30Energy %, N = 78) vs. low-protein (LP; 10 Energy%, N = 73) diet. Primary objectives were dietary effects on glycemic control which were determined via glycemic excursions, continuous glucose monitors and HbA1c. Secondary objectives were impact of diet on gut microbiota composition and -derived metabolites which were determined by shotgun-metagenomics and mass spectrometry. Analyses were performed using delta changes adjusting for center, baseline, and kidney function when appropriate.This study found that a short-term 12-week isocaloric protein modulation does not affect glycemic parameters or weight in metformin-treated T2D. However, the HP diet slightly worsened kidney function, increased alpha-diversity, and production of potentially harmful microbiota-dependent metabolites, which may affect host metabolism upon prolonged exposure.

Macrophytes have the potential to withstand pollutant-induced stress and can be used to clean contaminated water using phyto-extraction, phyto-degradation, phyto-filtration, phyto-stimulation, and phyto-volatilization technique(s). Phytoremediation through constructed wetlands (CWs) for eliminating inorganic and organic pollutants from household sewage and wastewater has attracted scientific attention. CWs are artificially engineered treatment systems that utilize natural cycles or processes involving soils, wetland vegetation, and plant and soil-associated microbial assemblages to remediate contaminated water and improve its quality. Herein, we present a detailed assessment of contaminant removal effectiveness in different CW systems, i.e., free-water surface or surface-flow constructed wetlands (FWSCWs/SFCWs), subsurface-flow constructed wetlands (SSFCWs), and hybrid constructed wetlands (HCWs). Several wetland floral species have been reported as potential phytoremediators, effectively reducing aquatic contamination through biodegrading, biotransforming, and bioaccumulating contaminants. Water hyacinth (Pontederia crassipes) is one of the most resistant macrophytes, capable of tolerating high nitrate (NO3−) and phosphate (PO42−) concentrations. Other aquatic weeds also effectively alleviate biological oxygen demand (BOD), chemical oxygen demand (COD), total dissolved solids (TDS), and pathogen levels and ameliorate the impact of different ionic forms of nitrogen (N), phosphorus (P), and trace elements (TEs). The review primarily focuses on using hydrophyte(s)-microbe(s) associations in different CWs as an essential phytoremediation tool for sustainable management of freshwater ecosystems, ecorestoration, and prospective resource recovery, favoring a circular bioeconomy (CBE).

Abstract Anthropogenic land use changes have broad impacts on biological diversity, often resulting in shifts in community composition. While many studies have documented negative impacts on occurrence and abundance of species, less attention has been given to native species that potentially benefit from anthropogenic land use changes. For many species reaching high densities in human‐dominated landscapes, it is unclear whether these environments represent higher quality habitat than more natural environments. We examined the influence of landscape ecological integrity on relative abundance and body condition of two native generalist freshwater turtle species that are prevalent in anthropogenic systems, the painted turtle (Chrysemys picta) and red‐eared slider (Trachemys scripta elegans). Relative abundance was negatively associated with ecological integrity for both species, but the relationship was not strongly supported for painted turtles. Body condition was positively associated with ecological integrity for painted turtles, with no strong association for red‐eared sliders. Our study suggests that both species benefitted at the population level from reduced ecological integrity, but individual‐level habitat quality was reduced for painted turtles. The differing responses between these two habitat generalists could partially explain why red‐eared sliders have become a widespread exotic invasive species, while painted turtles have not.

The Ghodaghodi Lake Complex is a Ramsar site, Nepal’s first bird sanctuary, and has significant ecological and economic values. The lake complex is in the western part of the lowland of the Terai region. Numerous studies indicate a relation between the normalized difference vegetation index (NDVI), land use, and land cover with plant diversity. However, the association between terrestrial plant diversity and NDVI in the Ghodaghodi Lake Complex is unknown but has important implications due to potential land use changes. We aimed to understand the relationship between plant diversity and NDVI in the Ghodaghodi Lake Complex. We performed a vegetation survey using a simple random sampling methodology. Shannon–Wiener’s diversity index (H’) was calculated from the field data, and Landsat images were used to compare land use and land cover changes and calculate NDVI values for 2000 and 2022. The image classification shows that forest cover in April and December 2000 was 71.1% and 58.5%, respectively, and was the dominant land cover in the study area. In contrast, agriculture occupied 18.8% and 27.3% in April and December 2000, respectively, and was the primary land use. Forests covered the most land in April (64.8%) and December (65.3%) of 2022. Likewise, agriculture was a widespread land use. We found a significant correlation (r = 0.80, p < 0.05) between the NDVI and plant species diversity, as the NDVI explained 65% of plant species diversity. There was a decrease in forest cover from 2000 to 2022. The strong correlation between the NDVI and vegetation species diversity shows that the NDVI can be a substitute for plant diversity. Our findings show that increased NDVI corresponds to increased plant species diversity and that the lake complex had more plant diversity in 2022 than in 2000, despite a decrease in forested lands.

Abstract Acoustic communication of American Crocodiles (Crocodylus acutus) is relatively understudied. Our overall aim was to determine the acoustic structure of wild American Crocodile distress calls, distinguish call differences among size classes (hatchling, juvenile, sub‐adult, and adult), and investigate call production on a gradient of human disturbance. American Crocodile distress calls have strong frequency modulation and are comprised of multiple harmonics in a downsweeping pattern. Measured parameters (total duration, first quartile duration, maximal frequency, first quartile frequency, end frequency, slope of first quartile, slope of last quartiles) differed significantly among size classes (p

Abstract In West Virginia, USA, there are 24 conservation easement program wetlands enrolled in the Agricultural Conservation Easement Program (ACEP). These wetlands are located on private agricultural land and are passively managed. Due to their location within fragmented agricultural areas, wetlands enrolled in ACEP in West Virginia have the potential to add wetland ecosystem services in areas that are lacking these features. We evaluated ACEP wetlands compared to reference wetlands on public land in West Virginia by using surrounding land cover, vegetative cover, and wetland features and stressors such as the presence or absence of erosion, upland inclusion, algal mats, and evidence of impacts from the surrounding landscape as surrogate measurements of wetland function on 13 ACEP wetlands and 10 reference wetlands. ACEP wetlands had higher percentages of tree coverage and a higher proportion of agricultural land in the areas immediately surrounding the wetland. Reference wetlands had higher percent coverage of emergent vegetation and had a higher proportion of forest in the immediate landscape. Our findings suggest that ACEP wetlands provide valuable early successional and forested wetland cover in a state that is largely forested. Because of this, it is important to maintain and even expand ACEP in West Virginia to continue providing a valuable source of early successional wetland habitat.

Abstract Over the past 20 years, the use of non‐invasive hair snare surveys in wildlife research and management has become more prevalent. While these tools have been used to answer important research questions, these techniques often fail to gather information on elusive carnivores, such as bobcats (Lynx rufus). Due to the limited success of previous bobcat studies using hair snares which required active rubbing, this technique has largely fallen out of use, in favor of camera trapping. The goal of our study was to construct a novel, passive bobcat hair snare that could be deployed regardless of terrain or vegetation features, which would be effective for use in capture–recapture population estimation at a large spatial scale. This new hair snare was deployed in 1500 10‐km2 cells across West Virginia (USA) between two sampling seasons (2015–2016). Collected hair samples were analyzed with newly developed mitochondrial DNA primers specifically for felids and qPCR to determine species of origin, with enough sensitivity to identify samples as small as two bobcat hairs. Over the two years of the study, a total of 378 bobcat detections were recorded from 42,000 trap nights of sampling, for an overall rate of 0.9 detections/100 trap nights—nearly 2–6 times greater than any previous bobcat hair snare study. While the overall number of recaptured animals was low (n = 9), continued development of this platform should increase its usefulness in capture–recapture studies.

Population genetics can reveal whether colonization of created habitats has been successful and inform future strategies for habitat creation. We used genetic analysis to investigate spotted salamander (Ambystoma maculatum) colonization of created vernal pools and explored the impact of habitat characteristics on the genetic diversity and connectivity of the pools. Our first objective was to examine genetic structure, differentiation, diversity, and potential for a founder effect. Our second objective was to determine if habitat characteristics were associated with effective number of breeders, relatedness, or genetic diversity. We sampled spotted salamander larvae in 31 created vernal pools (1–5 years old) in Monongahela National Forest (WV) in May and June 2015 and 2016. The youngest pools exhibited genetic differentiation, a founder effect, and low effective number of breeders. Effective number of breeders was positively associated with pool age, vegetation cover, pool diameter, and sample size. Vegetation cover was also negatively associated with relatedness. Genetic diversity did not have strong environmental predictors. Our results indicated the effective number of breeders increased and genetic differentiation decreased within 4–5 years of pool creation, a sign of rapid colonization and potential population establishment. Our research also showed that higher vegetative cover within the pool and larger pool diameters could impact habitat quality and should be incorporated into future pool creation.

Abstract Water‐borne hormone measurement is a noninvasive method suitable for amphibians of all sizes that are otherwise difficult to sample. For this method, containment‐water is assayed for hormones released by the animal. Originally developed in fish, the method has expanded to amphibians, but requires additional species‐specific validations. We wanted to determine physiological relevance of water‐borne corticosterone in spotted salamanders (Ambystoma maculatum) by comparing concentrations to those taken using established corticosterone sampling methods, such as plasma. Using a mixture of field and laboratory studies, we compared water‐borne corticosterone levels to other traditional methods of sampling corticosterone for spotted salamander larvae, metamorphs, and adults. Despite multiple attempts, and detecting differences between age groups, we found no correlations between water‐borne and plasma corticosterone levels in any age group. Water‐borne sampling measures a rate of release; whereas plasma is the concentration circulating in the blood. The unique units of measurement may inherently prevent correlations between the two. These two methods may also require different interpretations of the data and the physiological meaning. We also note caveats with the method, including how to account for differences in body size and life history stages. Collectively, our results illustrate the importance of careful validation of water‐borne hormone levels in each species in order to understand its physiological significance.

Wetland indices of biological integrity (IBIs) are a common component in monitoring the wetland water resources as required by the United States’ Clean Water Act (CWA). The effectiveness of an IBI to monitor disturbance is dependent on the metrics being consistently responsive to measures of human disturbance within a described classification category. We present IBIs designed for two types of commonly used wetland classification systems – the hydrogeomorphic (HGM) and the National Wetlands Inventory (NWI). The metrics making up the IBIs were derived from anuran, avian, macroinvertebrate, and vegetation communities; each representing increasing levels of resources associated with gathering the necessary data. Knowing which communities’ data best corresponds to impairment can maximize limited wetland monitoring resources, especially if the response differs based on the wetland vegetation type NWI or HGM position on the landscape. By combining these two classification schemes together, to better define a wetland’s form and context on the landscape, more of the variability in community metrics are explained by human impairment. Moreover, when multiple taxa are used within a single wetland classification scheme, the response of the multi-taxa community IBI to the human disturbance gradient is often more sensitive than one-taxa group alone. This approach, a combination of taxa, in hybrid 2-system classification schemes, creates additional utility in measuring the effectiveness of wetland assessments and, or restoration success.

Wetland restoration is a common practice, and, in many cases, it is for mitigation to offset losses of natural wetlands due to human interference. Researchers commonly compare bird, amphibian, and reptile communities between these wetlands and natural wetlands but overlook small mammals. However, terrestrial small mammals are essential to consider as they serve a fundamental role in the ecosystem as seed dispersers and prey for larger wildlife. We conducted small mammal trapping on 26 wetlands (n = 14 restored, n = 12 natural) in West Virginia, USA, in the summers of 2020 and 2021 to obtain and compare community metrics between wetland types. We found that mass, occupancy probability, and community composition were similar between restored and natural wetlands. However, the apparent abundance of deer mice (Peromyscus maniculatus) was higher in natural wetlands (p < 0.001). Because we captured the three rarest species exclusively in natural wetlands, the ability of restored wetlands to provide an adequate habitat for rare or wetland-obligate species may be biologically significant. Restored wetlands mainly offer sufficient habitat for small mammal communities, but apparent abundance in restored wetlands may differ from natural wetlands depending on species.

The main factors controlling plant litter decomposition rates are litter quality and environmental factors. We investigated how different salinity and inundation conditions influence the decomposition rate and how litter quality affects dynamic change during the decomposition process in the Yellow River Delta wetland in northeastern China. To do this, we designed a field experiment using the litter bag method to study two selected dominant halophyte species (Phragmites australis and Suaeda salsa). We found litter decomposed faster when inundated across a range of salinities. Both the water chemical oxygen demand and NH3-N concentration were inversely correlated with salinity. The ratio of nitrogen to phosphorous showed an upward trend during the process of decomposition. The decomposition rate increased during the later stage of the process. Under inundation conditions, the decomposition rate of only S. salsa showed a positive correlation with salinity. The study results suggest that moderately higher salinity would increase anaerobic decomposition owing to the combined effect of salinity and inundation. The implications of our findings may be used to further assess the impact of environmental parameters and litter quality on the decomposition rate in estuarine wetland and can help determine a strategy for wetland reparation and remediation.

Wetland mitigation efforts have increased in numbers over the past two decades to combat wetland loss in the United States. Data regarding wetland function such as biodiversity are required to be collected 5–10 years after a project is complete; however, pre-restoration data that can inform the effectiveness of mitigation are often not collected. We conducted pre-restoration surveys on various taxa along or within Ruby Run, a tributary of Deckers Creek in north-central West Virginia, USA, from 2016 to 2020 to determine the baseline relative abundance and diversity within the stream and the associated riparian zone. In five years, we observed 237 species (154 plant, 58 bird, 13 fish, 6 small mammal, and 6 anuran) and 25 families of macroinvertebrates. Seasonal fluctuations in diversity were present, but mean diversity was relatively consistent among years across taxa, except in anurans, where there was a decrease each year. Wetland mitigation efforts should continue to be monitored for success using multiple taxa, because land use change can affect taxa in different ways, resulting in well-rounded assessments that can improve wetland management practices.

Research and field monitoring can disturb wetland integrity. Adoption of ethical field practices is needed to limit monitoring induced stressors such as trampling, non-native seed and invertebrate dispersal, and disease and fungal spread. We identify a linear pathway of deterioration highlighting stressors that can progress to cumulative impacts, consequences, and losses at the site scale. The first step to minimize disturbance is to assess and classify the current ecosystem quality. We present a tiered framework for wetland classification and link preventative measures to the wetland tier. Preventative measures are recommended at various intensities respective to the wetland tier, with higher tiered wetlands requiring more intense preventative measures. In addition, preventative measures vary by time of implementation (before, during, and after the wetland visit) to mitigate impacts at various temporal scales. The framework is designed to increase transparency of field monitoring impacts and to promote the adoption of preventative measures. Implementing preventative measures can build accountability and foster a greater appreciation for our roles as researchers and managers in protecting wetlands.

Habitat loss and degradation have contributed to declining populations of stiff-tailed ducks including the migratory northern ruddy duck (Oxyura jamaicensis rubida), and non-migratory masked duck (Nomonyx dominicus) and West Indian ruddy duck (Oxyura j. jamaicensis). Studies collecting time-activity budgets on waterfowl can provide important insight into habitat use, behavior, and niche partitioning. Even though the northern ruddy duck and West Indian ruddy duck are recognized as the same species, we treated them separately to appraise possible ecological differences. We recorded 24-h time-activity budgets for masked ducks (n = 142), northern ruddy ducks (n = 1,401), and West Indian ruddy ducks (n = 3,765) on the Laguna Cartagena National Wildlife Refuge in Lajas, Puerto Rico from January through April 2015 and 2016. Behaviors varied among taxa, between sexes, and between diurnal and nocturnal sampling periods. Resting and sleeping were common behaviors observed during the day and night, but all 3 taxa fed more at night with masked duck feeding more than the West Indian ruddy duck and northern ruddy duck. Northern ruddy duck and West Indian ruddy duck time-activity budgets varied with 7 of 8 behavioral categories during the day and 5 of 8 behavioral categories at night differing. Ecological differences such as body size and migration of northern ruddy ducks may account for the differences in behavior when compared to non-migratory West Indian ruddy ducks. Keywords: Laguna Cartagena National Wildlife refuge, Masked duck, Northern ruddy duck, Time-activity budgets, Waterfowl behavior, West Indian ruddy duck

Canaan Valley, West Virginia, USA provides habitat for many plants considered rare in West Virginia. The local white-tailed deer (Odocoileus virginianus Zimm.) are a popular attraction for visitors as well as resident and nonresident hunters. However, concerns exist over the impact of white-tailed deer herbivory on rare plant communities in the wetlands. We evaluated stakeholder attitudes regarding white-tailed deer management and rare plant conservation by mailing surveys to property owners in Canaan Valley and Tucker County, WV, and providing surveys to visitors at Canaan Valley State Park and National Wildlife Refuge. Individuals that had incurred crop damage were more likely to view the Canaan Valley population as overabundant while nonresident landowners placed higher values on the conservation of rare plants and plant communities in Canaan Valley. We found that hunters were more likely than nonhunters to support white-tailed deer management actions. Results of this survey may help managers to direct management and education goals toward actual rather than perceived stakeholder attitudes. Keywords: White-tailed deer, Stakeholder, Nonresident, Visitor, Plant conservation

There is no reliable method for determining age for most species of long-lived birds. Recent success using the skin chemical pentosidine as a biomarker has shown promise as an aging tool for birds. Pentosidine levels have been determined only from the breast tissue of carcasses, and we sought to refine the procedure with respect to biopsy size and location for safe and effective use on living birds. We compared pentosidine concentrations in 4 skin-size samples (4, 6, 8, and 20-mm diameter biopsies) from the breast of black vulture (Coragyps atratus) carcasses. We also compared pentosidine levels from breast and patagial tissue to document potential differences among collection sites of deceased vultures (with unknown ages) and monk parakeets (Myiopsitta monachus; with actual, minimal, and unknown ages). Pentosidine concentrations (pmol pentosidine/mg collagen) were similar among the 4 sizes of vulture breast skin (P = 0.82). Pentosidine concentrations for the breast (x̄ = 8.9, SE = 0.55, n = 28) and patagium (x̄ = 8.9, SE = 0.51, n = 28) of vultures were similar, but in parakeets, pentosidine was higher in the breast (x̄ = 15.9, SE = 1.30, n = 105) than the patagium (x̄ = 11.5, SE = 1.10, n = 105). We made pentosidine-based age estimates for vultures and parakeets using a general age curve for wild birds. We also made vulture age estimates using plumage characteristics and a cormorant (Phalacrocorax auritus) age curve. Vulture pentosidine-based age estimates appear to correspond to plumage-based age estimates. Pentosidine-based age estimates for 88% of the known-aged parakeets (n = 17) were within 6 months of actual ages. Even though known ages were not available for all birds, we found a positive trend in pentosidine versus age for both species. We suggest that 6-mm diameter skin samples from the patagium of living vultures and other similar-sized birds will provide sufficient tissue for reliable age estimation and will not impair flight ability.

Information on nonbreeding waterbirds using created wetlands in the Central Appalachian region of the United States is limited. We compared waterbird communities of two managed wetlands, created in 2013 and 2001, in West Virginia. We observed 27 species of waterbirds. Species richness and diversity were generally similar between the wetlands, but species composition and use differed. Branta canadensis (Canada Geese), Anas strepera (Gadwall), Bucephala albeola (Buffleheads), Aythya affinis (Lesser Scaup), and Aythya collaris (Ring-Necked Ducks) used the older wetland most frequently. Disparities in species use were the highest in March. The older wetland differed from the younger in supporting species such as diving ducks, possibly due to differences in size, vegetation, water depth, and microtopography. However, the ability to provide habitat for waterbirds during the winter was determined to be comparable between wetlands, despite their age difference.

Salamanders and riparian forests are intimately interconnected. Salamanders are integral to ecosystem functions, contributing to vertebrate biomass and complex food webs in riparian forests. In turn, these forests are critical ecosystems that perform many environmental services, facilitate high biodiversity and species richness, and provide habitat to salamander populations. Due to the global decline of amphibians, it is important to understand, as thoroughly and holistically as possible, the roles of environmental parameters and the impact of human activities on salamander abundance and diversity in riparian forests. To determine the population responses of salamanders to a variety of environmental factors and anthropogenic activities, we conducted a review of published literature that compared salamander abundance and diversity, and then summarized and synthesized the data into general patterns. We identify stream quality, leaf litter and woody debris, riparian buffer width, and soil characteristics as major environmental factors influencing salamander populations in riparian forests, describe and explain salamander responses to those factors, and discuss the effects of anthropogenic activities such as timber harvest, prescribed fires, urbanization, road construction, and habitat fragmentation. This review can assist land and natural resource managers in anticipating the consequences of human activities and preparing strategic conservation plans.

Natural stream channel design principles and riparian restoration practices were applied during spring 2010 to an agriculturally impaired reach of the Cacapon River, a tributary of the Potomac River which flows into the Chesapeake Bay. Aquatic macroinvertebrates and fishes were sampled from the restoration reach, two degraded control, and two natural reference reaches prior to, concurrently with, and following restoration (2009 through 2010). Collector filterers and scrapers replaced collector gatherers as the dominant macroinvertebrate functional feeding groups in the restoration reach. Before restoration, based on indices of biotic integrity (IBI), the restoration reach fish and macroinvertebrate communities closely resembled those sampled from the control reaches, and after restoration more closely resembled those from the reference reaches. Although the macroinvertebrate community responded more favorably than the fish community, both communities recovered quickly from the temporary impairment caused by the disturbance of restoration procedures and suggest rapid improvement in local ecological conditions.