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Wetlands cover a small portion of the world, but have disproportionate influence on global carbon (C) sequestration, carbon dioxide and methane emissions, and aquatic C fluxes. However, the underlying biogeochemical processes that affect wetland C pools and fluxes are complex and dynamic, making measurements of wetland C challenging. Over decades of research, many observational, experimental, and analytical approaches have been developed to understand and quantify pools and fluxes of wetland C. Sampling approaches range in their representation of wetland C from short to long timeframes and local to landscape spatial scales. This review summarizes common and cutting-edge methodological approaches for quantifying wetland C pools and fluxes. We first define each of the major C pools and fluxes and provide rationale for their importance to wetland C dynamics. For each approach, we clarify what component of wetland C is measured and its spatial and temporal representativeness and constraints. We describe practical considerations for each approach, such as where and when an approach is typically used, who can conduct the measurements (expertise, training requirements), and how approaches are conducted, including considerations on equipment complexity and costs. Finally, we review key covariates and ancillary measurements that enhance the interpretation of findings and facilitate model development. The protocols that we describe to measure soil, water, vegetation, and gases are also relevant for related disciplines such as ecology. Improved quality and consistency of data collection and reporting across studies will help reduce global uncertainties and develop management strategies to use wetlands as nature-based climate solutions., Supplementary Information: The online version contains supplementary material available at 10.1007/s13157-023-01722-2., Competing Interests: Competing InterestsThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© The Author(s) 2023.)

The entry of antibiotic resistance genes (ARGs) into aquatic systems has been documented for large municipal wastewater treatment plants (WWTPs), but there is less study of the impact of smaller plants that are situated on small rural rivers. We sampled water metagenomes for ARGs and taxa composition from the Kokosing River, a small rural river in Knox County, Ohio, which has been designated an Ohio State Scenic River for retention of natural character. Samples were obtained 1.0 km upstream, 120 m downstream, and 6.4 km downstream from the effluent release of the Mount Vernon WWTP. ARGs were identified in metagenomes using ShortBRED markers from the comprehensive antibiotic resistance database (CARD) screened against UniPROT. Through all seasons, the metagenome just downstream of the WWTP effluent showed a substantial elevation of at least 15 different ARGs, including 6 ARGs commonly associated with Acinetobacter baumannii, such as msrE , mphE (macrolide resistance), and tet (39) (tetracycline resistance). The ARGs most prevalent near the effluent pipe persisted 6.4 km downriver. Using metagenomic phylogenetic analysis (MetaPhlAn2) clade-specific marker genes, the taxa distribution near the effluent showed elevation of reads annotated as Acinetobacter species as well as gut-associated taxa, Bacteroides and Firmicutes . The ARG levels and taxa prevalence showed little dependence on seasonal chlorination of the effluent. Nitrogen and phosphorus were elevated near the effluent pipe but had no consistent correlation with ARG levels. We show that in a rural river microbiome, year-round wastewater effluent substantially elevates ARGs, including those associated with multidrug-resistant A. baumannii. IMPORTANCE Antibiotic resistance is a growing problem worldwide, with frequent transmission between pathogens and environmental organisms. Rural rivers can support high levels of recreational use by people unaware of inputs from treated wastewater, while wastewater treatment plants (WWTPs) can generate a small but significant portion of flow volume into a river surrounded by forest and agriculture. There is little information on the rural impacts of WWTP effluent on the delivery and transport of antibiotic resistance genes. In our study, the river water proximal to wastewater effluent shows evidence for the influx of multidrug-resistant Acinetobacter baumannii, an opportunistic pathogen of concern for hospitals but also widespread in natural environments. Our work highlights the importance of wastewater effluent in management of environmental antibiotic resistance, even in high quality, rural river systems.

In 2011, the US Environmental Protection Agency and its partners conducted the first National Wetland Condition Assessment at the continental-scale of the conterminous United States. A probability design for site selection was used to allow an unbiased assessment of wetland condition. We developed a vegetation multimetric index (VMMI) as a parsimonious biological indicator of ecological condition applicable to diverse wetland types at national and regional scales. Vegetation data (species presence and cover) were collected from 1138 sites that represented seven broad estuarine intertidal and inland wetland types. Using field collected data and plant species trait information, we developed 405 candidate metrics with potential for distinguishing least disturbed (reference) from most disturbed sites. Thirty-five of the metrics passed range, repeatability, and responsiveness screens and were considered as potential component metrics for the VMMI. A permutation approach was used to calculate thousands of randomly constructed potential national-scale VMMIs with 4, 6, 8, or 10 metrics. The best performing VMMI was identified based on limited redundancy among constituent metrics, sensitivity, repeatability, and precision. This final VMMI had four broadly applicable metrics (floristic quality index, relative importance of native species, richness of disturbance-tolerant species, and relative cover of native monocots). VMMI values and weights from the survey design for probability sites (n = 967) were used to estimate wetland area in good, fair, and poor condition, nationally and for each of 10 ecoregion by wetland type reporting groups. Strengths and limitations of the national VMMI for describing ecological condition are highlighted.

The use of models by ecologists and environmental managers, to inform environmental management and decision-making, has grown exponentially in the past 50 years. Due to logistical, economical, and theoretical benefits, model users frequently transfer preexisting models to new sites where data are scarce. Modelers have made significant progress in understanding how to improve model generalizability during model development. However, models are always imperfect representations of systems and are constrained by the contextual frameworks used during their development. Thus, model users need better ways to evaluate the possibility of unintentional misapplication when transferring models to new sites. We propose a method of describing a model's application niche for use during the model selection process. Using this method, model users synthesize information from databases, past studies, and/or past model transfers to create model performance curves and heat maps. We demonstrated this method using an empirical model developed to predict the ecological condition of plant communities in riverine wetlands of the Appalachian Highland physiographic region, U.S.A. We assessed this model's transferability and generalizability across (1) riverine wetlands in the contiguous U.S.A., (2) wetland types in the Appalachian Highland physiographic region, and (3) wetland types in the contiguous U.S.A. With this methodology and a discussion of its critical steps, we set the stage for further inquiries into the development of consistent and transparent practices for model selection when transferring a model.

Wetland soils contain some of the highest stores of soil carbon in the biosphere. However, there is little understanding of the quantity and distribution of carbon stored in our remaining wetlands or of the potential effects of human disturbance on these stocks. Here we use field data from the 2011 National Wetland Condition Assessment to provide unbiased estimates of soil carbon stocks for wetlands at regional and national scales. We find that wetlands in the conterminous United States store a total of 11.52 PgC, much of which is within soils deeper than 30 cm. Freshwater inland wetlands, in part due to their substantial areal extent, hold nearly ten-fold more carbon than tidal saltwater sites-indicating their importance in regional carbon storage. Our data suggest a possible relationship between carbon stocks and anthropogenic disturbance. These data highlight the need to protect wetlands to mitigate the risk of avoidable contributions to climate change.

We measured soil properties, carbon and nutrient (nitrogen, phosphorus) pools, ambient and potential denitrification, and phosphorus sorption index (PSI) in natural depressional wetlands and depressional wetlands restored through the U.S. Department of Agriculture (USDA) Wetland Reserve Program. We measured the same suite of variables in natural and USDA Conservation Reserve Program-restored riparian buffers and in agricultural fields adjacent to both systems to determine the degree to which ecosystem services are being provided through restoration in different hydrogeomorphic settings. Organic carbon and nutrient pools, PSI, and denitrification were greater in natural than in 5- to 10-yr-old restored depressional wetlands. In riparian soils, carbon and nutrient pools, PSI, and denitrification were comparable between restored and natural systems, suggesting that these services develop quickly after restoration. Restored depressional wetlands had lower soil organic C, N, and P relative to agricultural soils, whereas the opposite trend was observed in restored riparian soils. Four-year-old restored riparian buffers achieved equivalence to natural riparian buffers within 4 yr, whereas restored depressional wetlands took longer to provide these ecosystem services (i.e., PSI, denitrification, C storage) at levels comparable to natural wetlands. Restored depressional wetlands and riparian buffers provide ecosystem services lost through previous conversion to agriculture throughout the Midwest; however, the development of these services depends on hydrodynamics (pulsed versus nonpulsed), parent material, soil texture (sand, clay), and disturbance regime (prescribed fire) of the site. As restoration continues throughout the region, C sequestration and nutrient removal in these systems is expected to increase water quality at the local and regional levels., (Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.)

We sampled 92 wetlands from four different basins in the United States to quantify observer repeatability in rapid wetland condition assessment using the Delaware Rapid Assessment Protocol (DERAP). In the Inland Bays basin of Delaware, 58 wetland sites were sampled by multiple observers with varying levels of experience (novice to expert) following a thorough training workshop. In the Nanticoke (Delaware/Maryland), Cuyahoga (Ohio), and John Day (Oregon) basins, 34 wetlands were sampled by two expert teams of observers with minimal protocol training. The variance in observer to observer scoring at each site was used to calculate pooled standard deviations (SD(pool)), coefficients of variation, and signal-to-noise ratios for each survey. The results showed that the experience level of the observer had little impact on the repeatability of the final rapid assessment score. Training, however, had a large impact on observer to observer repeatability. The SD(pool) in the Inland Bay survey with training (2.2 points out of a 0-30 score) was about half that observed in the other three basins where observers had minimal training (SD(pool) = 4.2 points). Using the results from the survey with training, we would expect that two sites assessed by different, trained observers who obtain DERAP scores differing by more than 4 points are highly likely to differ in ecological condition, and that sites with scores that differ by 2 or fewer points are within variability that can be attributed to observer differences.