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Building translational ecology communities of practice: insights from the field

Translational ecology (TE) prioritizes the understanding of social systems and decision contexts in order to address complex natural resource management issues. Although many practitioners in applied fields employ translational tactics, the body of literature addressing such approaches is limited. We present several case studies illustrating the principles of TE and the diversity of its applications. We anticipate that these examples will help others develop scientific products that decision makers can use “off the shelf” when solving critical ecological and social challenges. Our collective experience suggests that research of such immediate utility is rare. Long‐term commitment to working directly with partners to develop and reach shared goals is central to successful translation. The examples discussed here highlight the benefits of translational processes, including actionable scientific results, more informed policy making, increased investment in science‐driven solutions, and inspiration for partnerships. We aim to facilitate future TE‐based projects and build momentum for growing this community of practice.

Frontiers in Ecology and the Environment↗

Can’t see the random forest for the decision trees: Selecting predictive models for restoration ecology

Improving predictions of restoration outcomes is increasingly important to resource managers for accountability and adaptive management, yet there is limited guidance for selecting a predictive model from the multitude available. The goal of this paper was to identify an optimal predictive framework for restoration ecology using eleven modeling frameworks (including, machine learning, inferential, and ensemble approaches), and three data groups (field data, geographic data [GIS], and a combination thereof). We test this approach with a dataset from a large post-fire sagebrush reestablishment project in the Great Basin, USA. Predictive power varied among models and data groups, ranging from 58-79% accuracy. Finer scale field data generally had the greatest predictive power, although GIS data were present in the best models overall. An ensemble prediction computed from the ten models parameterized to field data was well above average for accuracy but was outperformed by others that prioritized model parsimony by selecting predictor variables based on rankings of their importance among all candidate models. The variation in predictive power among a suite of modeling frameworks underscores the importance of a model comparison and refinement approach that evaluates multiple models and data groups, and selects variables based on their contribution to predictive power. The enhanced understanding of factors influencing restoration outcomes accomplished by this framework has the potential to aid the adaptive management process for improving future restoration outcomes.

Restoration Ecology↗

Florida Integrated Science Center (FISC) Coral Reef Research

Coral reefs provide important ecosystem services such as shoreline protection and the support of lucrative industries including fisheries and tourism. Such ecosystem services are being compromised as reefs decline due to coral disease, climate change, overfishing, and pollution. There is a need for focused, integrated science to understand the complex ecological interactions and effects of these many stressors and to provide information that will effectively guide policies and best management practices to preserve and restore these important resources. The U.S. Geological Survey Florida Integrated Science Center (USGS-FISC) is conducting a coordinated Coral Reef Research Project beginning in 2009. Specific research topics are aimed at addressing priorities identified in the 'Strategic Science for Coral Ecosystems 2007-2011' document (U.S. Geological Survey, 2007). Planned research will include a blend of historical, monitoring, and process studies aimed at improving our understanding of the development, current status and function, and likely future changes in coral ecosystems. Topics such as habitat characterization and distribution, coral disease, and trends in biogenic calcification are major themes of understanding reef structure, ecological integrity, and responses to global change.

Fact Sheet↗

A method to detect discontinuities in census data

The distribution of pattern across scales has predictive power in the analysis of complex systems. Discontinuity approaches remain a fruitful avenue of research in the quest for quantitative measures of resilience because discontinuity analysis provides an objective means of identifying scales in complex systems and facilitates delineation of hierarchical patterns in processes, structure, and resources. However, current discontinuity methods have been considered too subjective, too complicated and opaque, or have become computationally obsolete; given the ubiquity of discontinuities in ecological and other complex systems, a simple and transparent method for detection is needed. In this study, we present a method to detect discontinuities in census data based on resampling of a neutral model and provide the R code used to run the analyses. This method has the potential for advancing basic and applied ecological research.

Ecology and Evolution↗

Can personality predict individual differences in brook trout spatial learning ability?

While differences in individual personality are common in animal populations, understanding the ecological significance of variation has not yet been resolved. Evidence suggests that personality may influence learning and memory; a finding that could improve our understanding of the evolutionary processes that produce and maintain intraspecific behavioural heterogeneity. Here, we tested whether boldness, the most studied personality trait in fish, could predict learning ability in brook trout. After quantifying boldness, fish were trained to find a hidden food patch in a maze environment. Stable landmark cues were provided to indicate the location of food and, at the conclusion of training, cues were rearranged to test for learning. There was a negative relationship between boldness and learning as shy fish were increasingly more successful at navigating the maze and locating food during training trials compared to bold fish. In the altered testing environment, only shy fish continued using cues to search for food. Overall, the learning rate of bold fish was found to be lower than that of shy fish for several metrics suggesting that personality could have widespread effects on behaviour. Because learning can increase plasticity to environmental change, these results have significant implications for fish conservation.

Behavioural Processes↗

West Florida shelf circulation and temperature budget for the 1999 spring transition

Mid-latitude continental shelves undergo a spring transition as the net surface heat flux changes from cooling to warming. Using in situ data and a numerical circulation model we investigate the circulation and temperature budget on the West Florida Continental Shelf (WFS) for the spring transition of 1999. The model is a regional adaptation of the primitive equation, Princeton Ocean Model forced by NCEP reanalysis wind and heat flux fields and by river inflows. Based on agreements between the modeled and observed fields we use the model to draw inferences on how the surface momentum and heat fluxes affect the seasonal and synoptic scale variability. We account for a strong southeastward current at mid-shelf by the baroclinic response to combined wind and buoyancy forcing, and we show how this local forcing leads to annually occurring cold and low salinity tongues. Through term-by-term analyses of the temperature budget we describe the WFS temperature evolution in spring. Heat flux largely controls the seasonal transition, whereas ocean circulation largely controls the synoptic scale variability. These two processes, however, are closely linked. Bottom topography and coastline geometry are important in generating regions of convergence and divergence. Rivers contribute to the local hydrography and are important ecologically. Along with upwelling, river inflows facilitate frontal aggregation of nutrients and the spring formation of a high concentration chlorophyll plume near the shelf break (the so-called ‘Green River’) coinciding with the cold, low salinity tongues. These features originate by local, shelf-wide forcing; the Loop Current is not an essential ingredient.

Gulf of Mexico↗

Activity-based, genome-resolved metagenomics uncovers key populations and pathways involved in subsurface conversions of coal to methane

Microbial metabolisms and interactions that facilitate subsurface conversions of recalcitrant carbon to methane are poorly understood. We deployed an in situ enrichment device in a subsurface coal seam in the Powder River Basin (PRB), USA, and used BONCAT-FACS-Metagenomics to identify translationally active populations involved in methane generation from a variety of coal-derived aromatic hydrocarbons. From the active fraction, high-quality metagenome-assembled genomes (MAGs) were recovered for the acetoclastic methanogen, Methanothrix paradoxum , and a novel member of the Chlorobi with the potential to generate acetate via the Pta-Ack pathway. Members of the Bacteroides and Geobacter also encoded Pta-Ack and together, all four populations had the putative ability to degrade ethylbenzene, phenylphosphate, phenylethanol, toluene, xylene, and phenol. Metabolic reconstructions, gene analyses, and environmental parameters also indicated that redox fluctuations likely promote facultative energy metabolisms in the coal seam. The active “ Chlorobi PRB” MAG encoded enzymes for fermentation, nitrate reduction, and multiple oxygenases with varying binding affinities for oxygen. “ M. paradoxum PRB” encoded an extradiol dioxygenase for aerobic phenylacetate degradation, which was also present in previously published Methanothrix genomes. These observations outline underlying processes for bio-methane from subbituminous coal by translationally active populations and demonstrate activity-based metagenomics as a powerful strategy in next generation physiology to understand ecologically relevant microbial populations.

Montana, Wyoming↗

Evaluating a multispecies adaptive management framework: Must uncertainty impede effective decision-making?

Application of adaptive management to complex natural resource systems requires careful evaluation to ensure that the process leads to improved decision-making. As part of that evaluation, adaptive policies can be compared with alternative nonadaptive management scenarios. Also, the value of reducing structural (ecological) uncertainty to achieving management objectives can be quantified. A multispecies adaptive management framework was recently adopted by the Atlantic States Marine Fisheries Commission for sustainable harvest of Delaware Bay horseshoe crabs Limulus polyphemus, while maintaining adequate stopover habitat for migrating red knots Calidris canutus rufa , the focal shorebird species. The predictive model set encompassed the structural uncertainty in the relationships between horseshoe crab spawning, red knot weight gain and red knot vital rates. Stochastic dynamic programming was used to generate a state-dependent strategy for harvest decisions given that uncertainty. In this paper, we employed a management strategy evaluation approach to evaluate the performance of this adaptive management framework. Active adaptive management was used by including model weights as state variables in the optimization and reducing structural uncertainty by model weight updating. We found that the value of information for reducing structural uncertainty is expected to be low, because the uncertainty does not appear to impede effective management. Harvest policy responded to abundance levels of both species regardless of uncertainty in the specific relationship that generated those abundances. Thus, the expected horseshoe crab harvest and red knot abundance were similar when the population generating model was uncertain or known, and harvest policy was robust to structural uncertainty as specified. Synthesis and applications . The combination of management strategy evaluation with state-dependent strategies from stochastic dynamic programming was an informative approach to evaluate adaptive management performance and value of learning. Although natural resource decisions are characterized by uncertainty, not all uncertainty will cause decisions to be altered substantially, as we found in this case. It is important to incorporate uncertainty into the decision framing and evaluate the effect of reducing that uncertainty on achieving the desired outcomes

Journal of Applied Ecology↗

Patterns and controls of foliar nutrient stoichiometry and flexibility across United States forests

Plant element stoichiometry and stoichiometric flexibility strongly regulate ecosystem responses to global change. Here, we tested three potential mechanistic drivers (climate, soil nutrients, and plant taxonomy) of both using paired foliar and soil nutrient data from terrestrial forested National Ecological Observatory Network sites across the USA. We found that broad patterns of foliar nitrogen (N) and foliar phosphorus (P) are explained by different mechanisms. Plant taxonomy was an important control over all foliar nutrient stoichiometries and concentrations, especially foliar N, which was dominantly related to taxonomy and did not vary across climate or soil gradients. Despite a lack of site-level correlations between N and environment variables, foliar N exhibited intraspecific flexibility, with numerous species-specific correlations between foliar N and various environmental factors, demonstrating the variable spatial and temporal scales on which foliar chemistry and stoichiometric flexibility can manifest. In addition to plant taxonomy, foliar P and N:P ratios were also linked to soil nutrient status (extractable P) and climate, especially actual evapotranspiration rates. Our findings highlight the myriad factors that influence foliar chemistry and show that broad patterns cannot be explained by a single consistent mechanism. Furthermore, differing controls over foliar N versus P suggests that each may be sensitive to global change drivers on distinct spatial and temporal scales, potentially resulting in altered ecosystem N:P ratios that have implications for processes ranging from productivity to carbon sequestration.

Ecology↗

Recurrent hybridization and recent origin obscure phylogenetic relationships within the ‘white-headed’ gull (Larus sp.) complex

Species complexes that have undergone recent radiations are often characterized by extensive allele sharing due to recent ancestry and (or) introgressive hybridization. This can result in discordant evolutionary histories of genes and heterogeneous genomes, making delineating species limits difficult. Here we examine the phylogenetic relationships among a complex group of birds, the white-headed gulls (Aves: Laridae), which offer a unique window into the speciation process due to their recent evolutionary history and propensity to hybridize. Relationships were examined among 17 species (61 populations) using a multilocus approach, including mitochondrial and nuclear intron DNA sequences and microsatellite genotype information. Analyses of microsatellite and intron data resulted in some species-based groupings, although most species were not represented by a single cluster. Considerable allele and haplotype sharing among white-headed gull species was observed; no locus contained a species-specific clade. Despite this, our multilocus approach provided better resolution among some species than previous studies. Interestingly, most clades appear to correspond to geographic locality: our BEAST analysis recovered strong support for a northern European/Icelandic clade, a southern European/Russian clade, and a western North American/ canus clade, with weak evidence for a high latitude clade spanning North America and northwestern Europe. This geographical structuring is concordant with behavioral observations of pervasive hybridization in areas of secondary contact. The extent of allele and haplotype sharing indicates that ecological and sexual selection are likely not strong enough to complete reproductive isolation within several species in the white-headed gull complex. This suggests that just a few genes are driving the speciation process.

Molecular Phylogenetics and Evolution↗

Sediment sources and connectivity linked to hydrologic pathways and geomorphic processes: A conceptual model to specify sediment sources and pathways through space and time

Sediment connectivity is a conceptualization for the transfer and storage of sediment among different geomorphic compartments across upland landscapes and channel networks. Sediment connectivity and dysconnectivity are linked to the water cycle and hydrologic systems with the associated multiscale interactions with climate, soil, topography, ecology, and landuse/landcover under natural variability and human intervention. We review current sediment connectivity and modeling approaches evaluating and quantifying water and sediment transfer in catchment systems. Many studies highlight the interaction between sediment and water in defining landscape connectivity, but many efforts to quantify and/or simulate sediment connectivity rely on the topographic/structural controls on sediment erosion and delivery. More recent modeling efforts integrate functional and structural connectivity to capture hydrologic properties influencing sediment delivery. Though the recent modeling development is encouraging, a comprehensive sediment connectivity framework, which integrates geomorphic and hydrologic processes across spatiotemporal scales, has not yet been accomplished. Such an effort requires understanding the hydrologic and geomorphic processes that control sediment source, storage, and transport at different spatiotemporal scales and across various geophysical conditions. We propose a path for developing this new understanding through an integrated hydrologic and sediment connectivity conceptual model that broadly categorizes dominant processes and patterns relevant to understanding sediment flux dynamics. The conceptual model describes hydrologic–sediment connectivity regimes through spatial-temporal feedback between hydrologic processes and geomorphic drivers. We propose that in combining hydrologic and sediment connectivity into a single conceptual model, patterns emerge such that catchments will exist in a single characteristic behavior at a particular instance, which would shift with space and time, and with landscape disturbances. Using the conceptual model as a “thinking” tool, we extract case studies from a multidisciplinary literature review—from hydrology, geomorphology, biogeochemistry, and watershed modeling to remote-sensing technology—that correspond to each of the dominant hydrologic–sediment connectivity regimes. Sediment and water interactions in real-world examples through various observational and modeling techniques illustrate the advancements in the spatial and temporal scales of landscape connectivity observations and simulations. The conceptual model and case studies provide a foundation for advancing the understanding and predictive capability of watershed sediment processes at multiple spatiotemporal scales. Plain language summary: Soil erosion and movement across the landscape are closely linked to rain events and flow pathways. Landscape connectivity is a way to consider how soil erosion from different parts of the landscape is connected to the streams. We explore where soil erosion occurs and how eroded soil moves across the landscape through the interaction with rainfall and drainage. The comprehensive understanding of sediment connectivity and its dependence on rainfall characteristics and watershed hydrology may help to inform the effective distribution of conservation funds and management actions to address water pollution from excess sediment.

Frontiers in Water↗

Does large dam removal restore downstream riparian vegetation diversity? Testing predictions on the Elwha River, Washington, USA

Large dams and their removal can profoundly affect riparian ecosystems by altering flow and sediment regimes, hydrochory, and landform dynamics, yet few studies have documented these effects on downstream plant communities. Ecological theory and empirical results suggest that by altering disturbance regimes, reducing hydrochory, and shifting communities to later successional stages, dams reduce downstream plant diversity. Dam removal could reverse these processes, but the release of large volumes of sediment could have unexpected, transient effects. Two large dams were removed on the Elwha River in Washington State, USA, from 2011-2014, representing an unprecedented opportunity to study large dam removal effects on riparian plant communities. Our research objectives were to determine: 1) whether the Elwha River dams were associated with lower downstream plant diversity and altered species composition across riparian landforms pre-dam removal, and 2) whether dam removal has begun to restore downstream diversity and composition. To address these objectives, we compared plant species richness and community composition in river segments above, below, and between the two dams. Plant communities were sampled twice before (2005 and 2010) and four times after (2013, 2014, 2016, and 2017) the start of dam removal, with 2013 and 2014 sampled while the upstream dam removal was ongoing. Prior to dam removal, native species richness was 41% lower below dams compared to the upstream segment; six years after dam removal began, it increased ~31% between the dams, while nonnative species richness and cover were not apparently affected by dams or their removal. Deposition caused by large volumes of released reservoir sediment had mixed effects on native species richness (increased on floodplains, decreased elsewhere) in the lowest river segment. Plant community composition was also different downstream from dams compared to the upstream reference, and has changed in downstream floodplains and bars since dam removal. Long-term, we expect that diversity will continue to increase in downstream river segments. Our results provide evidence that 1) large dams reduce downstream native plant diversity, 2) dam removal may restore it, and 3) given the natural dynamics of riparian vegetation, long-term, multi-year before-and-after-monitoring is essential for understanding dam removal effects.

Washington↗

Hydrological connectivity for riverine fish: measurement challenges and research opportunities

In this review, we first summarize how hydrologic connectivity has been studied for riverine fish capable of moving long distances, and then identify research opportunities that have clear conservation significance. Migratory species, such as anadromous salmonids, are good model organisms for understanding ecological connectivity in rivers because the spatial scale over which movements occur among freshwater habitats is large enough to be easily observed with available techniques; they are often economically or culturally valuable with habitats that can be easily fragmented by human activities; and they integrate landscape conditions from multiple surrounding catchment(s) with in‐river conditions. Studies have focussed on three themes: (i) relatively stable connections (connections controlled by processes that act over broad spatio‐temporal scales >1000 km 2 and >100 years); (ii) dynamic connections (connections controlled by processes acting over fine to moderate spatio‐temporal scales ∼1–1000 km 2 and <1–100 years); and (iii) anthropogenic influences on hydrologic connectivity, including actions that disrupt or enhance natural connections experienced by fish. We outline eight challenges to understanding the role of connectivity in riverine fish ecology, organized under three foci: (i) addressing the constraints of river structure; (ii) embracing temporal complexity in hydrologic connectivity; and (iii) managing connectivity for riverine fishes. Challenges include the spatial structure of stream networks, the force and direction of flow, scale‐dependence of connectivity, shifting boundaries, complexity of behaviour and life histories and quantifying anthropogenic influence on connectivity and aligning management goals. As we discuss each challenge, we summarize relevant approaches in the literature and provide additional suggestions for improving research and management of connectivity for riverine fishes. Specifically, we suggest that rapid advances are possible in the following arenas: (i) incorporating network structure and river discharge into analyses; (ii) increasing explicit consideration of temporal complexity and fish behaviour in the scope of analyses; and (iii) parsing degrees of human and natural influences on connectivity and defining acceptable alterations. Multiscale analyses are most likely to identify dominant patterns of connections and disconnections, and the appropriate scale at which to focus conservation activities.

Freshwater Biology↗

Ecological factors decouple Great Lakes fish mercury concentrations trends decadal declines in mercury emissions

Atmospheric mercury (Hg) deposition has been declining in North America but remains the dominant delivery mechanism to the Great Lakes. The Lakes are highly efficient at bioaccumulating methylmercury, making the fish excellent sentinels for tracking shifts in atmospheric Hg deposition. Invasive mussels have altered biogeochemical processes, prey populations and fish dietary strategies asynchronously and to varied extents across the lower four lakes, impacting fish Hg exposure. To test if fish are adapting to new biogeochemical conditions, we analyzed a 40 year fish archive for carbon and nitrogen isotope ratios and amino acid-specific nitrogen isotope ratios. To assess Hg sources, we measured Hg isotope ratios. We reconstructed and compared energetic pathways that impact fish Hg concentrations to Hg-source trends. We found fish-Hg concentrations are declining but not monotonically due to ecological disturbances. Fish-Hg isotope values, unimpacted by ecological disturbance, confirm that sources of bioaccumulated Hg shift contemporaneously with changes in atmospheric Hg concentrations. Across Lakes, the degree of responsiveness to changes in atmospheric Hg concentrations mirrors the proportion of atmospheric-delivered Hg we previously modeled. Changes in both fish concentrations and fish isotope values outpace paleolimnetic reconstructions suggesting declines in atmospheric Hg concentrations impact fish Hg more than sediment.

Great Lakes↗

Concordant patterns of morphological, stable isotope, and genetic variation in a recent ecological radiation (Salmonidae: Coregonus spp.)

Groups of sympatric taxa with low interspecific genetic differentiation, but considerable ecological differences, offer great opportunities to study the dynamics of divergence and speciation. This is the case of ciscoes ( Coregonus spp.) in the Laurentian Great Lakes, which are characterized by a complex evolutionary history and are commonly described as having undergone an adaptive radiation. In this study, morphometrics, stable isotopes and transcriptome sequencing were used to study the relationships within the Coregonus artedi complex in western Lake Superior. We observed general concordance for morphological, ecological and genomic variation, but the last was more taxonomically informative as it showed less overlap among species in multivariate space. Low levels of genetic differentiation were observed between individuals morphologically identified as Coregonus hoyi and C. zenithicus , which could be evidence of incomplete lineage sorting or recent hybridization between the two groups. Transcriptome-based single nucleotide polymorphisms exhibited significant divergence for genes associated with vision, development, metabolism and immunity among species that occupy different habitats. This study highlights the importance of using an integrative approach when studying groups of taxa with a complex evolutionary history, as individual-level analyses of multiple independent data sets can provide a clearer picture of the patterns and processes associated with the origins of biodiversity.

Michigan, Minnesota, Ontario, Wisconsin↗

Using a structured decision analysis to evaluate bald eagle vital signs monitoring in Southwest Alaska National Parks

1. Monitoring programs can benefit from an adaptive monitoring approach, where key decisions about why, where, what, and how to monitor are revisited periodically in order to ensure programmatic relevancy. 2. The National Park Service (NPS) monitors status and trends of Vital Signs to evaluate compliance with the NPS mission. Although abundant, The Southwest Alaska Network (SWAN) monitors bald eagles because of their inherent importance to park visitors and role as an important ecological indicator. Our goal is to identify an optimal monitoring program that may be standardized among participating parks. 3. We gathered an expert panel of scientists and managers, and implemented a Delphi Process to gather information about the bald eagle monitoring program. Panelists generated a list of means objectives for the monitoring program: minimizing cost, minimizing effort, maximizing the ability to detect change in bald eagle populations, and maximizing the amount of accurate information collected about bald eagles. 4. We used a swing-weighting technique to assign importance to each objective. Collecting accurate information about bald eagles was considered the most important means objective. 5. Combining panelist-generated information with objective importance, we analyzed the scenarios and defined the optimal decision using linear value modeling. Through our analysis, we found that a “Comprehensive” monitoring scenario, comprised of all feasible monitoring metrics is the optimal monitoring scenario. Even with greatly increased cost, the Comprehensive monitoring scenario remains the best solution. 6. We suggest further exploration of the cost and effort required for the Comprehensive scenario, to determine if it is in the parks’ best interest to begin monitoring additional metrics.

Alaska↗

Performance evaluation of a channel rehabilitation project on the Lower Missouri River and implications for the dispersal of larval pallid sturgeon

In the Lower Missouri River, extensive channel modifications have altered hydraulic and morphologic conditions and reduced the river's ecological integrity. One species that has been adversely affected by these changes is the pallid sturgeon ( Scaphirhynchus albus ). Mainstem dams on the Missouri River restrict the upstream migration of adults and limit the downstream dispersal of larvae. Channelization to facilitate commercial barge traffic has also simplified the river. The self-dredging navigation channel is a highly efficient conduit for transporting sand, which has resulted in diminished rearing habitat along the lower river. Recently, a series of experimental projects was implemented to reengineer selected bends of the Lower Missouri River with the goal of increasing the interception and retention of passively drifting age-0 sturgeon into habitats more conducive to rearing. Here, we evaluate the hydraulic performance of one of these rehabilitation projects to gain insight on the implications of these interventions for age-0 pallid sturgeon dispersal. We conducted a dye-trace experiment and complementary hydraulic and particle-tracking modeling to examine the spatial and temporal patterns of passive dispersal in and around the rehabilitated study reach. Results from both the dye-trace experiment and particle-tracking model highlight the presence of several interception pathways from the navigation channel into more suitable rearing habitat on channel margins. Moreover, our results indicate that residence times within the rearing habitat are increased in comparison to the main channel. Although we cannot provide biological evaluation at this time to assess whether the rehabilitated study bend intercepts passively drifting age-0 pallid sturgeon, our analysis shows that hydraulic conditions within the rehabilitated bend would favor interception and retention of passively drifting particles (or, presumably, larvae) from the navigation channel and into slower moving, shallow-water habitat. Moreover, our particle-tracking model provides a new capability to explore important biological transport processes across a range of flows, organisms, and river environments.

Lower Missouri River↗

Time to get real with qPCR controls: The frequency of sample contamination and the informative power of negative controls in environmental DNA studies

Environmental (e)DNA methods have enabled rapid, sensitive and specific inferences of taxa presence throughout diverse fields of ecological study. However, use of eDNA results for decision-making has been impeded by uncertainties associated with false positive tests putatively caused by sporadic or systemic contamination. Sporadic contamination is a process that is inconsistent across samples and systemic contamination occurs consistently over a group of samples. Here, we used empirical data and laboratory experiments to (i) estimate the sporadic contamination rate for each stage of a common, targeted eDNA workflow employing best practice quality control measures under simulated conditions of rare and common target DNA presence, (ii) determine the rate at which negative controls (i.e., “blanks”) detect varying concentrations of systemic contamination, and (iii) estimate the effort that would be required to consistently detect sporadic and systemic contamination. Sporadic contamination rates were very low across all eDNA workflow steps, and, therefore, an intractably high number of negative controls (>100) would be required to determine occurrence of sporadic contamination with any certainty. Contrarily, detection of intentionally introduced systemic contamination was more consistent; therefore, very few negative controls (<5) would be needed to consistently alert to systemic contamination. These results have considerable implications to eDNA study design when resources for sample analyses are constrained.

Molecular Ecology Resources↗