Environmental DNA (eDNA) surveys have become important tools for monitoring aquatic biodiversity. Barcode sequencing of eDNA generates community profiles that, while potentially biased in both capture and amplification, can nonetheless yield high information content per unit cost. While factors affecting eDNA capture and amplification have been heavily studied, watershed-scale assessments of fish communities and our confidence in such have been less frequent. We performed an initial watershed-scale characterization of fish eDNA using rapid, low-volume filtering with replicate and control samples scaled for a single Illumina MiSeq flow cell, using the mitochondrial 12S ribosomal RNA locus for taxonomic profiling. Our bioinformatic approach included 1) direct estimation of sequencing error from unambiguous mappings (alignments) and simulation of error in taxonomic assignment under various mapping criteria; 2) binning of species based on inferred assignment error rather than by taxonomic rank; and 3) visualization of mismatch distributions to facilitate discovery of distinct haplotypes attributed to the same reference. Our approach was implemented for the St. Regis River, New York, United States, which supports a valuable recreational fishery and has been a target of restoration activities. We used a large record of St. Regis-specific observations to validate our assignments. We found that 300 mL drawn through 25-mm filters yielded greater than 5 ng/µL DNA at most sites in August and September, which was an approximate threshold for generating strong sequencing libraries in our hands. Using inferred sequence error rates, we binned 12S references for 110 species on a state-level checklist into 85 single-species bins and seven multispecies bins. Of 48 taxonomic bins actually observed in the St. Regis, we detected eDNA consistent with 40, with an additional four detections flagged as potential contaminants post-collection. Sixteen unobserved species detected by eDNA ranged from plausible to implausible based on distributional data, whereas six observed species had no 12S reference sequence.

Environmental DNA (eDNA) surveys have become important tools for monitoring aquatic biodiversity. Barcode sequencing of eDNA generates community profiles that, while potentially biased in both capture and amplification, can nonetheless yield high information content per unit cost. While factors affecting eDNA capture and amplification have been heavily studied, watershed-scale assessments of fish communities and our confidence in such have been less frequent. We performed an initial watershed-scale characterization of fish eDNA using rapid, low-volume filtering with replicate and control samples scaled for a single Illumina MiSeq flow cell, using the mitochondrial 12S ribosomal RNA locus for taxonomic profiling. Our bioinformatic approach included 1) direct estimation of sequencing error from unambiguous mappings (alignments) and simulation of error in taxonomic assignment under various mapping criteria; 2) binning of species based on inferred assignment error rather than by taxonomic rank; and 3) visualization of mismatch distributions to facilitate discovery of distinct haplotypes attributed to the same reference. Our approach was implemented for the St. Regis River, New York, United States, which supports a valuable recreational fishery and has been a target of restoration activities. We used a large record of St. Regis-specific observations to validate our assignments. We found that 300 mL drawn through 25-mm filters yielded greater than 5 ng/µL DNA at most sites in August and September, which was an approximate threshold for generating strong sequencing libraries in our hands. Using inferred sequence error rates, we binned 12S references for 110 species on a state-level checklist into 85 single-species bins and seven multispecies bins. Of 48 taxonomic bins actually observed in the St. Regis, we detected eDNA consistent with 40, with an additional four detections flagged as potential contaminants post-collection. Sixteen unobserved species detected by eDNA ranged from plausible to implausible based on distributional data, whereas six observed species had no 12S reference sequence.

In 1991, the U.S. Geological Survey (USGS) began a study of more than 50 major river basins across the Nation as part of the National Water-Quality Assessment (NAWQA) project of the National Water Quality Program. One of the major goals of the NAWQA project is to determine how water quality and ecological conditions change over time. To support that goal, long-term consistent and comparable ecological monitoring has been conducted on streams and rivers throughout the Nation. Fish, invertebrate, and algae data collected as part of the NAWQA program were retrieved from the USGS Aquatic Bioassessment database for use in trend analysis. Ultimately, these data will provide insight into how natural features and human activities have contributed to changes in ecological condition over time in the Nation’s streams and rivers. This USGS data release contains all of the input and output files necessary to reproduce the results of the ecological trend analysis described in the associated U.S. Geological Survey Scientific Investigations Report. Data preparation for input to the model is also fully described in the above mentioned report.

In 1991, the U.S. Geological Survey (USGS) began a study of more than 50 major river basins across the Nation as part of the National Water-Quality Assessment (NAWQA) project of the National Water Quality Program. One of the major goals of the NAWQA project is to determine how water quality and ecological conditions change over time. To support that goal, long-term consistent and comparable ecological monitoring has been conducted on streams and rivers throughout the Nation. Fish, invertebrate, and algae data collected as part of the NAWQA program were retrieved from the USGS Aquatic Bioassessment database for use in trend analysis. Ultimately, these data will provide insight into how natural features and human activities have contributed to changes in ecological condition over time in the Nation’s streams and rivers. This USGS data release contains all of the input and output files necessary to reproduce the results of the ecological trend analysis described in the associated U.S. Geological Survey Scientific Investigations Report. Data preparation for input to the model is also fully described in the above mentioned report.

A survey of environmental DNA was performed in Tunison Creek downstream of the USGS Tunison Aquatic Laboratory. The goal of the survey was to characterize the source ecological community at multiple trophic or taxonomic levels by associating DNA fragments with reference databases. Three taxonomically informative genetic loci were used: the mitochondrial cytochrome oxidase 1 locus, the mitochondrial 12S ribosomal locus, and the bacterial/organellar 16S ribosomal locus. The data set includes a list of sample and sequence accessions residing in the National Center for Biotechnology Information databases, which provide the raw data for analysis or re-use. The data also includes the representative sequences of operational taxonomic units (OTUs) and their abundance and taxonomic assignment, which are the primary endpoints of the data collection.

A survey of environmental DNA was performed in Tunison Creek downstream of the USGS Tunison Aquatic Laboratory. The goal of the survey was to characterize the source ecological community at multiple trophic or taxonomic levels by associating DNA fragments with reference databases. Three taxonomically informative genetic loci were used: the mitochondrial cytochrome oxidase 1 locus, the mitochondrial 12S ribosomal locus, and the bacterial/organellar 16S ribosomal locus. The data set includes a list of sample and sequence accessions residing in the National Center for Biotechnology Information databases, which provide the raw data for analysis or re-use. The data also includes the representative sequences of operational taxonomic units (OTUs) and their abundance and taxonomic assignment, which are the primary endpoints of the data collection.

In 1991, the U.S. Geological Survey (USGS) began a study of more than 50 major river basins across the Nation as part of the National Water-Quality Assessment (NAWQA) project of the National Water Quality Program. One of the major goals of the NAWQA project is to determine how water quality and ecological conditions change over time. To support that goal, long-term consistent and comparable ecological monitoring has been conducted on streams and rivers throughout the Nation. Fish, invertebrate, and algae data collected as part of the NAWQA program were retrieved from the USGS Aquatic Bioassessment database for use in trend analysis. Ultimately, these data will provide insight into how natural features and human activities have contributed to changes in ecological condition over time in the Nation’s streams and rivers. This dataset is a model archive containing all input files and R source code to reproduce the results of the ecological trend analysis described in the associated U.S. Geological Survey Scientific Investigations Report. Associated output files are also included in the archive. Data preparation for input to the model is fully described in the above mentioned report.

In 1991, the U.S. Geological Survey (USGS) began a study of more than 50 major river basins across the Nation as part of the National Water-Quality Assessment (NAWQA) project of the National Water Quality Program. One of the major goals of the NAWQA project is to determine how water quality and ecological conditions change over time. To support that goal, long-term consistent and comparable ecological monitoring has been conducted on streams and rivers throughout the Nation. Fish, invertebrate, and algae data collected as part of the NAWQA program were retrieved from the USGS Aquatic Bioassessment database for use in trend analysis. Ultimately, these data will provide insight into how natural features and human activities have contributed to changes in ecological condition over time in the Nation’s streams and rivers. This dataset is a model archive containing all input files and R source code to reproduce the results of the ecological trend analysis described in the associated U.S. Geological Survey Scientific Investigations Report. Associated output files are also included in the archive. Data preparation for input to the model is fully described in the above mentioned report.

The data released are associated with an examination of eDNA from round goby fish (Neogobius melanostomus) in a series of field sample collections (lake nearshore and stream transport) and in vitro laboratory experiments. The round goby was used as a model for our source of eDNA in the field collections and mesocosm experiments. The field samples for lake nearshore (water and sediment) were collected from two Lake Michigan shoreline locations (Portage Lakefront, Portage, Indiana and Washington Park, Michigan City, Indiana) to examine the detectability of eDNA, the influence of sediment on eDNA estimates, and eDNA spatial and temporal resolution. The field samples for stream transport (water) were collected from Brown Ditch, a Lake Michigan stream, and were used to model and test eDNA transport. Rates for eDNA shedding and decay in water and sediment were developed using in vitro laboratory experiments and provided a comparison of eDNA behavior (persistence and degradation) and detection in water and sediment.