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Patrick R. Hutchins

Publications and source records attributed to Patrick R. Hutchins.

12 recordsLinked to original sources

Temporal replication and sampling efficiency in high-frequency autonomous and manual eDNA sampling

Environmental DNA (eDNA) sampling is a valuable tool for ecological research, but its reliability as a quantitative approach can be improved by accounting for unexplained spatiotemporal variability and autocorrelation in eDNA concentrations. Sampling designs with temporal replication can separate sampling variation from temporal variation, but obtaining true temporal replicates is difficult when samples are collected sequentially. Autosamplers may facilitate high-frequency eDNA sampling, but their sequential sampling structure raises questions about when closely spaced samples can reasonably be treated as temporal replicates. This 19.5-day study evaluated temporal variation in invasive Round Goby ( Neogobius melanostomus ) eDNA concentration at a single large-river site with short-term hydrologic variability. We used two synchronized autosamplers and a backpack sampler to collect samples as frequently as every 15 min using polyethersulfone (PES) or mixed cellulose ester (MCE) filters. We fit hierarchical temporal models that decomposed sampling and temporal variation, evaluated evidence for residual temporal correlation under alternative priors for the effective range, and used simulation to assess bias when closely spaced sequential samples were treated as simultaneous temporal replicates. We found evidence of residual temporal correlation at short lag scales, but the effective range of temporal correlation was weakly identified and sensitive to prior assumptions. Hydrologic covariates had limited explanatory support once temporal structure was accounted for. Simulations showed that treating sequential samples collected over 15–60 min windows as simultaneous temporal replicates can bias estimates of sampling and temporal variation, especially when temporal correlation is relatively weak. MCE filters produced higher expected recovered copy numbers than PES filters, while evidence for systematic differences among sampler types was weaker. These results emphasize that eDNA temporal replication should be collected as close together in time as possible, and that uncertainty in temporal correlation should be explicitly considered when designing and analyzing high-frequency eDNA surveys.

New York↗

Accounting for temporal variation and correlation in environmental DNA sampling can improve ecological inferences

Environmental DNA (eDNA) concentration varies through space and time, and measurements collected close together are often correlated. Ignoring this dependence can inflate the rate of incorrect ecological inferences (Type I error rate). Although spatial correlation in eDNA has received considerable attention, temporal correlation has been less well studied. Statistical models and study designs that account for temporal correlation are increasingly important to understand time-dependent effects in complex systems. We developed a hierarchical model that separates temporal ecological variation from variability stemming from sampling and laboratory processes and applied it to four single-site eDNA time series collected over 17–24 days, three of which provided sufficient information for parameter estimation. We then used the empirically estimated parameter magnitudes in a simulation study to evaluate alternative temporal sampling designs that considered (1) how a fixed number of samples are allocated across different numbers of sampling times with different levels of temporal replication and (2) equally spaced versus cluster-spaced sampling (short bursts separated by longer gaps). Across the three time series sufficient for analysis, we observed substantial sampling variability, temporal variability, and temporal correlation, although correlation was estimated imprecisely (large coefficient of variation). Simulations showed that when sampling intervals were shorter than the effective temporal correlation range, models that ignored temporal dependence produced inflated Type I error rates and frequently detected spurious temporal trends. Accounting for temporal correlation substantially reduced this inflated Type I error rate. Optimal sampling strategies depended on study objectives. Clustered sampling most effectively estimated temporal correlation. When temporal dependence was negligible, evenly spaced sampling maximized power to detect trends. Estimating sampling variability required concentrating effort into fewer sampling times with more replicates per time, whereas estimating temporal variance was most precise with intermediate levels of replication. Together, these results indicate that temporal dependence can strongly affect inference from quantitative eDNA time series when sampling intervals approach the correlation timescale. Designs that ignore this dependence risk inferring ecological change or difference where none exists. Our framework provides practical guidance for allocating sampling effort in temporally intensive eDNA monitoring and for interpreting trends from short time series.

Idaho, Florida, Montana, Wisconsin↗

Bright spot in eDNA monitoring: Early detection of invasive New Zealand mudsnails (Potamopyrgus antipodarum) prompted effective rapid response for fish hatchery

The New Zealand mudsnail (NZMS; Potamopyrgus antipodarum ) is a widespread aquatic invasive species that is parthenogenic, requiring only a single individual to initiate an infestation. Fish hatcheries–which are critical infrastructure that raise fish to support conservation, recreation, and subsistence fisheries–frequently use local water sources to provide cool water and are especially vulnerable to NZMS invasion from the contamination of water supplies. If an invasion proceeds undetected, hatcheries pose a risk for compounding the spread of NZMS because their operations transfer live organisms and associated water between hatchery facilities and, when stocking, to rivers and lakes. The U.S. Fish and Wildlife Service's Alchesay National Fish Hatchery, located on the Fort Apache Indian Reservation in Whiteriver, Arizona, produces trout to stock in Tribal reservoirs, lakes, and rivers across the southwestern U.S. New Zealand mudsnails were first documented in Arizona in 1995, are now widespread in this region, and occur at the confluence of the hatchery's outflow with the North Fork White River. Contamination of water supplies is the principal pathway for NZMS invasion into the hatchery. Here, we describe early detection environmental DNA (eDNA) surveillance efforts for NZMS at Alchesay National Fish Hatchery. Positive eDNA detections initiated a chain of events that ultimately led to four NZMS individuals being discovered and a rapid response eradication effort. Follow-up eDNA sampling and visual observation efforts after the eradication effort have yielded no detections of NZMS eDNA. We credit the success of this case with four key elements: rapid turnaround times, a robust quality assurance scheme, a proactive eDNA sampling design, and established partnerships. To our knowledge, this is the first published case of eDNA monitoring being used for early detection and successful rapid response for complete removal of an invasive species in a fish hatchery.

Arizona↗

Effects of supplemental feeding on the fecal bacterial communities of Rocky Mountain elk in the Greater Yellowstone Ecosystem

Supplemental feeding of wildlife is a common practice often undertaken for recreational or management purposes, but it may have unintended consequences for animal health. Understanding cryptic effects of diet supplementation on the gut microbiomes of wild mammals is important to inform conservation and management strategies. Multiple laboratory studies have demonstrated the importance of the gut microbiome for extracting and synthesizing nutrients, modulating host immunity, and many other vital host functions, but these relationships can be disrupted by dietary perturbation. The well-described interplay between diet, the microbiome, and host health in laboratory and human systems highlights the need to understand the consequences of supplemental feeding on the microbiomes of free-ranging animal populations. This study describes changes to the gut microbiomes of wild elk under different supplemental feeding regimes. We demonstrated significant cross-sectional variation between elk at different feeding locations and identified several relatively low-abundance bacterial genera that differed between fed versus unfed groups. In addition, we followed four of these populations through mid-season changes in supplemental feeding regimes and demonstrated a significant shift in microbiome composition in a single population that changed from natural forage to supplementation with alfalfa pellets. Some of the taxonomic shifts in this population mirrored changes associated with ruminal acidosis in domestic livestock. We discerned no significant changes in the population that shifted from natural forage to hay supplementation, or in the populations that changed from one type of hay to another. Our results suggest that supplementation with alfalfa pellets alters the native gut microbiome of elk, with potential implications for population health.

Wyoming↗

Integrating environmental DNA results with diverse data sets to improve biosurveillance of river health

Autonomous, robotic environmental (e)DNA samplers now make it possible for biological observations to match the scale and quality of abiotic measurements collected by automated sensor networks. Merging these automated data streams may allow for improved insight into biotic responses to environmental change and stressors. Here, we merged eDNA data collected by robotic samplers installed at three U.S. Geological Survey (USGS) streamgages with gridded daily weather data, and daily water quality and quantity data into a cloud-hosted database. The eDNA targets were a rare fish parasite and a more common salmonid fish. We then used computationally expedient Bayesian hierarchical occupancy models to evaluate associations between abiotic conditions and eDNA detections and to simulate how uncertainty in result interpretation changes with the frequency of autonomous robotic eDNA sample collection. We developed scripts to automate data merging, cleaning and analysis steps into a chained-step, workflow. We found that inclusion of abiotic covariates only provided improved insight for the more common salmonid fish since its DNA was more frequently detected. Rare fish parasite DNA was infrequently detected, which caused occupancy parameter estimates and covariate associations to have high uncertainty. Our simulations found that collecting samples at least once per day resulted in more detections and less parameter uncertainty than less frequent sampling. Our occupancy and simulation results together demonstrate the advantages of robotic eDNA samplers and how these samples can be combined with easy to acquire, publicly available data to foster real-time biosurveillance and forecasting.

Idaho, Wyoming, Montana↗

The elephant in the lab (and field): Contamination in aquatic environmental DNA studies

The rapid evolution of environmental (e)DNA methods has resulted in knowledge gaps in smaller, yet critical details like proper use of negative controls to detect contamination. Detecting contamination is vital for confident use of eDNA results in decision-making. We conducted two literature reviews to summarize (a) the types of quality assurance measures taken to detect contamination of eDNA samples from aquatic environments, (b) the occurrence, frequency and attribution (i.e., putative sources) of unexpected amplification in these quality assurance samples, and (c) how results were interpreted when contamination occurred. In the first literature review, we reviewed 156 papers and found that 91% of targeted and 73% of metabarcoding eDNA studies reported inclusion of negative controls within their workflows. However, a large percentage of targeted (49%) and metabarcoding (80%) studies only reported negative controls for laboratory procedures, so results were potentially blind to field contamination. Many of the 156 studies did not provide critical methodological information and amplification results of negative controls. In our second literature review, we reviewed 695 papers and found that 30 targeted and 32 metabarcoding eDNA studies reported amplification of negative controls. This amplification occurred at similar proportions for field and lab workflow steps in targeted and metabarcoding studies. These studies most frequently used amplified negative controls to delimit a detection threshold above which is considered significant or provided rationale for why the unexpected amplifications did not affect results. In summary, we found that there has been minimal convergence over time on negative control implementation, methods, and interpretation, which suggests that increased rigor in these smaller, yet critical details remains an outstanding need. We conclude our review by highlighting several studies that have developed especially effective quality assurance, control and mitigation methods.

Frontiers in Ecology and Evolution↗

A round-robin evaluation of the repeatability and reproducibility of environmental DNA assays for dreissenid mussels

Resource managers may be hesitant to make decisions based on environmental (e)DNA results alone since eDNA is an indirect method of species detection. One way to reduce the uncertainty of eDNA is to identify laboratory‐based protocols that ensure repeatable and reproducible results. We conducted a double‐blind round‐robin analysis of probe‐based assays for DNA of dreissenid ( Dreissena spp.) mussels, which are prolific aquatic invaders that can cause significant economic and ecological impacts. DNA extract from water samples spiked with known amounts of dreissenid DNA and from water samples collected from waters with and without dreissenids were analyzed by four independent research laboratories. We used results to calculate detection repeatability within laboratories and assays, detection reproducibility among laboratories and assays, and estimated dreissenid DNA copy number precision and accuracy. Laboratory and assay repeatability and reproducibility of detection results were high, 91% and 92%, respectively. The estimated copy numbers were neither precise nor accurate for samples spiked with <773 gene copies. These results suggest that eDNA surveillance of dreissenid mussels, using the protocols evaluated herein, can generate reliable detection data for decision‐making. However, managers should be cautious about using the quantitative information often associated with eDNA detections, especially when DNA is at lower abundance. Our results provide strong support that eDNA has the potential to provide repeatable and reproducible evidence under varying laboratory conditions and for different sample water chemistries. This is reassuring since the demand for eDNA surveillance is widespread and number of laboratories that process eDNA samples is growing steadily.

California, Montana, Nevada, New York, Oregon, Was↗

Adding invasive species bio-surveillance to the U.S. Geological Survey streamgage network

The costs of invasive species in the United States alone are estimated to exceed US$100 billion per year so a critical tactic in minimizing the costs of invasive species is the development of effective, early-detection systems. To this end, we evaluated the efficacy of adding environmental (e)DNA surveillance to the U.S. Geological Survey (USGS) streamgage network, which consists of > 8,200 streamgages nationwide systemically visited by USGS hydrologic technicians. Incorporating strategic eDNA sample collection during routine streamgage visits could provide early detection surveillance of aquatic invasive species with minimal additional cost. For this evaluation, USGS hydrologic technicians collected monthly eDNA water samples, May – September 2018, from streamgages downstream of reservoirs in the Columbia River Basin thought to be vulnerable to invasive dreissenid mussel ( Dreissenidae spp.) establishment. We tested water samples for dreissenid mussel DNA and also for kokanee ( Oncorhynchus nerka ) and yellow perch ( Perca flavescens ) DNA; the two fishes were used to assess if streamgages are adequately located to provide early-detection eDNA surveillance of taxa known to be present in upstream reservoirs. No Columbia River Basin streamgage samples met our criteria for being scored as positive for dreissenid DNA. We did detect kokanee and yellow perch DNA at all streamgages downstream of reservoirs where these species are known to occur. Field collection, laboratory analyses, and personnel time required for collection of four eDNA samples at a streamgage site cost US$500 -US$600 (net). Given these results, incorporating eDNA biosurveillance into routine streamgage visits might decrease costs associated with an invasion since early detection maximizes the potential for eradication, containment, and mitigation.

Ecosphere↗

The Yellowstone River fish-kill: Fish health informs and is informed by vital signs monitoring

Trout are socioeconomically and ecologically important in the Greater Yellowstone Area (GYA); yet these fish face numerous threats. Disease may begin to play a larger role in reducing fish populations, partly because many existing threats may interact to exacerbate the frequency, extent, and severity of fish diseases (Lafferty 2009). For example, habitat loss and low summer flows might interact to stress fish, making them more susceptible to disease while also increasing fish densities in microhabitats, thereby creating conditions where infectious diseases are more easily spread. Conservation and management efforts to mitigate these threats often involve reactionary measures to unforeseen events. Long-term monitoring of aquatic vital signs and fish health, however, may provide important insights for predicting the spread of fish diseases and the extent and severity of outbreaks.

Idaho, Montana, Wyoming↗

Improved conventional PCR assay for detecting Tetracapsuloides bryosalmonae DNA in fish tissues

Conventional PCR is an established method to detect Tetracapsuloides bryosalmonae DNA in fish tissues and to confirm diagnosis of proliferative kidney disease (PKD) caused by T. bryosalmonae . However, the commonly used PKX5f‐6r primers were designed with the intention of obtaining sequence information and are suboptimal for determining parasite DNA presence. A new PCR assay to detect T. bryosalmonae 18s rDNA, PKX18s1266f‐1426r, is presented that demonstrates specificity, repeatability, and enhanced sensitivity over the PKX5f‐6r assay. The limit of detection of the PKX18s1266f‐1426r assay at 95% confidence was 100 template copies, and the new primers detected parasite DNA more consistently at template concentrations below 100 copies than did PKX5f‐6r. The PKX18s1266f‐1426r also achieved 100% detection at sample DNA concentrations one order of magnitude lower than PKX5f‐6r. Out of 127 salmonid fish with unknown T. bryosalmonae infection status, PKX5f‐6r detected 35 positive samples, while the new assay detected 43. The discrepancy in T. bryosalmonae detection between the two primer sets may be attributed to several differences between the assays, including oligonucleotide melting temperatures, the use of a touchdown PCR thermal cycle, and amplicon length.

Journal of Aquatic Animal Health↗

Tradeoffs of a portable, field-based environmental DNA platform for detecting invasive northern pike (Esox lucius) in Alaska

Environmental DNA (eDNA) has improved detection probabilities of aquatic invasive species but lab-based analytical platforms for eDNA analyses slow opportunities for rapid response. Effective approaches that address this analytical bottleneck and improve capacity for rapid response are urgently needed. We tested the sensitivity of a portable, field-based eDNA platform relative to widely used lab-based eDNA approaches for detecting invasive northern pike (Esox lucius) in eight lakes on Alaska’s Kenai Peninsula. The portable, field-based platform takes ~ 1 hr from sample collection to final results and uses a field-based DNA extraction kit, a shelf-stable assay, and a portable real-time PCR thermocycler. Lab-based approaches take days to weeks to months for final results and use lab-based DNA extraction kits, lab-bound assays, and benchtop real-time thermocyclers. We found that the portable, field-based approach was less sensitive than lab-based approaches and was more prone to inhibition, thus increasing potential for false-negatives. Until sensitivity and inhibition issues can be resolved, this portable, field-based approach is best viewed as a complement to rather than a replacement of standard eDNA lab-based approaches.

Alaska↗

A probe-based quantitative PCR assay for detecting Tetracapsuloides bryosalmonae in fish tissue and environmental DNA water samples

A probe-based quantitative real-time PCR assay was developed to detect Tetracapsuloides bryosalmonae , which causes proliferative kidney disease in salmonid fish, in kidney tissue and environmental DNA (eDNA) water samples. The limits of detection and quantification were 7 and 100 DNA copies for calibration standards and T. bryosalmonae was reliably detected down to 100 copies in tissue and eDNA samples. The assay presented here is a highly sensitive and quantitative tool for detecting T. bryosalmonae with potential applications for tissue diagnostics and environmental detection.

Conservation Genetics Resources↗