Validation and application of a standardized quantitative PCR assay for the assessment of antimicrobial resistance genes in surface water

Laura Celeste Scott; Christina A. Ahlstrom; Hanna Woksepp; Jonas Bonnedahl; Cherie Marie Mckeeman; Mark E. Miller; Dave Shirokauer; Andrew M. Ramey

doi:10.1038/s41598-026-35635-x

USGS · 70275250

Validation and application of a standardized quantitative PCR assay for the assessment of antimicrobial resistance genes in surface water

Laura Celeste Scott·Christina A. Ahlstrom·Hanna Woksepp·Jonas Bonnedahl·Cherie Marie Mckeeman·Mark E. Miller·Dave Shirokauer·Andrew M. Ramey

View original source ↗Export metadata ↓

Abstract

Antimicrobial resistance can be an indicator of anthropogenic contamination in surface waters and is a potential public health threat. Methodological standardization for characterization of antimicrobial resistance in the environment is lacking. Quantitative PCR (qPCR) is used for rapid assessment of antibiotic resistance genes (ARGs) from environmental sources, including surface water. Here we describe the validation and application of a qPCR assay for 47 bacterial gene targets intended for surface water samples. The qPCR assay displayed excellent sensitivity (97.66%) and specificity (98.71%) for detecting ARGs when compared to whole genome sequencing of bacterial isolates. The qPCR assay was able to detect up to 6/8 (75.0%) of ARGs spiked into sterile water at varying concentrations and four sample ultrafiltration volumes. Nineteen different ARGs were detected across six samples sites at three national parks in Alaska using ultrafiltered surface water samples. The number of unique ARGs detected was higher at sites within parks with greater visitation. The relative abundance of ARGs/16S from Exit Creek in Kenai Fjords National Park, downstream from a visitor center was greater than all other sampled sites. We have demonstrated a robust qPCR assay for monitoring ARGs in surface waters, including those that are minimally human impacted.

Explore related subjects

Scientific Reports Article

Source-reported bounding extent: 58.766526235513254° to 64.323291045561° latitude; -155.99719849243888° to -139.26423702947662° longitude. This indicates report coverage, not an exact sampling location. View area on OpenStreetMap.

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

Laura Celeste Scott, Christina A. Ahlstrom, Hanna Woksepp, Jonas Bonnedahl, Cherie Marie Mckeeman, Mark E. Miller, Dave Shirokauer, Andrew M. Ramey. 2026-03-02. Validation and application of a standardized quantitative PCR assay for the assessment of antimicrobial resistance genes in surface water. https://doi.org/10.1038/s41598-026-35635-x

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related USGS reports

USGS2026

Rewilding the upper Klamath River Basin: Rapid recolonization of Chinook salmon following the world’s largest dam removal

Habitat fragmentation is a global driver of decline in biodiversity, yet evaluations of ecological response following re-connection remain rare. In 2024, four hydroelectric dams on the Klamath River were removed, reconnecting over 640 km of anadromous habitat and marking the largest dam removal project completed to date. We used a stationary high-resolution multi-beam imaging SONAR to quantify adult Chinook salmon ( Oncorhynchus tshawytscha ) passage into newly reopened reaches during the first two migratory seasons following dam removal. An estimated 7,742 (95% CI: 7,702–7,778) Chinook salmon migrated upstream in 2024 and 13,310 (95% CI: 12,876–13,733) in 2025, representing 18–19% of fish returning to the Klamath River Basin. Within this period, Chinook salmon recolonized 88% of their documented historical distribution, expanding into 345 km of reconnected habitats and reaching elevations of up to 1,250 m. Recolonization required fish to swim through dam removal sites, complex hydraulic corridors and engineered fishways, demonstrating the propensity for rapid recolonization at basin scale despite these challenges. These findings provide the first quantitative assessment of salmon response to the world’s largest dam removal and demonstrate how reconnection can catalyze large-scale recolonization in historically fragmented river networks.

California, Oregon↗

USGS2026

Bayesian belief network model to predict human-wildlife conflict in protected areas

Human-wildlife conflict (HWC) poses a pervasive global challenge, affecting livelihoods and threatening biodiversity. To better anticipate and mitigate HWC risk, we developed a large-scale predictive model using a Bayesian Belief Network (BBN). We surveyed 1,011 park rangers across 135 terrestrial protected areas in three Andean countries, documenting recent HWC incidents involving wildlife persecution or killing, livestock depredation, crop damage, or threats to human safety and property. We identified key drivers of HWC risk, including governance, wildlife acceptance, participation, and habitat quality. A sensitivity analysis revealed that enhancing governance and improving wildlife acceptance could reduce HWC risk by > 85%. The BBN model demonstrated scalability, effectively identifying strategies to reduce HWC risk at multiple scales, from individual protected areas to national networks. Our findings highlight the importance of strengthening governance, increasing wildlife acceptance, and enhancing community participation in conservation efforts. BBNs provide a flexible, cost-effective, and data-driven tool to guide protected areas and wildlife managers in monitoring, anticipating, and making informed decisions to mitigate conflict and promote coexistence.

Scientific Reports↗

USGS2026

Influence of new residential construction varying in housing density on bird species, human tolerance guilds, and communities

Human population growth and changing settlement patterns fuel the development of urban fringe lands worldwide, with implications for biodiversity. We conducted a 12-year study of birds in the fast-developing urban fringe lands of the central Puget Sound region, Washington, USA, to examine the effect of development configuration on birds. We hypothesized that lower-intensity conservation developments, compared to higher-intensity planned community developments, would benefit the overall bird community, as well as native forest birds (avoiders of human development) and avian generalist species (adapters to human development), but that higher-intensity planned community developments would benefit synanthropic species (exploiters of human development). We fit single-species and multi-species occupancy models to test these hypotheses. Consistent with our hypotheses, we found that a greater proportion of the overall community, avoiders, and adapters occupied lower-intensity conservation developments compared to higher-intensity planned community developments. However, we did not detect an effect of development type on the exploiter guild, and we found that species in the exploiter guild are variable in their response to the configuration of suburban developments. We also hypothesized that human tolerance guilds would be a useful predictor of individual species responses to development type. This hypothesis was somewhat supported: we found that, for avoiders, 87% of species in the guild had the same response to development type as the overall guild; for adapters, 63% had the same response as the overall guild, and for exploiters, only 44% had the same response as the overall guild. Our results indicate that the configuration of suburban developments can have a meaningful impact on bird communities, particularly on those species that are most sensitive to any level of development.

Washington↗