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Cyanobacterial blooms in aquatic environments have impacted ecosystem health, altered food webs and contributed to substantial regional economic losses. The relative impacts of climate change, eutrophication and other environmental stressors on the formation of cyanobacterial blooms remain unclear as a consequence of the lack of long‐term data. The analysis of lake sediment archives can help address such questions.The abundance of target cyanobacterial genes was quantified using droplet digital PCR from two experimental and two reference lakes at the IISD ‐ Experimental Lakes Area, Northwestern Ontario, Canada where various anthropogenic drivers have been manipulated and phytoplankton biomass and taxonomy (via microscopy) tracked over the past 50 years.Based on sediment DNA (sedDNA) records, Lake 227 has seen more than two full orders of magnitude rise in cyanobacterial abundance in response to experimental nutrient (nitrogen and phosphorus) loading since 1969. Lake 223 also experienced a small increase in cyanobacterial abundance during a period of experimental acidification. The sedDNA archives also identified an increase in abundance of mcyE, a gene involved in microcystin synthesis, in Lake 227 after 1990 when experimental nitrogen loading ceased and only phosphorus loading continued. Sequencing of the cyanobacterial 16S rRNA gene from sedDNA revealed a shift towards Nostocales dominance in response to eutrophication in Lake 227; this was also seen in the more recent sediments of the other lakes but to a much lesser extent.When comparing the sedDNA data with historical phytoplankton records of the past ~50 years, moderate‐to‐strong correlations were found between the two. This research validated the use of sedDNA for the analysis of long‐term trends in lakes and the results revealed additional changes that were not recorded from the surface‐water records.Experimental nutrient loading had the greatest influence on the cyanobacterial community in Lake 227. However, climate change, via declines in heating degree days, was also significant in explaining variation in the cyanobacterial communities in the experimental lakes as well as the reference lakes. [ABSTRACT FROM AUTHOR]

The only population of the endangered blue racer (Coluber constrictor foxii) in Canada occurs on Pelee Island, Ontario. The species is threatened by multiple factors, including habitat degradation and loss, road mortality, persecution, and potentially predation. We designed and evaluated the performance of an environmental DNA droplet digital PCR assay that can be used for multiple facets of conservation of this species. We tested the assay in silico and in vitro using DNA of blue racers and co-occurring snake species and estimated the LOD and LOQ using synthetic DNA. As wild turkey predation has been suggested to negatively affect racers, we tested the assay on eight wild turkey faecal samples. Our assay is specific, can detect the target species at very low levels of concentration (0.002 copies/µL), and can accurately quantify copy numbers ≥ 0.26 copies/µL. We detected no racer DNA in any wild turkey faecal sample. More faecal samples collected at strategic locations during snake peak activity on Pelee Island would enable a more thorough assessment of the possibility of turkey predation. Our assay should be effective for other environmental samples and can be used for investigating other factors negatively affecting blue racers, for example, helping to quantify blue racer habitat suitability and site occupancy. [ABSTRACT FROM AUTHOR]

Background: Climate change has driven shifts in breeding phenology of many amphibians, causing phenological mismatches (e.g., predator-prey interactions), and potentially population declines. Collecting data with high spatiotemporal sensitivity on hibernation emergence and breeding times can inform conservation best practices. However, monitoring the phenology of amphibians can be challenging because of their cryptic nature over much of their life cycle. Moreover, most salamanders and caecilians do not produce conspicuous breeding calls like frogs and toads do, presenting additional monitoring challenges. Methods: In this study, we designed and evaluated the performance of an environmental DNA (eDNA) droplet digital PCR (ddPCR) assay as a non-invasive tool to assess the breeding phenology of a Western Chorus Frog population (Pseudacris maculata mitotype) in Eastern Ontario and compared eDNA detection patterns to hourly automatic acoustic monitoring. For two eDNA samples with strong PCR inhibition, we tested three methods to diminish the effect of inhibitors: diluting eDNA samples, adding bovine serum albumin to PCR reactions, and purifying eDNA using a commercial clean-up kit. Results: We recorded the first male calling when the focal marsh was still largely frozen. Chorus frog eDNA was detected on April 6th, 6 days after acoustic monitoring revealed this first calling male, but only 2 days after males attained higher chorus activity. eDNA signals were detected at more sampling locales within the marsh and eDNA concentrations increased as more males participated in the chorus, suggesting that eDNA may be a reasonable proxy for calling assemblage size. Internal positive control revealed strong inhibition in some samples, limiting detection probability and quantification accuracy in ddPCR. We found diluting samples was the most effective in reducing inhibition and improving eDNA quantification. Conclusions: Altogether, our results showed that eDNA ddPCR signals lagged behind male chorusing by a few days; thus, acoustic monitoring is preferable if the desire is to document the onset of male chorusing. However, eDNA may be an effective, non-invasive monitoring tool for amphibians that do not call and may provide a useful complement to automated acoustic recording. We found inhibition patterns were heterogeneous across time and space and we demonstrate that an internal positive control should always be included to assess inhibition for eDNA ddPCR signal interpretations. [ABSTRACT FROM AUTHOR]