Use of qPCR-Based Cercariometry to Assess Swimmer’s Itch in Recreational Lakes

Rudko, Sydney P.; Reimink, Ronald L.; Froelich, Kelsey; Gordy, Michelle A.; Blankespoor, Curtis L.; Hanington, Patrick C.

doi:10.1007/s10393-018-1362-1

Cited by 31 publications

(74 citation statements)

References 47 publications

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“…It is possible that the use of pre-filtration steps may be required to exclude whole organisms, potentially providing more standardised quantitative spatial and temporal comparisons [33]. Alternatively, it would be possible to modify the environmental DNA methods used here to focus sampling exclusively cercariae [46,47]. Such "qPCR cercariometry" requires relatively large volumes of water (e.g.…”

Section: Plos Neglected Tropical Diseasesmentioning

confidence: 99%

“…Such "qPCR cercariometry" requires relatively large volumes of water (e.g. 25 litres) that are filtered through a 20μm mesh zooplankton net, before residual material is collected and concentrated allowing qPCR analyses [46,47]. This cercariometry method would allow the collection of large volumes of genetic material from the environment; however such zooplankton nets are expensive and their re-use requires decontamination between sampling events if they are to remain effective.…”

Section: Plos Neglected Tropical Diseasesmentioning

confidence: 99%

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Schistosoma species detection by environmental DNA assays in African freshwaters

et al. 2020

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Background Schistosomiasis is a neglected tropical parasitic disease associated with severe pathology, mortality and economic loss worldwide. Programs for disease control may benefit from specific and sensitive diagnostic methods to detect Schistosoma trematodes in aquatic environments. Here we report the development of novel environmental DNA (eDNA) qPCR assays for the presence of the human-infecting species Schistosoma mansoni, S. haematobium and S. japonicum. Methodology/Principal findings We first tested the specificity of the assays across the three species using genomic DNA preparations which showed successful amplification of target sequences with no cross amplification between the three focal species. In addition, we evaluated the specificity of the assays using synthetic DNA of multiple Schistosoma species, and demonstrated a high overall specificity; however, S. japonicum and S. haematobium assays showed cross-species amplification with very closely-related species. We next tested the effectiveness of the S. mansoni assay using eDNA samples from aquaria containing infected host gastropods, with the target species revealed as present in all infected aquaria. Finally, we evaluated the effectiveness of the S. mansoni and S. haematobium assays using eDNA samples from eight discrete natural freshwater sites in Tanzania, and demonstrated strong correspondence between infection status established using eDNA and conventional assays of parasite prevalence in host snails. Conclusions/Significance Collectively, our results suggest that eDNA monitoring is able to detect schistosomes in freshwater bodies, but refinement of the field sampling, storage and assay methods are PLOS NEGLECTED TROPICAL DISEASES

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Section: Plos Neglected Tropical Diseasesmentioning

confidence: 99%

Section: Plos Neglected Tropical Diseasesmentioning

confidence: 99%

Schistosoma species detection by environmental DNA assays in African freshwaters

et al. 2020

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“…The 18S avian schistosomes-targeting qPCR assay was performed as described in Rudko et al (2018) [21] (S1 Table). The LOD 95 of this technique is 3.4 gene copies/ rxn [21]. qPCR master mix (IDT DNA) containing 1x master mix, and 200nm forward reverse primer and a fluorescein-labelled probe were used.…”

Section: Qpcr Methodsmentioning

confidence: 99%

“…Avian schistosome monitoring. Sample collection was conducted as described in Rudko et al (2018). Briefly, 25L water samples, collected one litre at a time across a shoreline up to…”

Section: Sample Collectionmentioning

confidence: 99%

“…In 2018, the avian schistosomes monitoring group was interested in transitioning to a DNA extraction method that would allow for batch processing of samples. We, therefore, opted to transition their program to the DNeasy DNA extraction kit (Rudko et al 2018). To set up this remote laboratory in a cost-effective manner, equipment (centrifuge, heating block, and vortex) were sourced from Dot Scientific (USA) (S2 Table), and pipettes were from the company VWR (USA).…”

Section: Dna Extractionmentioning

confidence: 99%

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Democratizing water monitoring: Implementation of a community-based qPCR monitoring program for recreational water hazards

Rudko

Reimink

Peter³

et al. 2020

PLoS ONE

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Recreational water monitoring can be challenging due to the highly variable nature of pathogens and indicator concentrations, the myriad of potential biological hazards to measure for, and numerous access points, both official and unofficial, that are used for recreation. The aim of this study was to develop, deploy, and assess the effectiveness of a quantitative polymerase chain reaction (qPCR) community-based monitoring (CBM) program for the assessment of bacterial and parasitic hazards in recreational water. This study developed methodologies for performing qPCR 'in the field,' then engaged with water management and monitoring groups and tested the method in a real-world implementation study to evaluate the accuracy of CBM using qPCR both quantitatively and qualitatively. This study found high reproducibility between qPCR results performed by non-expert field users and expert laboratory results, suggesting that qPCR as a methodology could be amenable to a CBM program.

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Efficacy of environmental DNA sampling to detect the occurrence of endangered coho salmon (Oncorhynchus kisutch) in Mediterranean‐climate streams of California's central coast

et al. 2020

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Analysis of environmental DNA (eDNA) has emerged as an important tool for investigating the occurrence and distribution of fish and other species in aquatic environments. However, in lotic systems, uncertainty remains about how environmental factors influence the downstream transport, degradation, and dilution of eDNA, and hence how detection probability varies with distance from the eDNA source. We conducted cage experiments and paired eDNA and snorkel surveys to evaluate the potential for eDNA methods to detect endangered coho salmon in Mediterraneanclimate streams of the Santa Cruz Mountains, California. Juvenile coho salmon at two densities were placed in cages, and water samples were collected and analyzed (qPCR) at seven locations from 10 to 1,000 m downstream. Although we detected coho salmon eDNA up to 1,000 m downstream of the cage, the vast majority of detections were within 200 m, and detections at greater distances occurred only at the higher fish density. In field collections from 21 stream reaches across the Santa Cruz Mountains, eDNA methods and snorkeling produced identical outcomes in terms of reach-level detection of coho salmon. Probability of occurrence in a water sample and detection in a qPCR replicate both increased with observed fish density; however, even at the lowest densities, a protocol of three water samples and two qPCR replicates resulted in a >90% cumulative probability of detection. Overall, our studies suggest that detection probability decreased rapidly with distance in Santa Cruz Mountain streams, likely because low stream discharges, slow transport velocities, and strong interaction between the water column and stream substrate result in rapid degradation or settling of eDNA. Our findings thus support the use of eDNA methods for detecting rare species, but also indicate that local conditions strongly influence transport dynamics and hence need to be considered when developing survey designs.

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Use of qPCR-Based Cercariometry to Assess Swimmer’s Itch in Recreational Lakes

Cited by 31 publications

References 47 publications

Schistosoma species detection by environmental DNA assays in African freshwaters

Schistosoma species detection by environmental DNA assays in African freshwaters

Democratizing water monitoring: Implementation of a community-based qPCR monitoring program for recreational water hazards

Efficacy of environmental DNA sampling to detect the occurrence of endangered coho salmon (Oncorhynchus kisutch) in Mediterranean‐climate streams of California's central coast

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