Real‐time PCR assay to detect brown marmorated stink bug, <i>Halyomorpha halys</i> (Stål), in environmental DNA

Valentin, Rafael; Maslo, Brooke; Lockwood, Julie L.; Pote, John; Fonseca, Dina M.

doi:10.1002/ps.4217

Cited by 25 publications

(31 citation statements)

References 46 publications

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“…To detect traces of BMSB DNA among the DNA extracted from the guano we employed a high-sensitivity real-time PCR assay specific to BMSB [30]. Briefly, we performed real-time PCR reactions in replicates of two using HPCL purified primers for targeting a conserved region of the rDNA internal transcribed spacer 1 (ITS1).…”

Section: Methodsmentioning

confidence: 99%

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Chirosurveillance: The use of native bats to detect invasive agricultural pests

et al. 2017

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Invasive insect pests cost the agricultural industry billions of dollars annually in crop losses. Timely detection of pests is critical for management efficiency. Innovative pest detection strategies, such as environmental DNA (eDNA) techniques, combined with efficient predators, maximize sampling resolution across space and time and may improve surveillance. We tested the hypothesis that temperate insectivorous bats can be important sentinels of agricultural insect pest surveillance. Specifically, we used a new high-sensitivity molecular assay for invasive brown marmorated stink bugs (Halyomorpha halys) to examine the extent to which big brown bats (Eptesicus fuscus) detect agricultural pests in the landscape. We documented consistent seasonal predation of stink bugs by big brown bats. Importantly, bats detected brown marmorated stink bugs 3–4 weeks earlier than the current standard monitoring tool, blacklight traps, across all sites. We highlight here the previously unrecognized potential ecosystem service of bats as agents of pest surveillance (or chirosurveillance). Additional studies examining interactions between other bat and insect pest species, coupled with comparisons of detectability among various conventional monitoring methods, are needed to verify the patterns extracted from this study. Ultimately, robust economic analyses will be needed to assess the cost-effectiveness of chirosurveillance as a standard strategy for integrated pest management.

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Section: Methodsmentioning

confidence: 99%

“…Full validation details of the BMSB assay are described in Valentin et al . [30]. We ran reactions at an initial denaturing step of 96°C for 10 minutes, followed by 45 cycles of denaturing at 96°C for 15 seconds and annealing and extension at 60°C for 1 minute.…”

Section: Methodsmentioning

confidence: 99%

Chirosurveillance: The use of native bats to detect invasive agricultural pests

et al. 2017

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“…To assess the presence of BMSB DNA in these samples, we used a TaqMan quantitative polymerase chain reaction (qPCR) assay designed specifically for BMSB (for details regarding assessments of specificity and sensitivity, see Valentin et al . ). Briefly, we used 20‐μl reactions with 500 nanomolar (nM) concentration of each primer, 250 nM of the probe, 1X TaqMan ® Environmental Master Mix 2.0, and 2 μl of DNA, following a reaction protocol with an initial denaturing step of 96°C for 10 min, followed by 45 cycles of denaturing for 15 s and annealing and extension at 60°C for 1 min.…”

Section: Methodsmentioning

confidence: 97%

“…We used a genetic tool we had previously designed for BMSB; this tool is very sensitive to trace amounts of degraded DNA and exclusively targets BMSB (Valentin et al . ; Maslo et al . ).…”

Section: Methodsmentioning

confidence: 99%

“…We used a genetic tool we had previously designed for BMSB; this tool is very sensitive to trace amounts of degraded DNA and exclusively targets BMSB (Valentin et al 2016;Maslo et al 2017). Because they are a sap-feeding species, BMSB individuals remain on a host plant for extended periods of time (Leskey et al 2012a), potentially leaving detectable amounts of DNA as they feed, defecate, and/or molt.…”

Section: Target Edna Collectionmentioning

confidence: 99%

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Early detection of invasive exotic insect infestations using eDNA from crop surfaces

Valentin

Fonseca

Nielsen

et al. 2018

Frontiers in Ecol & Environ

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The number of exotic species invasions has increased over recent decades, as have the ecological harm and economic burdens they impose. Rapid‐response eradication of nascent exotic populations is a viable approach to minimizing damage, but implementation is limited by the difficulty of detecting such species during the early stages of infestation due to their small numbers. The use of environmental DNA (eDNA) has helped address this issue in aquatic ecosystems, but to the best of our knowledge has not been trialed for surveillance of exotic species in terrestrial systems. Using a high‐resolution, real‐time (quantitative) polymerase chain reaction assay, we developed a highly efficient protocol to survey agricultural fields for the invasive non‐native brown marmorated stink bug (BMSB; Halyomorpha halys). We compared results using eDNA to those for conventional monitoring traps and documented substantially higher sensitivity and detection effectiveness. Our methodology is transferable to situations in which the DNA of terrestrial target species can be accumulated into a single substrate, suggesting that eDNA‐based approaches could transform our ability to detect exotic insects in non‐aquatic settings.

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The state, transport, and fate of aboveground terrestrial arthropod eDNA

et al. 2021

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Environmental DNA (eDNA) analyses have become invaluable for detecting and monitoring aquatic and terrestrial species and assessing site biodiversity within aquatic environments or soil. Recent studies have extended these techniques by using eDNA to identify the presence of aboveground terrestrial arthropods directly from vegetative surfaces. However, while the dynamics of eDNA state, transport, and fate (its “ecology”) have been explored within aquatic environments and soil, they have yet to be explored within aboveground terrestrial systems. Here, we explore the ecology of terrestrial eDNA deposited by fluid‐feeding arthropods on leaf surfaces. We carried out a series of experiments to evaluate the optimal filter pore size for intracellular eDNA collection, how eDNA is affected by rain events, and its degradation rate under different solar radiation conditions. We found that the captured concentration of intracellular eDNA was not significantly affected by an increase in filter pore size, suggesting a wide range of viable pore size options exist for targeting intracellular eDNA. We also found extracellular eDNA from fluid excrement degrades more rapidly than intracellular when exposed to solar radiation, indicating the latter is a more viable target for collection. Finally, we identified that rainfall or mist will remove most terrestrial eDNA present on vegetation surfaces. We provide researchers and environmental managers key insights into successfully designing and carrying out aboveground terrestrial arthropod eDNA surveys that maximize detection probability.

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Real‐time PCR assay to detect brown marmorated stink bug, Halyomorpha halys (Stål), in environmental DNA

Abstract: Given the high sensitivity of our assay to BMSB environmental DNA (eDNA) in terrestrial samples, this tool should become a cost-effective approach for using eDNA to detect terrestrial invasive species and their key predators. © 2016 Society of Chemical Industry.

Cited by 25 publications

References 46 publications

Chirosurveillance: The use of native bats to detect invasive agricultural pests

Chirosurveillance: The use of native bats to detect invasive agricultural pests

Early detection of invasive exotic insect infestations using eDNA from crop surfaces

The state, transport, and fate of aboveground terrestrial arthropod eDNA

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